page 5 - 17th Meeting of the European Association for

Transcription

Turkish Society of Hematology
17
th Meeting of the
European Association for
Haematopathology
PROGRAM AND ABSTRACT BOOK
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PROGRAM
and
ABSTRACT BOOK
17th Meeting of the
European Association for Haematopathology
17-22 October 2014
Hilton İstanbul Bosphorus Hotel
İstanbul - TURKEY
www.eahp2014.org
WELCOME MESSAGES
Isinsu Kuzu, Chair of the Local Organizing Committee
Elias Campo, President of the EAHP
7
CONGRESS COMMITTEES
10
CONGRESS VENUE
11
GENERAL INFORMATION
12
LOCATION OF ACTIVITIES
14
TOPIC & MAIN GOAL
16
SCIENTIFIC PROGRAM
17
SATELLITE SYMPOSIUMS
32
BIOSKETCH & PHOTO OF INVITED SPEAKERS &
ORAL PRESENTERS
35
ORAL PRESENTATIONS
51
POSTER PRESENTATIONS
69
SOCIAL PROGRAM
133
İstanbul RAIL LINES NETWORK MAP
138
TOURIST INFORMATION
139
TURKEY
143
OFFICIAL CARRIER
144
TOURS
İstanbul Tours Information
Pre & Post Congress Tours Information
145
SUPPORTING INSTITUTIONS & EXHIBITORS
151
EXHIBITION FLOOR PLAN
152
CORRESPONDENCE
153
INDEX OF ABSTRACT AUTHORS
155
İ S TA N B U L - T U R K E Y
| 17-22 October 2014
| EAHP - 2014
|
C ON T EN T S
17 th MEETING OF THE EUROPEAN ASSOCIATION FOR HAEMATOPATHOLOGY
CONTENTS
5
On behalf of the Local Organizing Committee it is my pleasure and honour to welcome you to the 17th EAHP meeting organized in the magnificent city of İstanbul.
The EAHP Executive Committee and the Bone Marrow Working Group, with the
support of distinguished scientists, have organized the scientific program to integrate basic scientific knowledge, diagnostic and clinical aspects of the small B cell
lymphomas.
The educational session will cover the principals of new technologies and their transfer to the clinical
practice, which is an important challenge for the future.
The Bone Marrow Symposium and Workshop will cover CLL and bone marrow involvement by other
small B cell lymphomas. In the Symposium 21 scientific studies will be discussed either as a platform
or poster presentation. There are 88 interesting cases accepted for the Bone Marrow Workshop
which will be discussed in three sessions.
The Lymphoma Symposium includes lectures by distinguished guest speakers as well as platform
presentations of 24 scientific studies selected from 155 submitted abstracts. The remaining scientific
studies will be presented as posters and discussed in the poster session. In addition there will be
three satellite symposia supported by industry during the Lymphoma Symposium.
We received 193 very interesting cases submitted for the Lymphoma Workshop. These cases will
be discussed in 6 sessions and will be presented either by the submitters or by the lymphoma panel
members.
W EL C OM E M ES S A GES
Dear Participants
WE LC O M E WO RD S O F THE CHAIR
The digital images of the original H&E stained slides of Bone Marrow and Lymphoma Workshops
cases together with their corresponding clinical, pathological and molecular annotations are now
available to access on the web site for each participant.
While we appreciate that the meeting program is busy we hope that you will relax at the social events
of the program. The Congress venue and the Taksim area, which is at the heart of the city is convenient for you to reach many places. We would recommend that you try to leave some time to explore
some of the spectacular parts of the unique city of İstanbul.
I wish you a scientifically and socially memorable meeting in İstanbul.
Isinsu Kuzu
Chair of the Organizing Committee
| 17-22 October 2014
| EAHP - 2014
|
7
It is my privilege to welcome you all to İstanbul on behalf of the European Association for Haematopathology (EAHP) ExCo members and Dr Isinsu Kuzu, as Chair of
the enthusiastic and dedicated Local Organizing Committee. Working together, the
two committees have prepared an excellent program for the XVII EAHP congress
that I hope will fulfill your expectations. These meetings are a partnership between
our Association and the Society for Hematopathology. The increasing success of
our congresses is a natural consequence of the intense and fruitful scientific collaboration of our members that is now expanding to colleagues from all around the world. As you
know these meetings move around the European geography, most recently from South (Bordeaux),
North (Uppsala), West (Lisbon) and now to the East in the magnificent city of İstanbul, bridge of continents, history and cultures. The move of our meetings around Europe and the growing international
participation are a reflection of the growing scientific networks and collegiality of all the participants
with the common goal of promoting knowledge and education in Haematopathology.
As our deeply missed friend David Y Mason used to say, each technological advance applied to our
Pathology work opens a window onto a new landscape of knowledge. We are on the verge of a new
step forward in Pathology with the progress triggered by the new DNA sequencing and other technologies that run in parallel with the development of new targeted drugs and management strategies
for our patients. All this new knowledge has to be integrated with our solid experience from classical
pathology. This challenge is what the exciting program of this meeting on “Redefining the spectrum of
small B-cell lymphomas in light of current technology” will try to accomplish.
W EL C OM E M ES S A GES
Dear colleagues
WE LC O M E WO RD S O F THE PRESIDENT
I would like to highlight our 2014 Karl Lennert awardee Dr. Nancy Lee Harris. This EAHP recognition
honors an outstanding haematopathologist for their inspiring contributions that have been instrumental in reshaping the understanding of our specialty. It is also with great satisfaction that I would like to
introduce the David Y Mason Award, a new feature of this meeting thanks to the generous contribution of the David Y Mason Foundation. This Award aims to foster the interest of young researchers for
Haematopathology and help them to become active members of our Society. Thanks to the generosity of our members we have continued and expanded our EAHP Travel Grant Program to facilitate the
participation of young haematopathologists and scientists from developing countries to attend our
meetings. We hope this initiative will continue to increase in the future.
Although I am sure the Scientific Program will keep you very busy here in the congress venue, do not
forget to devote time to experience the fascinating city of İstanbul. Few places in the world can offer
such an amalgam of history and culture.
Finally, I would like to deeply thank all the sponsors for helping us make this event possible, all EXCO,
Panel and European Bone Marrow Group members for their great contribution to the organization of
the meeting with my special recognition to Dr. Isinsu Kuzu, Leticia Quintanilla-Fend, Daphne de Jong,
Andrew Wotherspoon, and Philippe Gaulard who had been continuously dedicated to making this
meeting an unforgettable experience.
On behalf of the EXCO members and of Dr. Isinsu Kuzu, welcome to İstanbul!!
Elias Campo
President of the EAHP
| 17-22 October 2014
| EAHP - 2014
|
9
17 MEETING OF THE EUROPEAN ASSOCIATION FOR HAEMATOPATHOLOGY
ORGANIZING COMMITTEE
INTERNATIONAL SCIENTIFIC COMMITTEE
Chair: Isinsu Kuzu
Elias Campo (chair)
Pierre Brousset
Jose Cabecadas
LOCAL ORGANIZING COMMITTEE
Leticia Quintanilla Fend
Nalan Akyurek
Philippe Gaulard
Oner Dogan
John Goodlad
Mine Hekimgil
Daphne de Jong
Mükerrem Safali
Marsha Kinney
Nukhet Tuzuner
Isinsu Kuzu
Aysegul Uner
Attilio Orazi
Suheyla Uyar
German Ott
Elena Sabattini
Birgitta Sander
Andrew Wotherspoon
th
C ON GRES S C OM M I T T EES
C O NG RE SS C OMMI TTEES
BONE MARROW WORKSHOP PANEL MEMBERS
LYMPHOMA WORKSHOP PANEL MEMBERS
Attilio Orazi
Elias Campo
Falko Fend
John Chan
Marcus Kremer
Stephan Dirnhofer
Anna Porwit
Ahmet Dogan
Mukerrem Safali
Leticia Quintanilla-Fend
Jon Van der Walt
Isinsu Kuzu
German Ott
Birgitta Sander
Steven H. Swerdlow
Luc Xerri
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| EAHP - 2014
| 17-22 October 2014
| İ S TA N B U L - T U R K E Y
İstanbul Hilton & Convention Center
Address
Phone
Fax
Web
: Cumhuriyet Cad. Harbiye 34367 İstanbul, Turkey
: +90 212 315 60 00
: +90 212 240 41 65
: www.hiltonistanbul.com.tr
C ON GRES S VEN UE
C O N G R E SS VE NUE
Surrounded by 15 acres of beautiful gardens, Hilton İstanbul Bosphorus stands in the very heart of the
city overlooking the Bosphorus Strait. Just minutes away from the city’s business, shopping and entertainment districts such as Taksim and Nisantasi, the hotel is also conveniently located next to Lutfi
Kirdar and İstanbul Congress Centre.
Hilton İstanbul Bosphorus is located just 50 minutes from Atatürk International Airport and 80 minutes
from Sabiha Gökçen Airport. The hotel is within easy walking distance of the famous Taksim leisuredistrict, exclusive Nișantașı shopping area and İstanbul Congress Centre
Airport Transportation
Airport Name
Atatürk Airport
Sabiha Gökçen Airport
Distance
35 km
60 km
Duration
50 minutes
80 minutes
| 17-22 October 2014
| EAHP - 2014
|
11
GEN ERA L I N FORM AT I ON
th
G E NE R A L I NFORMATI ON
Registration and Information Desk
Registration and information desks for the EAHP 2014 İstanbul will be held at the foyer of the
Hilton İstanbul Convention Center during the following hours:
October 17, 2014
07:30-18:30
October 18, 2014
07:30-18:30
October 19, 2014
07:30-18:30
October 20, 2014
07:30-18:30
October 21, 2014
07:30-18:30
October 22, 2014
08:00-17:00
Official Language
Official language of the meeting and of correspondence is English.
There will be no simultaneous translation.
Name Badges
Please wear your name badges at all times during the Meeting.
Badges are color coded as follows:
Speaker, Organizing Committee Members
Red
Pack - Members, Non Members, Residents
Yellow
Lymphoma Symposium - Members, Non Members, Residents
Light Blue
Lymphoma Workshop - Members, Non Members, Residents
Green
Bone Marrow - Members, Non Members, Residents
Orange
Educational Session Members, Non Members, Residents
White
Industry
Purple
Accompany Person
Pink
Serenas Tourism (Organizing Secretariat)
Dark Blue
Speakers’ Room
Speakers should hand in presentations in the slide check and speakers’ room, located balcony in the main meeting
hall at the Hilton Convention Center as soon as possible after their arrival. Speakers must ensure that all files
needed for the presentation are included in the media of their choice (CD, USB Device) and they should be tested
on a computer other than that on which it was created. Prior to the scientific session, the authors should review their
presentations to ensure that it transferred properly.
CME Accreditation
17th Meeting of the European Association for Haematopathology is granted 30 European CME credits (ECMEC) by
the European Accreditation Council for Continuing Medical Education (EACCME). The CME forms will be given at
the end of the congress from the information kiosks
The EACCME credit system is based on 1 ECMEC per hour with a maximum of 3 ECMECs for half a day and 6
ECMECs for a full-day event.
12
| EAHP - 2014
| 17-22 October 2014
: 2 ECMECs
 18th October: Bone Marrow Symposium
: 3 ECMECs and Bone Marrow Workshop : 3 ECMECs
 19th and 20th October: Lymphoma Symposium
: 5 ECMECs for each day;
 21st and 22nd October: Lymphoma Workshop
: 6 ECMECs for each day.
European “CME” credits could be converted to the US CME Credits.
Please visit: http://www.ama-assn.org/ama/pub/education-careers/continuing-medical-education/physiciansrecognition-award-credit-system/other-ways-earn-ama-pra-category/international-programs/uemseaccme-creditconversion.page
Certificate of Attendance
It will be given at the end of the congress from the Registration and Information Desks.
Internet Access
Internet cafes are available all around the city and wireless internet access for notebook users at the Hilton
Convention Center will be provided by the organizational secretariat Serenas Tourism.
Cloakroom
The cloakroom is in the ground floor of Hilton Convention Center.
Mobile Phones
Delegates are kindly asked to verify that mobile phones are switched off (silent mode) during sessions, as a courtesy
for speakers and attendees.
GEN ERA L I N FORM AT I ON
The 17th EAHP Congress was accredited as follows:
 17th October: Educational Session
Coffee Breaks
Coffee breaks will be served in exhibition area at the coffee break times. The coffee tickets are inserted in to the
badges and will be asked during the service.
Lunch
Lunch will be served as openbuffet at the exhibition area on October 18–21 and 22. October 19th – 20th days will
be served as lunchbox at the main meeting hall during the satellite symposium. Lunch tickets are inserted into the
badges and will be asked during the service.
Posters
The posters will be displayed between 17–22 October, 2014 at the Poster Area of the Exhibition Hall according to
the following schedule:
Poster Mount Date & Time
Poster Remove Date & Time
Discussion Date & Time
: October 17, 2014 at 14:00
: October 22, 2014 at 14:00
: October 20, 2014 at 14:15 – 15:30
Bone Marrow Poster Presentations
Lymphoma Poster Presentations
TOTAL
: 17 posters
:114 posters
:131 Posters
PP-BM-01 to PP-BM-17
PP-LYMP-001 to PP-LYMP-114
The organization secretariat is not responsible for the posters that haven’t been removed after the session.
Poster discussions will take place in front of the posters between 14:15-15:30.
Presenters should be beside their posters during the discussion dates and times.
Meet the Professor Session
Meet the Professor Session tickets will be collected from the registration desk during the meeting.
The total capacity for each session is 40 pax.
| 17-22 October 2014
| EAHP - 2014
|
13
ACTIVITY
LOCATION
FLOOR
Registration Desk
Hilton Convention Upper Hall Foyer
Main Meeting Hall
Hilton Convention Upper Hall
Speakers’ Room
Hilton Convention Upper Hall Balcony
LYWS Slide Box Collection
Hilton Convention Upper Hall Balcony
Exhibition Area
Hilton Convention Lower Hall
Poster Area
Hilton Convention Lower Hall
Storage Room
Hilton Convention Lower Hall Foyer
Meet the Professor Sessions / Mercury Room
Hilton Hotel
Lobby Floor
Meet the Professor Sessions / Lalezar Room
Hilton Hotel
Lobby Floor
Opening Cocktail / Ball Room
Hilton Hotel
-1 Floor
th
L OC AT I ON OF A C T I VI T I ES
LO C AT IO N OF A CTI VI TI ES
LYWS SLIDE BOX COLLECTION
(Balcony)
SPEAKER &
SOCIAL PROGRAM
DESK
Entrance of Main Meeting Hall
Lower Hall
Exhibition Area / Poster Area / Coffee and Lunch Area / Storage Room
14
| EAHP - 2014
| 17-22 October 2014
MAIN ENTRANCE
REGISTERED
PARTICIPANT
ONSITE / CASHIER
DESK
SPEAKERS’ ROOM
(Balcony)
COFFEE BREAK AREA
EXHIBITION AREA
STAGE
MAIN MEETING HALL
| 17-22 October 2014
| EAHP - 2014
|
? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?L?OC
???
AT
? ?I ON
? ? ?????????????
OF A C T I VI T I ES
CONVENTION
ENTRANCE
POSTER AREA
SPEAKERS’ ROOM
(Balcony)
EXHIBITION AREA
EXHIBITION AREA
STORAGE
ROOM
FOYER
15
Bone Marrow Symposium Topic
“Small B-cell lymphoid neoplasms in bone marrow: diagnostic advances”
Main Goal
The Bone Marrow Symposium will be focused on small B-cell lymphoid neoplasms, particularly those predominantly
studied and diagnosed in the bone marrow and peripheral blood. Particular relevance will be given to the use of
newer technologies which by more precisely defining these entities will provide effective guidance in therapeutic
decisions.
Lymphoma Symposium Topic
“Redefining the spectrum of small B-cell lymphomas in light of current technology”
New entities versus extremes of spectrum of well recognized categories
Molecular genetic mechanisms in the progression of small B-cell lymphomas
Clinical relevance
th
? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?T?OP
? ?I?C? ?
A?????????????
N D M A I N GOA L
T O P IC A ND MA I N GOA L
Main Goal
The Lymphoma Symposium will be devoted to new perspectives in the understanding of the spectrum of small B-cell
lymphoid neoplasms, particularly in light of the information provided by new technologies and tools in the study of
these tumors. We will address B-cell lymphoma categories or variants and explore how new phenotypic, molecular
and genetic information may help to better recognize these entities and help us to understand their different clinical
and pathological presentations. We will discuss recent technological advances and evaluate the clinical impact of
this new knowledge with particular emphasis in the potential value to guide diagnostic and therapeutic decisions.
SCIENTIFIC PROGRAM COLOUR LEGEND
ES
: Educational Session
BMS S
: Bone Marrow Symposium Session
BMW S
: Bone Marrow Workshop Session
LYMP S
: Lymphoma Symposium Session
LYMP W
: Lymphoma Workshop
MP
: Meet the Professor
PP S
: Proffered Papers Session
PS
: Poster Discussion
PS
: Poster Session
OC
: Social Activities
Karl Lennert Lecture
EBMWG Business Meeting
Satellite Symposium
16
| EAHP - 2014
| 17-22 October 2014
17 October 2014
Friday
18 October 2014
Saturday
PROGRAM OVERVIEW PER DAY
19 October 2014
Sunday
20 October 2014
Monday
21 October 2014
Tuesday
Meet the Professor
Session
Meet the Professor
Session
Meet the Professor
Session
Lymphoma Symposium
Session 3
Key Note Lecture 3
Lymphoma Symposium
Session 5
Topic Lecture 5
22 October 2014
Wednesday
07:15-07:30
07:30-08:00
08:00-08:15
08:15-08:30
Welcome Words
08:30-08:45
08:45-09:00
Bone Marrow
Symposium Session 1
09:00-09:15
09:15-09:30
EAHP Lymphoma
Symposium Session 1
Key Note Lecture 1
EAHP Lymphoma
Symposium Session 1
Topic Lecture 1
09:30-09:45
Coffee Break
10:30-10:45
Coffee Break
Coffee Break & Poster
View
10:45-11:00
11:00-11:15
Bone Marrow
Symposium Session 2
REGISTRATION
12:30-12:45
EAHP Lymphoma
Symposium 2
Topic Lecture 2
Proffered Papers
2a
Lunch
12:45-13:00
Coffee Break
Coffee Break & Poster
view
EAHP Lymphoma
Symposium 2
Key Note Lecture 2
11:15-11:30
12:15-12:30
Proffered Papers
Session 5
Proffered Papers
Session 1
10:00-10:30
11:45-12:00
12:00-12:15
Lymphoma Symposium
Session 3
Topic Lecture 3
Proffered Papers
Session 3
09:45-10:00
11:30-11:45
EAHP Lymphoma
Workshop Session 4
Lymphoma Symposium
Session 4
Key Note Lecture 4
EAHP Lymphoma
Workshop Session 5
Proffored Papers
Session 4
EAHP Lymphoma
Workshop Session 1
S C I EN T I F I C P ROGRA M M E
TIMING
S C IE N T I FI C P ROGRA M
Lunch Box Serve
Lunch Box Serve
Lunch & Satellite
Symposium
13:00-13:15
POSTER SESSION
13:15-13:30
13:30-13:45
13:45-14:00
Lunch & Satellite
Symposium
Roche
Lunch
Lunch
Poster Discussion
EAHP Lymphoma
Workshop Session 2
EAHP Lymphoma
Workshop Session 6
Coffee Break
Closing remarks
Janssen
14:00-14:15
Bone Marrow Workshop
Session 1
14:15-14:30
14:30-14:45
14:45-15:00
15:00-15:15
Coffee Break and
Poster Viewing
15:15-15:30
Proffered Papers 2b
15:30-15:45
Coffee Break
15:45-16:00
Karl Lennert Lecture
Session 2
16:00-16:15
16:15-16:30
16:30-16:45
Session 3
16:45-17:00
17:00-18:00
Satellite Symposium
EBMWG Business
Meeting
18:00-18:30
18:30-19:00
19:00-19:30
19:30-20:00
Coffee Break and
Poster Viewing
Educational Session
Opening Ceremony
DAVID Y MASON
LECTURE
Michael R Stratton, UK
GENERAL
ASSEMBLY
EAHP Lymphoma
Workshop Session 3
Takeda
Bus Transfer from
Hilton İstanbul to
Kuruçeşme Port
Gala Dinner
Bus Transfer from
Kuruçeşme Port to Hilton
İstanbul
| 17-22 October 2014
| EAHP - 2014
|
17
th
18
17 OCTOBER 2014, FRIDAY
18:00-20:00
EDUCATIONAL SESSION
Chairs: Adam Bagg, USA and Pierre Brousset, France
18:00
Rational use of genetic testing in B-cell lymphomas: where are we now?
Adam Bagg, Philadelphia, USA
18:30
FISH strategies and applications in lymphoma diagnosis
Jean Philippe Merlio, Bordeaux, France
19:00
Principles and approaches of high throughput sequencing
Xose Puente, Oviedo, Spain
19:30
High throughput sequencing in lymphoma: moving to the diagnostic arena
Frédéric Davi, Paris, France
20:00
Transcriptional analysis in lymphoma and translation into clinical practice
Lisa Rimsza, Tucson, AZ, USA
| EAHP - 2014
| 17-22 October 2014
MAIN HALL
08:30-09:00
09:00-09:30
BONE MARROW SYMPOSIUM SESSION 1
MAIN HALL
Chairs: Hans Kreipe, Carlos Bueso-Ramos
Understanding the biology CLL in the light of newer technologies
Richard Rosenquist, Uppsala, Sweden
Defining the genetic boundaries of other (non-CLL) small B cell lymphoid neoplasms
typically encountered in bone marrow:
Brunangelo Falini, Perugia, Italy
09:30-10:00
10:00-11:00
Coffee Break with Poster View
BONE MARROW SYMPOSIUM SESSION 2
MAIN HALL
10:00-10:30
Chairs: Hans Michael Kvasnicka, Bogna Wroblewska
Targeted therapy strategies for both CLL and other small B cell neoplasms:
10:30-11:00
Morphology and immunohistologic assessment of bone marrows involved by small B cell
lymphoid neoplasms.
Armando López-Guillermo, Barcelona, Spain
Elena Sabattini, Bologna, Italy
11:00-12:00
PROFFERED PAPERS
Chairs: Stephan Dirnhofer, Konnie Hebeda
11:00-11:15
08:30-09:30
18 OCTOBER 2014, SATURDAY
[OP-BM-01]
Complete response of ETP-LL to a Notch pathway inhibitor: Molecular and cellular
characterization of a sentinel case
Jon C. Aster1, Ami Bhatt2, Birgit Knoechel2, Daniel J. Deangelo2, Li Pan1, Michael J. Kluk1,
Frank Kuo1, Matthew Meyerson2
1
Department of Pathology, Brigham and Women’s Hospital, Boston, MA USA
2
Department of Medical Oncology, Dana Farber Cancer Institute, Boston, MA USA
11:15-11:30
[OP-BM-02]
Epigenetic profiling of CLL reveals novel DNA methylation-based clusters and novel
mechanisms of lymphomagenesis
Fang Fang1, Julia Geyer2, Wayne Tam2, Richard R. Furman1, Yi Fang Liu2, Peter Ouillette3,
Erlene Seymour3, Kamlai Saiya Cork3, Kerby Shedden4, Daniel M Knowles2, Ari Melnick1,
Sami N. Malek3, Attilio Orazi2, Rita Shaknovich2
1
Division of Hematology and Oncology, Weill Cornell Medical College, New York, USA
2
Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, USA
3
Department of Internal Medicine, Division of Hematology and Oncology, University of Michigan,
Ann Arbor, USA
4
Department of Statistics, University of Michigan, Ann Arbor, MI, USA
11:30-11:45
[OP-BM-03]
Non-IgM lymphoplasmacytic lymphoma shows pathologic and clinical heterogeneity and
may harbor MYD88 mutations
Rebecca L. King1, Wilson I. Gonsalves2, Matthew T. Howard1, Lori A. Frederick1,
David S. Viswanatha1, Patricia T. Greipp3, Stephen M. Ansell2, William G. Morice1
1
Division of Hematopathology, Mayo Clinic, Rochester, MN, USA
2
Department of Hematology/Oncology, Mayo Clinic, Rochester, MN, USA
3
Division of Laboratory Genetics, Mayo Clinic, Rochester, MN, USA
11:45-12:00
[OP-BM-04]
Hematopoietic stem cell origin of hairy cell leukemia
Stephen S Chung1, Eunhee Kim1, Jae H Park1, Young Rock Chung1, Julie Feldstein1,
Wenhuo Hu1, Michael F Berger1, Ari M Melnick2, Neal Rosen1, Martin S Tallman1,
Omar Abdel Wahab1, Christopher Y Park1
1
Memorial Sloan Kettering Cancer Center, New York, NY, USA
2
Weill Cornell Medical College, New York, NY, USA
12:00-13:00
Lunch
| 17-22 October 2014
| EAHP - 2014
|
19
POSTER SESSION
Chairs: Umberto Gianelli, Roos Leguit
14:00-15:00
BONE MARROW WORKSHOP SESSION 1: Diagnostic difficulties in CLL
Chairs: Jon van der Walt, Mukerrem Safali
Introduction
BMWS 55
Filiz Sen, New York, NY, USA
BMWS 253
Govind Bhagat, New York, NY, USA
BMWS 73
Gabriel Caponetti, Omaha, NE, USA
BMWS 139
Flavia Zacchi, Sao Paulo, Brazil
Summary and Conclusions
PANEL REVIEW
BMWS0064, BMWS0112, BMWS0138, BMWS0171, BMWS0172, BMWS0174, BMWS0191,
BMWS0309, BMWS0320
Coffee Break and Poster View
15:00-15:30
15:30-16:45 BONE MARROW WORKSHOP SESSION 2: Bone marrow manifestations of
LPL, MZL, SRPL, HCL-v and HCL
Chairs: Attilio Orazi, Marcus Kremer
Introduction
BMWS 83
Megan Nakashima, Cleveland, OH, USA
BMWS 194 John KarlFredericksen, Ann Arbor, MI, USA
BMWS 310 Mats Ehinger, Lund, Sweden
BMWS 121 Birgit Federmann, Tubingen, Germany
BMWS 195 Ling Zhang, Dallas, USA
PANEL REVIEW
BMWS0296, BMWS0306, BMWS0318
th
13:00-14:00
16:45-18:00 BONE MARROW WORKSHOP SESSION 3: Small B-cell lymphomas in bone
marrow multiclonality and composite lymphomas
Chairs: Anna Porwit, Falko Fend
Introduction
BMWS 289
Monika Klimkowska, Stockholm, Sweden
BMWS 300
Luis Colomo, Barcelona, Spain
BMWS 59
Duygu Kankaya, Ankara, Turkey
BMWS 160
Ulrika Klopcic, Ljubljana, Slovenia
BMWS 266
Xiangrong (Alex) Zhao, Bethesda, USA
PANEL REVIEW
BMWS0296, BMWS0306, BMWS0318
18:00-18:30
20
| EAHP - 2014
EBMWG Business Meeting
| 17-22 October 2014
MEET the PROFESSOR
MERCURY HALL
Daphne de Jong, Amsterdam, The Netherlands
“May be Hodgkin Lymphoma, maybe not”
07:15-08:15
MEET the PROFESSOR
LALEZAR HALL
Attilio Orazi, New York, NY, USA
Myeloproliferative Neoplasms: Practical Tips Based on Personal Experience.
08:30-17:00
08:30-08:45
08:45-09:45
08:45-09:15
EAHP LYMPHOMA SYMPOSIUM
MAIN HALL
WELCOME WORDS
Isinsu Kuzu, Ankara, Turkey
Elias Campo, Barcelona, Spain
EAHP LYMPHOMA SYMPOSIUM SESSION 1
07:15-08:15
19 OCTOBER 2014, SUNDAY
KEY NOTE LECTURE 1
Chairs: Isinsu Kuzu, Elias Campo
Immunology of normal B-cell differentiation
Michel Cogné, Limoges, France
09:15-09:45
TOPIC LECTURE 1
Chair: Leticia Quintanilla-Fend
Follicular Lymphoma: How many diseases?
Andreas Rosenwald, Würzburg, Germany
09:45-10:45
PROFFERED PAPERS SESSION 1
Chairs: Patty Janssen, Rafael Andrate
09:45-10:00
[OP-LYMP-06]
Atypical CD10-negative and/or BCL2-negative Follicular lymphoma are genetically
heterogeneous and comprise a subset with 1p36 deletion
Vanessa Szablewski1, Maryse Baia2, Christian Bastard3, Christine Terre4, Teresa Marafioti5,
Jean Michel Picquenot6, Claire Glaser7, Marie Hélène Delfau Larue8, Jehan Dupuis9, Corinne Haioun9,
Philippe Gaulard10, Christiane Copie Bergman10
1
Département de Biopathologie Cellulaire et Tissulaire des Tumeurs, CHU Montpellier, Hôpital Gui De
Chauliac, Montpellier, France
2
INSERM, Unité 955, Equipe 9, 94010 Créteil, France
3
INSERM U918, Centre Henri Becquerel, Rouen, France
4
Laboratoire de cytogénétique hémato-oncologique, Centre Hospitalier de Versailles, Le Chesnay, France
5
Department of Histopathology, University College Hospital London, UK
6
Département de Biopathologie Intégrée du Cancer, Centre Henri Becquerel, Rouen, France
7
Hôpital Mignot, service d’Anatomie Pathologique, Versailles, France
8
Service d’Immunologie Biologique, Hôpital Henri Mondor, Créteil, France
9
Unité Hémopathies Lymphoïdes, Hôpital Henri Mondor, Créteil, France
10
Département de Pathologie, Hôpital Henri Mondor, Créteil, France
| 17-22 October 2014
| EAHP - 2014
|
21
[OP-LYMP-17]
Recurrent mutations of NOTCH genes in follicular lymphoma identify a distinctive subset
of tumors
Kennosuke Karube1, Daniel Martínez1, Cristina Royo1, Magda Pinyol1, Paola Castillo1, Alexandra Valera1,
Anna Carrió1, Dolors Costa1, Dolors Colomer1, Maite Cazorla1, Daniel Esteban1, Andreas Rosenwald2,
German Ott3, Eva Giné1, Armando López Guillermo1, Elias Campo1
1
IDIBAPS, Hospital Clinic, Universitat de Barcelona, Barcdlona, Spain
2
Institute of Pathology, University of Würzburg, Würzburg, Germany
3
Department of Clinical Pathology, Robert-Bosch-Krankenhaus, and Dr. Margarete Fischer-Bosch Institute of
Clinical Pharmacology, Stuttgart, Germany
10:15-10:30
[OP-LYMP-19]
CD23-positive diffuse nodal follicular lymphoma: a distinct variant of follicular lymphoma
mimicking nodal marginal zone lymphoma
Keegan Barry Holson1, Charles Ma2, Lizalynn Dias2, Jane Houldsworth2, Russell K. Brynes1,
Imran N. Siddiqi1
1
Department of Pathology, University of Southern California Keck School of Medicine, Los Angeles,
California, USA
2
Cancer Genetics, Inc., Rutherford, New Jersey, USA
10:30-10:45
[OP-LYMP-20]
Novel Markers for the Diagnosis of Paediatric Follicular Lymphoma
Ayse U. Akarca1, Hasan Rizvi2, Claudio Agostinelli3, Alan Ramsay4, Maria Pane Foix4, Joan Somja4,
James Wilton1, Vishvesh H Shende1, Brunangelo Falini5, Stefano A Pileri3, David Linch6, Stephen Daw7,
Teresa Marafioti8
1
Department of Pathology, University College London, UK
2
Department of Cellular Pathology, Barts Health NHS Trust, London, UK
3
Section of Haematopathology, Department of Haematology and Oncological Sciences “Seràgnoli”, S.
Orsola-Malpighi Hospital, University of Bologna, Italy
4
Department of Cellular Pathology, University College Hospital London, UK
5
Institute of Hematology, University of Perugia, Perugia, Italy
6
Department of Haematology, University College London Cancer Institute, London, UK
7
Children and Young People’s Cancer Service, University College Hospital London, London
8
NIHR UCLH/UCL Biomedical Research Centre London, UK ‘;’ Department of Pathology, University College
London, UK
th
10:00-10:15
10:45-11:15
11:15-12:15
11:15-11:45
KEY NOTE LECTURE 2
Chair: Miguel Angel Piris
BCR activation as an oncogenic mechanism in lymphomas
Louis M. Staudt, Bethesda, MD, USA
11:45-12:15
TOPIC LECTURE 2
Chair: Steven H. Swerdlow
Plasmacytic differentiation in small B cell lymphomas other than extranodal MZL
Eric D Hsi, Cleveland, OH, USA
12:15-12:45
PROFFERED PAPERS SESSION 2A
Chairs: Maria Calaminicci, Frank Kuo
12:15-12:30
[OP-LYMP-14]
BCL6 protein expression and BCL6 chromosomal breaks in nodal marginal zone lymphoma
with diagnostic implications
Michiel Van Den Brand, Patricia Groenen, Konnie Hebeda, Han Van Krieken
Radboud university medical center, Nijmegen, the Netherlands
22
| EAHP - 2014
| 17-22 October 2014
Iris Miedema1, Nathalie Hijmering1, Jan Paul De Boer2, Olga Balagué Ponz3, Jacqueline Cloos4,
Sonja Zweegman4, Daphne De Jong1
1
VU University Medical Center, dept of Pathology, Amsterdam, the Netherlands
2
Netherlands Cancer Institute, Dept of Medical Oncology, Amsterdam, the Netherlands
3
Netherlands Cancer Institute, Dept of Pathology, Amsterdam, the Netherlands
4
VU University Medical Center, Dept of Hematology, Amsterdam, the Netherlands
12:45-13:00
13:00-14:30
Lunch Box
LUNCH & SATELLITE SYMPOSIUM
What the busy haematopathologist needs to know about Multicentric Castleman’s Disease
14:30-15:30
PROFFERED PAPERS SESSION 2B
Chair: Judith A Ferry, Jose Cabeçadas
[OP-LYMP-04]
Paediatric marginal zone lymphoproliferative disorder of the neck: a Haemophilus Influenzae
driven immune disorder?
Philip M. Kluin1, Ton Langerak2, Janetta Beverdam3, Lydia Visser1, Joop Van Baarlen4, King Lam5,
Kees Seldenrijk6,
Robby Kibbelaar7, Peter De Wit8, Ed Schuuring1, Stefano Rosati1, Arjan Diepstra1, Max M. Van Noessel9,
Jacco C. Hunting10,
Mels Hoogendoorn11, Ellen Van Der Gaag12, Eveline De Bont13, Hanneke C Kluin14, Jerome Lo Ten Foe15,
Adri Van Der Zanden3
1
Dept of Pathology and Medical Biology, University Medical Center Groningen, University of Groningen,
The Netherlands
2
Dept Immunology, Erasmus Medical Center Rotterdam, EMCR, Rotterdam, The Netherlands
3
Labmicta, section of Molecular Microbiology, Hengelo, The Netherlands
4
Dept Pathology, LPON, Hengelo, The Netherlands
5
Dept Pathology, Erasmus Medical Centre Rotterdam, EMCR, Rotterdam, The Netherlands
6
Dept Pathology, St Antonius Hospital, Nieuwegein, The Netherlands
7
Dept Pathology, Pathology Friesland, Leeuwarden, The Netherlands
8
Dept Pathology, Amphia Hospital, Breda, The Netherlands
9
Dept Oncology & Hematology, Sophia Children Hospital, Rotterdam, Netherlands
10
Dept Internal Medicine, St Antonius Ziekenhuis, Nieuwegein, The Netherlands
11
Dept Internal Medicine, Medisch Centrum Leeuwarden, The Netherlands
12
Dept Paediatrics, Zorggroep Twente, Hengelo, The Netherlands
13
Dept Paediatric Oncology & Hematology, University Medical Center Groningen, University of Groningen,
Netherlands
14
Dept Hematology, University Medical Center Groningen, University of Groningen, Netherlands
15
Dept Medical Microbiology, University Medical Center Groningen, University of Groningen, Netherlands
[OP-LYMP-08]
Extramedullary plasmacytomas of the upper aerodigestive tract are extranodal marginal
zone lymphomas with plasmacytic differentiation
12:30-12:45
[OP-LYMP-05]
KLF2 mutation is the most frequent somatic change in splenic marginal zone lymphoma and
identifies a subset with distinct genotype
Alexandra Clipson1, Ming Wang1, Laurence de Leval2, Margaret Ashton-Key3, Andrew Wotherspoon4,
George Vassiliou5,
Niccolo Bolli5, Sarah Moody1, Leire Escudero Ibarz1, Gunes Gundem6, Kim Brugger7, Anthony Bench8,
Mike Scott8,
Hongxiang Liu9, George Follows8, Eloy F. Robles10, Jose Angel Martinez Climent10, David Oscier11,
A James Watkins12, Ming-Qing Du13
1
Division of Molecular Histopathology, Department of Pathology, University of Cambridge, UK
2
Institute of Pathology, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland
3
Department of Cellular Pathology, Southampton University Hospitals National Health Service Trust,
Southampton, UK
4
Department of Histopathology, Royal Marsden Hospital, London, UK
| 17-22 October 2014
| EAHP - 2014
|
23
[OP-LYMP-24]
Detection of MYD88 L265P and CXCR4 mutations is a valuable tool for diagnosis of
lymphoplasmacytic lymphoma and identifies cases with high disease activity
Janine Schmidt, Natalie Schindler, Irina Bonzheim, Birgit Federmann, Falko Fend, Leticia Quintanilla Martinez
Institute of Pathology and Neuropathology, University Hospital Tübingen, Tübingen, Germany
[OP-LYMP-12]
NOTCH pathway disruption in a subset of HCV-related diffuse large B cell lymphoma: its
associations to prognosis and to a possible marginal zone derivation
Marco Lucioni1, Luca Arcaini2, Davide Rossi3, Marta Nicola1, Roberta Riboni1, Antonio Ramponi4,
Valeria Virginia Ferretti2, Maurizio Bonfichi2, Manuel Gotti2, Aldo Maffi1, Giorgio Alberto Croci1,
Mariarosa Arra1, Valeria Fiacacdori2, Marzia Varettoni2, Sara Rattotti2, Lucia Morello2, Elena Dallera1,
Gianluca Gaidano3, Mario Cazzola2, Marco Paulli1
1
Anatomic Pathology Unit, Department of Molecular Medicine, University of Pavia, Pavia, and Pathology
Unit, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy Scientifico (IRCCS) Policlinico San Matteo,
Pavia, Italy
2
Division of Hematology, Department of Molecular Medicine, University of Pavia and Fondazione Istituto di
Ricovero e Cura a Carattere Scientifico (IRCCS) Policlinico San Matteo, Pavia, Italy
3
Division of Hematology, Department of Translational Medicine, Amedeo Avogadro University of Eastern
Piedmont, Novara, Italy
4
Division of Pathology, Department of Health Science, Amedeo Avogadro University of Eastern Piedmont,
Novara, Italy
th
5
Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK; Department
of Haematology, Addenbrooke’s Hospital, Cambridge University Hospitals NHS Foundation Trust,
Cambridge, UK
6
Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK
7
Department of Molecular Genetics, Addenbrooke’s Hospital, Cambridge University Hospitals NHS Foundation
Trust, Cambridge, UK
8
Department of Haematology, Addenbrooke’s Hospital, Cambridge University Hospitals NHS Foundation
9
Molecular Malignancy Laboratory, Addenbrooke’s Hospital, Cambridge University Hospitals NHS Foundation
10
Division of Oncology, Center for Applied Medical Research CIMA, University of Navarra, Pamplona, Spain
11
Department of Haematology, Royal Bournemouth Hospital, Bournemouth, UK
12
Division of Molecular Histopathology, Department of Pathology, University of Cambridge, UK; Department
of Haematology, 13Addenbrooke’s Hospital, Cambridge University Hospitals NHS Foundation Trust,
Cambridge, UK
15:30-16:30
KARL LENNERT LECTURE
Lymphoma Classification: A Journey From Controversy to Consensus
Can learning from the past make us 10% happier?
Chair: Elias Campo
Nancy L. Harris, Boston, USA
16:30-17:00
Coffee break and Poster view
17:00-18:30
SATELLITE SYMPOSIUM
CD30 in haematopathology: More than a diagnosis
24
| EAHP - 2014
| 17-22 October 2014
MEET the PROFESSOR
MERCURY HALL
Attilio Orazi, New York, NY, USA
Myeloproliferative Neoplasms: Practical Tips Based on Personal Experience.
07:15-08:15
MEET the PROFESSOR
LALEZAR HALL
Marsha Kinney, San Antonio, TX, USA
Outliers: Uncommon Manifestations of Classical Lymphoid Entities
08:30-09:30
08:30-09:00
MAIN HALL
KEY NOTE LECTURE 3
Chair: Philip Kluin
Genetic and epigenetic alterations of small B-cell lymphoma
Jude Fitzgibbon, London, UK
09:00-09:30
TOPIC LECTURE 3
Chair: Eric Hsi
Mantle Cell Lymphoma, an aggressive disease?
Wolfram Klapper, Kiel, Germany
09:30-10:30
07:15-08:15
20 OCTOBER 2014, MONDAY
Chairs: Ayoma Atygalle, Kenosuke Karube
09:30-09:45
[OP-LYMP-10]
The G protein-coupled estrogen receptor 1 (GPER) contributes to the proliferation and
survival of mantle cell lymphoma
Martina Rudelius1, Elena Hartmann1, Hilka Rauert Wunderlich1, Wolfram Klapper2, German Ott3,
Andreas Rosenwald1
1
Institute of Pathology, Julius-Maximillians-University Wuerzburg, Wuerzburg, Germany
2
Institute of Pathology, University Hospital Schleswig-Holstein, Campus Kiel, Germany
3
Departement of Clinical Pathology, Robert-Bosch-Krankenhaus and Dr. M. Fischer-Bosch Institute of
09:45-10:00
[OP-LYMP-23]
IGHV mutational status and potential associations with SOX11 expression and survival in
mantle cell lymphoma
Megan O. Nakashima1, Xiaoxian Zhao1, Xianqian Li1, Lisa Durkin1, Brian T. Hill2, Kai Fu3, Raymond Lai4,
Eric D. Hsi1
1
Laboratory Medicine, Cleveland Clinic, Cleveland, USA
2
Hematology Oncology, Cleveland Clinic, Cleveland, USA
3
Pathology and Microbiology, University of Nebraska, Omaha, USA
4
Laboratory Medicine and Pathology, University of Alberta, Edmonton, Canada
10:00-10:15
[OP-LYMP-16]
Genome-wide Analysis of Enhancer Acetylation in DLBCL and Mantle Cell Lymphoma
Russell J. H. Ryan1, Cem Sievers1, Jasleen Kaur1, Holly Whitton2, Robbyn Issner2, Charles Epstein2,
Bradley E. Bernstein1
1
Department of Pathology, Massachusetts General Hospital, Boston, Massachusetts, United States
2
Epigenomics Initiative, Broad Institute of Harvard University and M.I.T, Cambridge, Massachusetts,
United States
| 17-22 October 2014
| EAHP - 2014
|
25
[OP-LYMP-22]
Recurrent gene mutations in chronic lymphocytic leukemia: results from the REM clinical trial
Julia Gonzalez Rincon1, Sagrario Gomez1, Carol Lozano1, Miguel Piris Villaespesa1, Marcos Gonzalez2,
Eduardo Anguita3, Rosa Collado4, Felix Carbonell4, Raul Cordoba5, Antonia Rodriguez6, Nerea Martinez7,
Miguel Angel Piris8, José Antonio Garcia Marco9, Margarita Sanchez Beato1
1
Instituto Investigación Sanitaria Puerta de Hierro, Madrid, Spain
2
Hospital Universitario Salamanca-Centro Investigación Cancer, Salamanca, Spain
3
Hospital Clinico de Madrid, Spain
4
Consorcio Hospital General Universitario de Valencia, Spain
5
Hospital Universitario Infanta Sofia, Madrid, Spain
6
Hospital Universitario 12 de Octubre, Madrid, Spain
7
Instituto de Investigación Marqués de Valdecilla, Santander, Spain
8
Hospital Universitario Marques de Valdecilla- Instituto de Investigación Marqués de Valdecilla, Santander, Spain
10:30-11:00
11:00-11:30
11:00-11:30
KEY NOTE LECTURE 4
Chair: Philippe Gaulard, Muhit Ozcan
Treatment of B-cell lymphoma in the era of personalized medicine
Gilles Salles, Lyon, France
11:30-12:30
Chair: Philippe Gaulard, Nukhet Tuzuner
th
10:15-10:30
11:30-11:45
[OP-LYMP-02]
Biological and therapeutic relevance of the BRAF-MEK-ERK pathway in Hairy Cell Leukemia
Valentina Pettirossi°1, Alessia Santi°1, Elisa Imperi1, Guido Russo1, Alessandra Pucciarini1, Barbara Bigerna1,
Elisabetta Fortini1, Roberta Mannucci2, Gianluca Schiavoni1, Ildo Nicoletti2, Maria Paola Martelli1,
Ludger Klein Hitpass3, Brunangelo Falini^1, Enrico Tiacci^1
1
2
Institute of Internal Medicine and Oncologic Sciences, University of Perugia, Perugia, Italy
3
Biochip Laboratory, Institute for Cell Biology - Tumor Research, University of Duisburg-Essen Medical
School, Essen, Germany
11:45-12:00
[OP-LYMP-21]
The role of AKT/mTOR cascade in hairy cell leukaemia: interaction with BRAF signalling
pathway and prognostic significance
Georgia Levidou1, Eleftheria Lakiotaki1, Maria K. Angelopoulou2, Gerasimos A. Pangalis3,
Theodoros Vassilakopoulos2, Angelica A. Saetta1, Athanasia Sepsa1, Ilenia Chatziandreou1,
Gabriella Gainaru2, Pagona Flevari2, Sotirios Sachanas3, Maria Moschogiannis3, Christina Kalpadakis4,
Vasilis Milionis1, Panayiotis Panayiotidis5, Maria Dimopoulou2, Eleni Plata2, Konstantinos Konstantopoulos2,
Efstratios Patsouris1, Penelope Korkolopoulou1
1
Department of Pathology, University of Athens, Medical School, Greece
2
Department of Hematology and Bone Marrow Transplantation, University of Athens, Medical School, Greece
3
Department of Hematology, Athens Medical Center, Psychikon Branch, Greece
4
Department of Hematology, University of Crete, Heraklion, Greece
5
1st Department of Propedeutic Internal Medicine, University of Athens, Medical School, Greece
12:00-12:15
[OP-LYMP-07]
Blastic plasmacytoid dendritic cell neoplasm: molecular high-through-put technologies
shed new light on its histogenesis, pathobiology and therapeutic options
Stefano Aldo Pileri1, Valentina Tabanelli1, Federica Melle1, Antonella Laginestra1, Claudio Agostinelli1,
Emilio Berti2, Lorenzo Cerroni3, Fabio Facchetti4, Fabio Fuligni1, Raul Rabadan5, Pier Paolo Piccaluga1,
Maria Rosaria Sapienza1
1
Unit of Haematopathology, Bologna University School of Medicine, Bologna, Italy
2
Dermatology Unit, University of Milan-La Bicocca, Milan, Italy
3
Department of Dermatology, University of Graz, Graz, Austria
4
Pathology Section, Department of Molecular and Translational Medicine, University of Brescia, Brescia, Italy
5
Department of Biomedical Informatics and Center for Computational Biology and Bioinformatics, Columbia
University, New York, NY 10032, USA
26
| EAHP - 2014
| 17-22 October 2014
Annette M Staiger1, Heike Horn1, Matthias Vöhringer2, Ulrich Hay3, Elias Campo4, Andreas Rosenwald5,
German Ott1, M. Michaela Ott6
1
Department of Clinical Pathology, Robert-Bosch-Krankenhaus, and Dr. Margarete Fischer-Bosch-Institute of
2
Department of Internal Medicine II, Hematology and Oncology, Robert-Bosch-Krankenhaus, Stuttgart,
Germany
3
Department of Ear, Nose and Throat Surgery, Marienhospital, Stuttgart, Germany
4
Hematopathology Unit, Pathology Department, Hospital Clínic and University of Barcelona, Institute of
Biomedical Research August Pi i Sunyer (IDIBAPS), Barcelona, Spain
5
Institute of Pathology, University of Würzburg, Würzburg, Germany and Comprehensive Cancer Center
Mainfranken (CCCM)
6
Institute of Pathology, Caritas-Hospital, Bad Mergentheim, Germany
12:30-12:45
12:45-14:15
Lunch Box
Common B-cell lymphomas: CLL/SLL and DLBCL
14:15-15:30
[OP-LYMP-18]
Diffuse Large B-Cell Lymphomas of Immunoblastic Type are a Major Reservoir for MYC-IgH
Translocations
12:15-12:30
POSTER DISCUSSION
Elena Sabattini, Bologna, Italy
Maurilio Ponzoni, Milan, Italy
John Goodlad, Edinburgh, UK
15:30-16:00
Coffee Break
16:00-16:15
DAVID Y MASON AWARD
Chairs: Elias Campo, Teresa Marafioti
16:15-17:15
DAVID Y MASON LECTURE
Chair: Elaine S. Jaffe
Mutational Processes in Human Cancer
Michael R. Stratton, Hixton, UK
17:15-18:15
GENERAL ASSEMBLY
| 17-22 October 2014
| EAHP - 2014
|
27
07:15-08:15
MEET the PROFESSOR
LALEZAR HALL
Outliers: Uncommon Manifestations of Classical Lymphoid Entities
07:15-08:15
MEET the PROFESSOR
MERCURY HALL
“May be Hodgkin Lymphoma, maybe not”
08:30-09:30
08:30-09:00
MAIN HALL
TOPIC LECTURE 5
Chair: Stefano Pileri
Molecular genetics of in the transformation of indolent lymphoma
Randy Gascoyne, Vancouver, Canada
th
21 OCTOBER 2014, TUESDAY
09:00-10:30
Chairs:Carmen Lome-Maldonado, Kikkeri Naresh
09:00-09:15
[OP-LYMP-03]
Diffuse large B-cell lymphoma with high expression of MYC and BCL2 shows evidence of
active B-cell receptor signaling by quantitative immunofluorescence
Agata M. Bogusz1, Alexandra E. Kovach2, Long P. Le2, Richard H. G. Baxter3, Aliyah R. Sohani2
University of Pennsylvania, Department of Pathology and Laboratory Medicine, Philadelphia, PA, USA
2
Massachusetts General Hospital, Department of Pathology, Boston, MA, USA
3
Yale University, Department of Chemistry, New Haven, CT, USA
1
09:15-09:30
[OP-LYMP-01]
Identification of single-nucleotide variants by RNA sequencing in endemic Burkitt Lymphoma
Pier Paolo Piccaluga1, Francesco Abate2, Maria Antonella Laginestra1, Giulia De Falco3, Maryam Etebari1,
Fabio Fuligni1, Maura Rossi1, Sakellarios Zairis2, Cristiana Bellan3, Lorenzo Leoncini3, Raul Rabadan2,
Stefano Aldo Pileri1
1
Department of Experimental, Diagnostic, and Specialty Medicine, University of Bologna, Bologna, Italy
2
Columbia University College of Physicians and Surgeons, New York, NY-USA
3
Department of Medical Biotechnologies, University of Siena, Siena, Italy
09:30-09:45
[OP-LYMP-15]
The pressure of the antigens in the pathogenesis of BL. Evidence from NGS analysis of
BCR
Cristiana Bellan1, Teresa Amato1, Pier Paolo Piccaluga2, Francesco Abate2, Giulia De Falco1,
Maria Raffaella Ambrosio1, Raul Rabadan3, Stefano Pileri2, Lorenzo Leoncini1
1
Anatomical Pathology Section, Department of Medical Biotechnology, University of Siena, vie delle Scotte,
6, 53100, Siena, Italy
2
Molecular Pathology Laboratory, Department of Experimental, Diagnostic, and Specialty Medicine, Bologna
University Medical School, Unit of Hematopathology, S. Orsola Malpighi Hospital, Via Massarenti 9, 40138
Bologna, Italy.
3
Department of Biomedical Informatics, College of Physicians and Surgeons, Columbia University, New
York, New York, USA
28
| EAHP - 2014
| 17-22 October 2014
Benjamin Rengstl1, Frederike Schmid1, Christian Weiser1, Claudia Döring1, Tim Heinrich1, Kathrin Warner2,
Petra S. A. Becker3, Christian Seidl3, Hinrich Abken2, Ralf Küppers4, Martin Leo Hansmann1, Sebastian Newrzela1
1
Dr. Senckenberg Institute of Pathology, Medical School, Goethe-University of Frankfurt, 60590 Frankfurt am
Main, Germany
2
Department I of Internal Medicine, Medical School, University of Cologne, 50937 Cologne, Germany
3
Institute for Transfusion Medicine and Immunohematology, Red Cross Blood Donor Service, BadenWürttemberg-Hessen, 60528 Frankfurt am Main, Germany
4
Institute of Cell Biology (Cancer Research), Medical School, University of Duisburg-Essen, 45122 Essen, Germany
10:00-10:15
[OP-LYMP-11]
Follicular cytotoxic CD8 T-cells (CD8TFC) sharing functional similarities with CD4TFH
cells are present in classical hodgkin’s lymphoma tissues displaying a specific “mixed
nodularity” pattern
Luc Xerri, Suong Le, Sylvaine Just Landi, Françoise Gondois Rey, Samuel Granjeaud, Daniel Olive
Centre de Recherche en Cancérologie de Marseille, INSERM U1068 and Institut Paoli-Calmettes, Marseille,
France Marseille, France
10:15-10:30
[OP-LYMP-09]
Somatic mutation screening using archival formalin-fixed paraffin-embedded tissues by
Fluidigm Access Array multiplex PCR and Illumina MiSeq sequencing
Ming Wang1, Leire Escudero Ibarz1, Sarah Moody1, Naiyan Zeng1, Alexandra Clipson1, Yuanxue Huang1,
Xuemin Xue1, Nicholas F Grigoropoulos1, Sharon Barrans2, Lisa Worrillow2, Tim Forshew3, Francesco Marass3,
Nitzan Rosenfeld3, Jing Su3, Andrew Firth1, Howard Martin4, Andrew Jack2, Kim Brugger4, Ming Qing Du1
1
Department of Pathology, University of Cambridge, Cambridge, UK
2
Haematological Malignancy Diagnostic Service, St. James’s Institute of Oncology, Leeds, UK
3
Cancer Research UK Cambridge Institute, Li Ka Shing Centre, University of Cambridge, Robinson Way,
Cambridge, UK
4
Department of Molecular Genetics, Addenbrooke’s Hospital, Cambridge University Hospitals NHS
Foundation Trust, Cambridge, UK
10:30-11:00
[OP-LYMP-13]
Unraveling the role of CD4 T cells in Hodgkin lymphoma
09:45-10:00
Coffee Break
LYMPHOMA WORKSHOP
11:00-13:00
EAHP LYMPHOMA WORKSHOP SESSION 1
Plasmacytic differentiation in small B-cell lymphomas
Chairs: Steven H. Swerdlow, Elias Campo
Introduction
LYWS 80
Lakshmi Venkatraman, Belfast, Northern Ireland, UK
LYWS 267
Bachir Alobeid, New York, NY, USA
LYWS 190
Elaine S Jaffe, Bethesda, USA
LYWS 58
Isinsu Kuzu, Ankara, Turkey
LYWS 150
LYWS 89
Inmaculada Ribera-Cortada, Barcelona, Spain
LYWS 46
Margaret Rose Ashton-Key, Southampton, UK
LYWS 40
LYWS 49
Filiz Sen, NewYork, NY, USA
PANEL REVIEW
LYWS0045, LYWS0057, LYWS0078, LYWS0082, LYWS0085, LYWS0104, LYWS0107,
LYWS0311, LYWS0312, LYWS0315, LYWS0316, LYWS0326
13:00-14:00
Lunch
| 17-22 October 2014
| EAHP - 2014
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29
th
14:00-16:00
Marginal zone lymphomas: Splenic, nodal and MALT type
Chairs: Ahmet Dogan, Isinsu Kuzu
Introduction
SPLENIC
LYWS 28
LYWS 81
LYWS 178
LYWS 72
Philipp W. Raess, Pennsylvania USA
Fina Climent, Barcelona, Spain
Miguel A Piris, Santander, Spain
Kennosuke Karube, Barcelona, Spain
NMZ
LYWS 302
LYWS 288
Serap Isiksoy , Eskişehir, Turkiye
Monika Klimkowska, Stockholm, Sweeden
MALT
LYWS199
Claudiu V. Cotta, Cleveland, USA
LYWS 175
April Chiu, New York, USA
16:00-16:30
Coffee Break
16:30-18:00
Mantle Cell Lymphoma
Chairs: Birgitta Sander, Leticia Quintanilla-Fend
LYWS 108
Larissa Sena T. Mendes, London, UK
LYWS 61
Amy Evelyn Rich, Orlando, USA
LYWS 33
Megan O. Nakashima, Cleveland, USA
LYWS 115
Sarah E. Gibson, Pittsburgh, USA
LYWS 282
William R. Macon, Rochester, USA
LYWS 134
Randy D. Gascoyne, Vancouver, Canada
PANEL REVIEW
LYWS0328
30
| EAHP - 2014
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MAIN HALL
Paediatric lymphomas
Chairs: Leticia Quintanilla-Fend, Birgitta Sander
Introduction
LYWS 189
Gerald Wertheim, Philadelphia, USA
LYWS 34
Metin Taskin, New Jersey, USA
LYWS 166
Arianna Di Napopli, Rome, Italy
LYWS 250
Elaine S Jaffe, Bethesda, USA
LYWS 272
Thierry Molina, Paris, France
LYWS 324
Abner Louissaint Jr., Boston, USA
LYWS 228
Maria A. Pletneva, Michigan, USA
PANEL REVIEW
LYWS0042, LYWS0084, LYWS0100, LYWS0161, LYWS0177, LYWS0180, LYWS0183, LYWS0220,
LYWS0229, LYWS0234, LYWS0243, LYWS0245, LYWS0254, LYWS0276, LYWS0294, LYWS0323
10:00-10:30
10:30-13:00
Coffee Break
Follicular lymphoma and CLL/SLL
Chairs: German Ott, Luc Xerri
Introduction
LYWS67
Carmen Lome-Maldonado, Mexico City, Mexico
LYWS 127
Luc Xerri, Marseille, France
LYWS 216
Xiaohui Zhang, Tampa, USA
LYWS 192
Philip Kluin, Groningen, Netherlands
LYWS 24
Rebecca King, Rochester, USA
LYWS 237
Phillipe Gaulard, Créteil, France
LYWS 193
Jagmohan Singh Sidhu, Johnson City, USA
LYWS 36
Ahmet Dogan, NY, USA
Summary and Conclusions FL
LYWS 232
Madhu Menon, Detroit, USA
LYWS 263
Mufaddal TaherMoonim, London, UK
Summary and Conclusions CLL
PANEL REVIEW
LYWS0280, LYWS0287
13:00-14:00
14:00-16:00
08:00-10:00
22 OCTOBER 2014, WEDNESDAY
Lunch
Progression in small B-cell lymphomas and composite lymphomas
Chairs: John Chan, Stephan Dirnhofer
Introduction
LYWS 188
Karthik Ganapathi, Bethesda, USA
LYWS 196
Prabjot Kaur, Florida, USA
LYWS 71
Stephan Dirnhofer, Basel, Switzerland
LYWS 113
Leticia Quintanilla-Fend, Tübingen, Germany
LYWS 98
Igor Schliemann, Huddinge, Sweden
LYWS 133
Daisy Alapat, Little Rock, USA
LYWS 173
Michael G. Bayerl, Pittsburgh, USA
PANEL REVIEW
LYWS0265, LYWS0274, LYWS0275, LYWS0278, LYWS0301, LYWS0317
16:00-16:30
FINAL WORDS ABOUT THE WORKSHOP and CLOSING REMARKS
Elias Campo
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31
DATE : OCTOBER 19, 2014 SUNDAY
TIME
: 13:00-14:30
PLACE : Main Hall
What the busy haematopathologist needs to know about Multicentric Castleman’s Disease
Chairman
: Ahmet Dogan (USA)
Scientific chair
: Ahmet Dogan (USA)
Faculty
: Falko Fend (Germany), Pier-Luigi Zinzani (Italy)
PROGRAM
Introduction to Multicentric Castleman’s Disease
Ahmet Dogan
How to overcome challenges of diagnosis
Falko Fend
th
S AT EL L I T E S YM P OS I UM S
SATELLITE SYMPOSIUMS
Interactive case study discussions
Ahmet Dogan and Falko Fend
Treatment options: moving ahead in Multicentric Castleman’s Disease
Pier-Luigi Zinzani
Summary and close
Ahmet Dogan
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SATELLITE SYMPOSIUM
Support for this educational activity is provided by Millennium: The Takeda Oncology Company and Takeda.
Title of symposium: CD30 in haematopathology: more than a diagnosis
Chair
: Stefano Pileri, University of Bologna, Bologna, Italy
Speakers : Mine Hekimgil, Ege University, Izmir, Turkey
Timothy Illidge, University of Manchester, Manchester, UK
Aysegul Uner, Hacettepe University, Ankara, Turkey
Learning objectives of the EAHP satellite symposium
After having participated in the satellite symposium, the pathologist will:
1) Understand the importance of CD30 detection in diagnosis and treatment
2) Apply the best practice to detect CD30 in malignant tissues, recognize the pitfalls in CD30
detection and potential solutions
3) Evaluate recent data from clinical trials using CD30-targeting therapeutics on relapsed/refractory
HL and sALCL, which focuses on brentuximab vedotin, and outline the clinical relevance for other
haematological malignancies
PROGRAM
17:00-17:05 Welcome and introduction (Prof. Stefano Pileri)
17:05-17:15 CD30 staining: Getting the results right (Prof. Stefano Pileri)
17:15-17:30 The need for CD30 detection: A haematologist’s perspective (Prof. Timothy Illidge)
17:30-17:50 The challenges of CD30 detection: Interactive case study 1 (Prof. Mine Hekimgil)
17:50-18:10 The challenges of CD30 detection: Interactive case study 2 (Prof. Aysegul Uner)
18:10-18:25 Panel discussion and Q&A
18:25-18:30 Closing (Prof. Stefano Pileri)
| 17-22 October 2014
| EAHP - 2014
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DATE : OCTOBER 19, 2014 SUNDAY
TIME
:17:00-18:30
PLACE : Main Hall
33
PROGRAM
Common B-cell lymphomas: CLL/SLL and DLBCL:
Satellite Symposium of 17th Meeting of the European Association of Haematopathology
This session focuses on the utility of novel approaches for aiding in the diagnosis and stratification of
mature B-cell leukemias and lymphomas. It includes two Speakers Yaso Natkunam MD, PhD, Samantha
Kendrick, PhD, and Leigh Ann Henricksen, PhD., and a moderated discussion led by Lisa Rimza, MD and
Teresa Marafioti, MD. Dr. Natkunam will present data focused on reactive vs. neoplastic B-cell proliferations.
Dr. Kendrick will present data on differences in clones in bcl-2 antibodies, and Dr. Henrickson will present
data on the utility of chromagenic in situ hybridization (CISH) CLL probes.
Introduction
Peter Banks, M.D.
Past President SH
Ventana Medical Systems, Inc.
Tucson, Arizona, USA
th
DATE : OCTOBER 20, 2014 MONDAY
TIME
:12:45-14:15
PLACE : Main Hall
Immunoblasts or Hodgkin Cells? That is the Question
Yaso Natkunam, MD, PhD
Professor of Pathology
Stanford University Medical Center
Stanford, California, USA
Chromogenic Methods for Detecting Chromosomal Alterations in CLL Clinical Specimen
Leigh A. Henricksen, Ph.D.
Ventana Medical Systems, Inc.
BCL2 in diffuse large B-Cell Lymphoma: Old marker, New Applications
Samantha Kendrick, PhD
The University of Arizona
Moderated discussion
Lisa Rimsza, M.D.
Professor of Pathology
The University of Arizona Medical Center
Teresa Marafioti, MD
University College London Hospitals
London, UK
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AYS E U. AKAR CA
Ayse earned her BSc (Hons) in Biology from University of Çukurova in Turkey in 2004.
She began her career in haematology at University College London under supervision
of Prof.Marafioti.
Ayse’s research is mainly on characterisation and identification of new molecules
as diagnostic and prognostic markers for haematological disorders. Her research is
published in British Journal of haematology and American Journal surgical Pathology.
Ayse’s multiple colour immunostaining of formalin fixed paraffin embedded human
tonsil tissue which showed the protein expression of up to 5 colours was selected and
exhibited for the PHOTO:synthesis photography competition.
JO N A STER
My lab studies Notch signaling in cancer, work that dovetails with my role at Brigham
and Women’s Hospital, where I lead the Hematopathology division. Notch receptors act
in a signaling pathway that controls fundamental cellular processes in a context-specific
fashion. My lab has led or collaborated on work that has produced the first mouse model
of Notch leukemia; demonstrated that Notch signals induce T cell development from
bone marrow progenitors; demonstrated that T-ALL cells depend on Notch signaling for
growth; identified frequent Notch1 mutations in T-ALL; solved key Notch structures at
high resolution; identified Myc and mTOR as targets of leukemogenic Notch signaling;
developed the first selective Notch receptor inhibitors; viii) reported genome-wide
Notch1 binding patterns in cancer cell genomes; and described Notch1 loss-of-function
mutations in squamous cell carcinomas. I lead multi-institutional NCI P01 and LLS SCOR grants that are focused
on translating Notch-directed therapeutics into clinical practice.
BIOSKETCH & PHOTO OF INVITED SPEAKERS & ORAL PRESENTERS
B IO S K E T C H & P HO TO OF INVIT ED SPEAK ER S &
O R A L P RE SENTE RS
A D A M B AGG
Adam Bagg is a Professor of Pathology and Laboratory Medicine at the University of
Pennsylvania, where he serves as Director of Hematology, Medical Director of Clinical
Cancer Cytogenetics and Interim Director of Hematopathology. For 10 years he was
Director of the Leukemia and Lymphoma Society of America-funded Minimal Residual
Disease Core Facility of the Center for Immunotherapy at Penn. He has been the
recipient of numerous research grants, including some from the NIH. In recognition of
his teaching endeavors he was awarded the Kevin E. Salhany MD Award for Excellence
in Clinical Teaching in 2000 and again in 2010. For the past 5 years (2010-2014), he
was voted by his peers as one of Philadelphia’s “Top Doctors”. US News and World
Report has noted him to be in the top 1% of doctors in the United States. In 2011, he
was elected to Council/Board of Directors of USCAP. He has lectured extensively nationally (including at USCAP,
ASCP, AMP, CAP, ACLPS, AACC, ISLH and ASH) and internationally. He has over 140 publications, including
peer-reviewed articles, invited reviews and textbook chapters, most in the realm of the molecular pathology of
hematologic malignancies. A publication that he was a coauthor on (CART19 T-cells to treat CLL) was the most
downloaded article in Science Translational Medicine for 2011. He is an Associate Editor of the Journal of Molecular
Diagnostics and on the Editorial Board of Advances in Anatomic Pathology.
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35
CRI S TI ANA B ELLAN
Cristiana Bellan, MD, PhD, is Researcher of Molecular Pathology, at the Department of
Medical Biotechnologies, Pathological Anatomy Section, University of Siena, Italy. She has
experience in immunoglobulin and TCR genes analysis with a particular focus on Burkitt
lymphoma. She has didactic activities at University of Siena: Lecturer in Pathological
Anatomy for degree course in Medicine and Surgery and in Biotechnology for Human
Health.
She performs her diagnostic activity at the “DAI servizi” of “ Azienda Ospedaliera
Universitaria Senese”. Cristiana Bellan has also conducted research and diagnostic
activities at the following Institutes and departments:
 Molecular Pathology Lab of Pathology, Anatomy and Cell Biology Institute, Thomas Jefferson University,
Philadelphia, PA (USA), (from February 1999 to august 2000);
 Molecular Pathology Lab, Institut fur Pathologie, Universitatasklinikum Benjamin Franklin, Freie Universitat
Berlin (from January 2003 to February 2003);
 Alma Mater Studiorum, Università di Bologna, Dpt. Di Ematologia e scienze Oncologiche “Le A. Seragnoli”,
Sezione di Anatomia, Istologia e Citologia Patologica “Marcello Malpighi”, Ospedale Bellaria, Bologna, Italy. (from
January 2011 to August 2011).
A GATA M . B OGU SZ
Agata Bogusz is an Assistant Professor in the Department of Pathology and Laboratory
Medicine at the University of Pennsylvania. Dr. Bogusz received herM.D. degree in 1999
and her and Dr. med. (DMSc) degree summa cum laude in 2003 from the RWTH Aachen
University in Germany. She earned a PhD degree in Pathology at the University of Chicago
in 2006 and was awarded the pre-doctoral grant from the Department of Defense and
Susan G. Komen foundation. She completed residency training in Anatomic Pathology at
UT Southwestern Medical Center at Dallas in 2009. After this she was a postdoctoral fellow
for 2 years at the Brigham and Women’s Hospital in the laboratory of Dr. Jeffery Kutok
and her work was focused on elucidating of a signature of active B cell receptor signaling
in diffuse large B cell lymphoma using quantitative immunofluorescence. Following her
postdoc Dr. Bogusz completed fellowship training in Hematopathology at Beth Israel and Deaconess Medical Center
in Boston, MA in 2012 and fellowship training in Molecular Genetic Pathology at Yale University in 2013. She is
board certified in Anatomic Pathology Hematopathology and Molecular Genetic Pathology by the American Board of
Pathology and in Pathology by the European Board of Pathology.
MI C HI EL VAN DEN B RAND
Michiel van den Brand is a pathology resident and PhD student at the Radboud university
medical center in Nijmegen, the Netherlands. His research aims to improve lymphoma
classification and our understanding of the pathogenesis of lymphomas. His current
work focuses on nodal marginal zone lymphoma, a lymphoma with a largely unknown
pathogenesis that is often difficult to diagnose. These studies are oriented towards the
practice of pathology and aim to provide novel tools for diagnosis, treatment prediction,
and prognosis.
A L EXA NDR A CLIPSON
Dr Alexandra Clipson is a research associate in the laboratory of Professor Ming-Qing Du in
the Department of Pathology at the University of Cambridge. Over the last three years her work
has focused on characterising the genetics of splenic marginal zone lymphoma (SMZL) using
next generation sequencing techniques. Alex obtained her PhD in Chemical Biology under the
guidance of Professor Michael Greaney at the University of Edinburgh, with a thesis titled “A
target-guided synthesis approach to the discovery of novel bivalent inhibitors of Glutathione
Transferases”. As part of her postgraduate studies, she gained an MSc in Life Sciences.
Alex graduated with an MChem in Chemistry from the University of York in 2006. During her
undergraduate studies, she spent a year working as a Medicinal Chemist with Merck.
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| EAHP - 2014
| 17-22 October 2014
Pr. Michel Cogné, MD, PhD, studied immunoglobulin (Ig) gene alterations in human disorders
and transcriptional regulation of Ig genes, successively in the Universities of Paris 6, Poitiers,
Harvard and Limoges. He contributed unravelling the gene alterations in heavy chain diseases
and connected them with aberrations in V(D)J recombination, somatic hypermutation and
class switching. He first showed that 3’ IgH enhancers control class switching and revealed
the conserved palindromic structure of the 3’ IgH regulatory region (Cogné et al, 1994,
Chauveau et al, 1996, Pinaud et al, 1998 and 2001, Wuerffel, 2007). His team showed the
role of 3’ IgH enhancers in lymphomagenesis (Truffinet 2007, Gostissa, 2010), somatic
hypermutation (Rouaud, 2013) and in a recently reported B cell death pathway through IgH
locus suicide recombination (Péron 2012). He also set up models of light chain deposition
diseases (Khamlichi 1995, Decourt 1999, Rengers 2000, Sirac 2006), and contributed original models of allelic exclusion,
interallelic switching, class-switched BCR function (Delpy 2004, Reynaud 2006, Sirac 2006, Duchez 2010, Laffleur, 2014).
FRÉ D É R IC DAVI
Professor of Hematology
Hôpital Pitié-Salpêtrière
Université Pierre et Marie Curie
Paris, France
Dr Frédéric Davi received his medical degree from Paris Université René Descartes
in 1987, and performed his residency training in Biological Hematology in Poitiers. He
then completed his PhD training in Immunology (working on immunoglobulin genes) at
the Saint-Louis Hospital in Paris. Thereafter he joined the laboratory of Jeff Sklar in the
Department of Pathology at the Brigham & Women’s Hospital in Boston (USA) for a postdoctoral fellowship. Upon his return in Paris at the Pité-Salpêtrière hospital in 1993, he created and developed the
Molecular Haematology Laboratory. He became head of the Department of Biological Hematology in 2012.
MI CHEL COGNÉ
Dr Frédéric Davi’s main research interest is the study of the repertoire of immunoglobulin genes in lymphoid neoplasia. He
has been one of the founders of the European Biomed-2 group, who standardized the analysis of antigen receptor genes for
clonality assesment in lymphoproliferations ; he is now member of the Board of the EuroClonality consortium. He is also one
of the founders of the IgCLL group, dedicated to the analysis of immunoglobulin repertoire in chronic lymphocytic leukemia.
BRUNA NGELO FALINI
Dr. Falini is the head of Institute of Hematology, University of Perugia, Italy. His basic
research has focused on the immunological and molecular characterization of lymphomas
and leukemia. He pioneered seminal contributions in the field, including the generation
of novel monoclonal antibodies for detection of lymphoma/leukemia associated genetic
lesions in paraffin sections, the construction and use of the first anti-CD30 immunotoxin
for the treatment of refractory/resistant Hodgkin’s disease, the characterization of ALK+
anaplastic large cell lymphoma, and the breakthrough discoveries of NPM1 mutations
in AML and BRAF-V600E in HCL. His work had a major clinical impact in the diagnosis
and prognostic stratification of AML and also in the development of a molecular targeted
therapy with BRAF inhibitors of refractory/relapsed HCL. Dr. Falini is the recipient of the
José Carreras award from the European Society of Hematology (2011) and the Karl Lennert Lecture Award from the
European Association for Hematopathology (2012).
JUD E FITZGIB B ON
Jude Fitzgibbon is Professor of Personalised Cancer Medicine in the Centre for HaematoOncology at the Barts Cancer Institute (http://www.bci.qmul.ac.uk/staff/item/fitzgibbon)
part of Queen Mary University of London. His main research interest is an incurable
B cell malignancy called Follicular Lymphoma, and gaining an understanding of
the genetic architecture and clonal evolution of this indolent disease, its switch to more
aggressive forms of lymphomas and defining actionable mutations that offer opportunities
to stratify treatment and optimisecare.
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37
RA ND Y D. GASCOYNE
Clinical Professor of Pathology, University of British Columbia
Hematopathologist, BC Cancer Agency
Medical Director – Provincial Lymphoma Pathology Program
Distinguished Scientist, BC Cancer Research Centre
Research Director, Centre for Lymphoid Cancer, BCCA
Department Head – Lymphoid Cancer Research, BCCRC
Randy D. Gascoyne is a Clinical Professor of Pathology at the University of British
Columbia, a Hematopathologist at the BC Cancer Agency (BCCA) and a Distinguished
Scientist at the BC Cancer Research Centre in Vancouver, Canada.
Dr. Gascoyne has more than 410 peer-reviewed publications, has co-authored > 430 abstracts at major meetings and
has written 20 book chapters. During his tenure at the BCCA he has been a principle investigator/co-investigator on
research grants totaling over $94,000,000. He served as Associate Editor of Haematologica from 2008 - 2012. He is
the pathology co-chair of the ECOG’s Lymphoma Committee. He is an active member of the International Lymphoma
Study Group (ILSG). He is currently the Research Director for the Centre for Lymphoid Cancers at the BC Cancer
Agency in Vancouver.
In 2011-12 Randy received several awards, including a Killam Research award in Science from the University of
British Columbia, establishing the first Clinical Faculty to be awarded such a prize. In late 2011 he was awarded an
Honorary Doctorate Degree (Docteur Honoris Causa) from the University of Paul Sabatier in Toulouse, France. In 2012
he received an Excellence in Research & Discovery from the Department of Laboratory Medicine at the University of
British Columbia. In 2014 he was listed by Thomson-Reuters ISI in the top 1% of influential scientific minds based on
citations and impact factors during the period 2002-2012 in the category of Clinical Medicine.
NA NCY LEE HARRIS
Dr. Harris is the Austin L. Vickery Professor of Pathology at Massachusetts General
Hospital and the Harvard Medical School and Editor of the Case Records of the
Massachusetts General Hospital for the New England Journal of Medicine. Having
trained in hematopathology in an age of multiple classifications of lymphomas, Dr. Harris
became interested in establishing consensus in the classification of lymphoid neoplasms.
Working first with the International Lymphoma Study Group and then with the World
Health Organization, Dr. Harris and colleagues published the Revised European American
Classification of Lymphoid Neoplasms (REAL) in 1994 and in 2001, published a WHO
Classification of Tumours of the Haematopoietic and Lymphoid Tissues. The classification
was updated with a 4th edition in 2008, and an interim update is planned for 2015. This
classification represents the first true international consensus on the classification of hematologic neoplasms, and
established a paradigm for arriving at consensus among pathologists and clinicians on disease classification.
ERI C H SI
Dr. Hsi is the Section Head of Hematopathology and Chair of the Department of
Laboratory Medicine at the Cleveland Clinic. He is medical director of the clinical flow
cytometry laboratory and program director for the Hematology Fellowship. His clinical
interests include diagnosis and classification of myeloid and lymphoid neoplasms. His
research laboratory focusses upon pre-clinical biomarker assessment related to targeted
therapeutics. Dr. Hsi serves as Pathology Committee chair and Vice Chair for Lymphoma
Biology for the ALLIANCE, and also sits on the Hematology Resource Committee for the
College of American Pathologists.
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| EAHP - 2014
| 17-22 October 2014
Leire Escudero Ibarz is currently a PhD student at Cambridge University’s Department of
Pathology. Her research in Prof. Ming Du’s lab is focused on the study of the genetic bases
of diffuse large B-cell lymphoma in order to assess their potential value in disease prognosis
and treatment stratification. Leire previously worked in Dr. Queelim Ch’ng’s lab in King’s
College London, studying the genetics of ageing (2012). She previously received a Master
of Science degree in Molecular Biology and Pathology of Viruses (Virology) from Imperial
College London (2011), where she joined Prof. Nicholas Mazarakis’ lab to investigate the
use of viral vectors for gene therapy. Prior to moving to the UK, Leire obtained a Bachelor
of Science degree in Biotechnology from Universitat Autònoma de Barcelona (2010), and
undertook a research exchange in Università degli Studi di Pavia (Italy) studying under
Prof. Pietro Speziale (2009).
D A PHNE DE JONG
After working for 19 years at the Netherlands Cancer Institute, Daphne de Jong recently
moved to the VU University Medical Center in Amsterdam, the Netherlands as Professor of
Haematopathology. Her field of interest and research activities focus on B-cell lymphomas
and on integrated molecular diagnostics for classification and treatment. For HOVON,
the Dutch national platform for clinical trials in hematology, she is involved in the broad
implementation of (molecular) biomarkers in targeted treatment.
KENNOSU K E K ARU B E
L E I RE ESCU DER O IB AR Z
Kennosuke Karube is a hematopathologists at the IDIBAPS, Hospital Clinic in Barcelona.
He graduated in medicine at the Kyushu University and received M.D. in Japan in 2000. In
2006, he received his Ph.D. under the co-supervision of Prof. Harada (Kyushu University,
Japan), Prof. Kikuchi (Fukuoka University, Japan) and Dr. Ohshima (Fukuoka University,
Japan). Upon finishing his doctorate, Dr. Karube obtained a contract as Research Fellow
of the Japan Society for the Promotion and continued his research in Prof. Ohshima’s
group at Kurume University and in Prof. Seto’s group at Aichi Cancer Center Research
Institute. He then obtained a contract as Research fellow of Uehara Foundation (Japan)
and went abroad in Spain to expand his research in the laboratory of Prof. Campo. His
main research area is to study the clinicopathological and molecular characteristics in
lymphoid malignancies.
REBE CCA KING
Rebecca King earned her MD from the University of Pennsylvania in Philadelphia, PA
in 2007. She then stayed at Penn for residency in Anatomic and Clinical Pathology.
Following residency, she completed a Hematopathology fellowship at the Mayo Clinic
in Rochester, MN. She then served as Assistant Professor in Pathology and Laboratory
Medicine at the University of Pennsylvania where she was primarily practicing pediatric
hematopathology at the Children’s Hospital of Philadelphia. In January 2014, she
accepted a faculty position at the Mayo Clinic, where she is currently an Assistant
Professor in the Division of Hematopathology.
| 17-22 October 2014
| EAHP - 2014
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39
MA RSHA C. KINNEY
Dr. Kinney received B.A (Molecular Biology, Vanderbilt University), M.S. (Biology, Abilene
Christian University), and M.D. (University of Texas, Southwestern Medical School)
degrees and completed residency, hematopathology, and pediatric immunology research
training at Vanderbilt University Medical Center. She is board certified in Anatomic and
Clinical Pathology and Hematology and is Professor and Director of the Division of
Hematopathology, University of Texas Health Science Center at San Antonio.
Her career has focused on diagnostic hematopathology (particularly T-cell and cutaneous
lymphomas) and teaching. She serves on the editorial boards of the American Journal of
Surgical Pathology and the American Journal of Clinical Pathology. She is the immediate
Past President of the Society for Hematopathology and serves on the EAHP Executive Committee. She received
the American Society for Clinical Pathology (ASCP) Mastership Designation and the ASCP Board of Certification’s
Distinguished Service Award. She serves on the Hematology Test Committee of the American Board of Pathology and
the American Society of Hematology Committee on Practice. She has authored numerous original research articles,
reviews, and book chapters and has made invited presentations at national and international conferences, courses,
and named lectures.
WOL FRAM KLAPPER
Wolfram Klapper is Professor and Head of Haematopathology, and Senior Physician in
the Department of Pathology, Haematopathology Section and Lymph Node Registry at
the University of Kiel, Kiel, Germany. After obtaining his MD from the University of Kiel in
2001, Professor Klapper remained at the University to complete a residency in Internal
Medicine. From 2002 to 2008 he was Assistant Doctor at the Department of Pathology,
University of Kiel. Professor Klapper is a member of the Steering Committees for the
Deutschen Low-Grade Lymphom-Studiengruppe (GLSG) and for the German paediatric
NHL study group (NHL-BFM). He is also Reference Pathologist for studies of lymphoma
within the Competence-Network ‘Maligne Lymphome’ (KML e.V.). Since 2007, Professor
Klapper has been Head of the Panel of Pathology within the ‘Mantelzell-LymphomNetzwerk’ (MCL network) and of the panel ‘Pediatric Lymphome-Pathology’ within the European Intergroup for
Childhood NHL (EICNHL). Current projects include the International Cancer Genom Project on germinal center
lymphomas and other molecular studies in the field of lymphoma.
GEORGIA LEVIDOU
Georgia Levidou graduated from the Medical School, University of Athens in 2005 (grade
“Excellent”). In 2008 she received a MSc degree in Biostatistics in the University of Athens/
University of Ioannina (grade “Excellent”), whereas in 2010 she appointed a PhD in the
University of Athens (grade “Excellent”). She obtained a special diploma in Anatomic and
Clinical Pathology in 2011. In 2012 she performed a three-month research fellowship in the
Department of Hematology and Oncology, University of Bologna. Currently, she is working
as a Research Associate (equivalent to Lecturer) in the First Department of Pathology,
University of Athens where she signs approximately 3,000 biopsies per year corresponding
to Hematopathological specimens, under the supervision of Prof.Korkolopoulou and Prof.
Patsouris, the latter being the head of the Department. Dr Levidou has published over 50
journal articles and her research interest includes lymphoma pathobiology as well as brain tumours and urothelial
bladder carcinoma molecular pathology.
40
| EAHP - 2014
| 17-22 October 2014
Armando López-Guillermo is currently a Senior Consultant in the Department of Hematology,
Hospital Clínic, Barcelona, Spain. He gained his degree in Medicine and Surgery from the
Medical School, University of Barcelona, Spain and worked as a Research Fellow at the
Lymphoma and Myeloma Department, MD Anderson Center, Houston, Texas, USA.
Dr López-Guillermo is a member of several working groups and scientific committees
including the Catalan Group for the Study of Lymphomas, the Executive Committee of the
International Follicular Lymphoma Prognostic Factors Project, the International Extranodal
Lymphoma Study Group, the European Mantle Cell Lymphoma Study Group and the
Leukemia and Lymphoma Molecular Profiling Project. He has authored more than 150
papers in the field of haematological malignancies.
MA RC O LU CIONI
Marco Lucioni, born September 4, 1973, Novara (Italy), is pathologist at the Anatomic
Pathology Unit, Fondazione IRCCS Policlinico San Matteo.
His scientific activity is focused on haematopathology, with special interests on
lymphomas and post-transplant lymphoproliferative disorders. His activity is documented
by several papers on international peer reviewed journals with impact factor, and by
several oral presentations at international and national meetings. He has cooperated in
the field of haematopathology with various institutions both in Italy and abroad. Member
of the European Association for Haematopathology (EAHP), member of the Italian
Lymphoma Study Group (GISL), member of the Italian Haematolymphopathology Group
(GIE); member of Italian Lymphoma Foundation (FIL) and Cutaneous Lymphoma Italian Study Group (GILC).
A RMA N DO LÓPEZ-GU ILLERM O
He is currently focused on the study of the oncogenetic potential of infectious agents in the genesis of
lymphoproliferative disorders and on the study of molecular genetics of nodal and exranodal lymphomas.
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JEA N- PHILIPPE M ER LIO
Professional Address
1) EA2406 and Faculty of Medicine. Université de Bordeaux, 146 rue Léo Saignat,
33076 BORDEAUX Cedex, France
Phone : (33) 5 57571028 – Fax :(33) 5 57571032
E-mail : jp.merlio@u-bordeaux2.fr
2) Service de Biologie des Tumeurs-Tumorothèque. CHU de Bordeaux. Hôpital HautLévêque, Avenue de Magellan, 33604 Pessac
Phone : (33) 5 57 65 66 39
E-mail : jp.merlio@u-bordeaux2.fr
Date of birth: May 16, 1957
CURRENT POSITION
Professor in Histology-Embryology-Cytogenetic (then Cytology-Histology) (PU/PH), since 1993
Institution University of Bordeaux
Head of Research Team EA 2406 Histology and Molecular Pathology of Tumours
Head of the Tumour Biology Laboratory of Bordeaux University Hospital
Institution CHU Bordeaux
Head of the Tumour Bank of Bordeaux University Hospital
PROFESSIONAL BACKGROUND
MEDICAL STUDIES
Medical studies
Residency in Pathology (1982-1986)
SPECIALTY TRAINING
Sabbatical 1 year Department of Molecular Biochemistry (Dir. H Persson), 1991-2.
Institution Karolinska Institutet, Stockholm
Assistant Lecturer in Pathology (AHU), 1986-1988
Assistant Lecturer in Histology, Embryology and Cytogenetic (AHU), 1988-90.
Lecturer in Cytology and Histology (MCU-PH), 1990-93.
UNIVERSITY DIPLOMAS
Medical Doctor (MD), 1983
CES d’Anatomie et Cytologie Pathologiques (Qualification in Pathology), 1984.
Master Degree in Genetic
1985
H.D.R. (French Board allowing to lead research projects) 1993
Physician Doctor (PhD) in Cell Biology and Genetic, 1996.
MEMBERSHIPS
• Member of the steering committee for clinical research of University Hospital of Bordeaux since 1996.
• Member of the INCa group of Tumor Genetic Regional Platforms (coordinator for both CHU Bordeaux and
Bergonié Institute)
• Member of the Regional and Canceropôle Network of Tumour Banks
• Past-Member of the scientific steering committee of the Canceropole Grand Sud-Ouest (2004-2010) (Bordeaux,
Limoges, Montpellier, Toulouse).
• Past-member of several scientific committees in non-governmental cancer research association (ARC, Ligue)
• Member of the French Society of Pathology°
• Member of national groups in the field of clinical research on lymphomas (LYSA) and cutaneous lymphomas
(GFELC)
• Member of the European Association for Haematopathology
SCIENTIFIC AND TEACHING RESPONSIBILITIES
• Head of EA (Research Team) 2406 Histology and Pathology of Tumours. University of Bordeaux.
Rated A in 2010 by National Agency for Evaluation (AERES)
• Coordinator of several training courses in the field of Histology and Biopathology at University of Bordeaux (First
and Second years of medical studies)
• Local Coordinator of the National Training Course in Molecular Pathology (InterUniversity Degree)
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Megan O. Nakashima received an undergraduate degree in Biology from Harvard
University and M.D. from Washington University in St. Louis. She completed Anatomic and
Clinical Pathology residency at the Hospital of the University of Pennsylvania, followed by
fellowship in Surgical Pathology at Barnes-Jewish Hospital/Washington University School
of Medicine. She then trained in Hematopathology at the Cleveland Clinic, and subsequently
remained there as an Associate Staff pathologist in the Division of Hematopathology.
ATTI L I O ORAZI
Dr. Attilio Orazi is Professor of Pathology and Laboratory Medicine, Vice-Chair and
Director of the Division of Hematopathology at the Weill Cornell Medical College (WCMC)
of Cornell University in New York. He serves as Attending Pathologist at WCMC/New
York-Presbyterian Hospital. He received his MD from the University Of Milano School Of
Medicine, in Italy, completed residency training in Clinical Hematology and in Pathology in
the United Kingdom and in Italy. He had previously served as Director of Hematopathology
at the Columbia University Medical Centre in New York and at Indiana University School of
Medicine in Indianapolis, Indiana. He has published more than 200 peer reviewed medical
journal articles and more than 30 book chapters primarily in hematopathology. He is the
lead editor of one of Knowles Neoplastic Hematopathology 3rd Edition. He is lead author or co-author of three additional
textbooks. He is a senior advisor and is actively involved in undergoing updating of the current WHO Classification of
Hematopoietic and Lymphoid Tissues Tumours.
MEGA N O. NAK ASHIM A
C HRI S TOPHER Y. PARK
My group investigates the origins of hematologic malignancies by characterizing the
molecular and cellular changes present in highly-purified patient disease-initiating (i.e.
cancerstem) cells. We also study normal hematopoietic stem cells (HSCs) in order
to understand how dysregulation of normal pathways contribute to disease stemcell
function and disease pathogenesis. Major lines of investigation in the lab include: 1)
microRNA regulation of HSC function; 2) mechanisms of transformation by leukemogenic
microRNAs; 3) identification of novel leukemia stem cell (LSC) markers and validation
of such markers in the clinical setting; 4) functional and molecular characterization
of disease-initiating cells in the myelodysplastic syndromes (MDS), acute myeloid
leukemia (AML), and mature B-cell neoplasms, including hairy cell leukemia (HCL); 5)
identification of therapy resistance genes in AML stem cells. Our long-term goals are to identify novel diagnostic
and prognostic markers and to develop novel disease stem cell-directed therapies.
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PI ER PAOLO PICCALU GA
Associate Professor of Pathology
Department of Experimental, Diagnostic and Specialty Medicine, Bologna University
School of Medicine.
Via Massarenti, 9 - 40138 Bologna - Italy
Tel. +39/051/6364043 - 6363680
Fax +39/051/6364037
e-mail: pierpaolo.piccaluga@unibo.it
Born in Bologna, Italy on February, 5th 1973
1991
1991 to 1997
1997 to 2001
: Classical high school degree.
: Attending classes at the University of Bologna Medical School: Internship in Internal Medicine,
Pathology and Hematology from 1993 to 1997
Degree in Medicine and Surgery in 1997 with honors and “dignità di stampa” with a thesis entitled
“Peripheral T-cell lymphomas. Clinico-pathologic study of 168 cases diagnosed according to the
R.E.A.L. Classification”.
: Fellowship/Residency in Hematology (Specialty degree in Hematology with honors in November 2001
defending a thesis entitled “Neoangiogenesis in the hematologic malignancies”).
January 2002 to
June 2005
: PhD in Clinical and Experimental Hematology (thesis defense in June 2005: Gene expression
analysis of peripheral T-cell lymphoma by DNA and tissue microarrays) .
2003-2004
: Post doctoral research fellowship at the Institute for Cancer Genetics (Columbia University, New
York, NY) .
2008
: Visiting Scientist at the Center for Computational Biology and Bioinformatics, Columbia University
Medical Center.
Core Lab Director – Core Lab Program Affymetrix, Oncopharmagen Group 2008;
2009
: Visiting Scientist at the Institut fur Pathologie, Christian-Albrechts-University, Kiel, Germany.
2006-2012
: Assistant Professor (Lecturer) in Pathology, Unit of Hematopathology, Department of Hematology
and Oncological Sciences “L. & A. Seràgnoli”, University of Bologna;
June 2010
: Obtainment of the title of Associate Professor of Pathology at the University of Perugia
September 2012 to
present
: Associate Professor of Pathology, Department of Experimental, Diagnostic and Specialty
Medicine, Bologna University School of Medicine, Molecular biology laboratory, Hematopathology
Unit.
January 2014 : Obtainment of the “Abilitazione Nazionale a Professore Ordinario, SSD MED/08”
July 2014
: Fellowship in Pathology with honors at the University of Siena
Author of several international publications indexed in the Current Contents (180 publications quoted in PubMed; total
impact facto >1,000; mean impact factor >6 -http://www.ncbi.nlm.nih.gov/sites/entrez; Piccaluga P). H-index: 38 (Rank
89 in the Top Italian Scientist list).
Author of several presentations at national and international conferences.
Involved in several clinical trail as sub-investigator or coordinator.
Winner of several prizes for study and research.
Involved/PI in several research project granted by national and international recognized organisms.
Invited referee for several Journals indexed in the Current Contents.
Associate Editor for the American Journal of Blood Research, Modern Chemotherapy, World Journal of Hematology.
Member of the following societies: SIE, SIES, ASH, AACR, SIAPEC/IAP, EHA, ELN/EWALL, FIL.
Main research interests:
Pathobiology of peripheral T-cell lymphomas
Novel treatments for non Hodgkin lymphomas
Pathobiology of Burkitt lymphoma
Biology and treatment of acute myeloid leukemia
Biology and treatment of acute lymphoblastic leukemia
Myelofibrosis and microenvironment
44
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Born April 30th, 1947. Full-Professor of Pathologic Anatomy, Director of the Service of
Haematopathology, BolognaUniversity.
Over 10,000 diagnoses signed per year since 1980, at present corresponding to haematopathologic biopsies sent from 85 Italian Centres.
Author of 1,048 scientific reports (574 quoted in PubMed and 2 in press in International
Journals with a total IF of 3,403.848) and regular invited speaker at International Meetings.
Co-author of the “Revised European American Lymphoma Classification” (1994). Coeditor and Co-author of the fourth Edition of the WHO Classification of Lymphomas
and Leukaemias. Past-President of the European Association for Haematopathology. H.C. degree in Medicine at
AthensUniversity. Ranked 42nd among the Italian Scientists of the last Century because of his H-index (=92).
X OSE S . PU ENT E
Dr. Xose S. Puente is a Professor of Biochemistry and Molecular Biology at the
University of Oviedo (Spain). He has been working on the molecular basis of cancer for
more than 25 years, as well as the sequencing and analysis of genomes from different
model organisms to understand human physiology. During the last five years he has
been working on the genomic analysis of chronic lymphocytic leukemia and mantle cell
lymphoma, in which more than 500 tumor samples are being sequenced, developing
novel analysis tools and techniques for the rapid analysis of genes using next generation
sequencing technologies. His work has allowed the identification of more than a dozen
novel oncogenes mutated in this hematological neoplasia, many of them with clinical
impact on the evolution of the disease, paving the way for the introduction of genetic
classification in the management of cancer patients.
S TE FA NO A. PILERI
BENJA M IN RENGSTL
Benjamin Rengstl studied biochemistry and medicine at the Goethe-University Frankfurt,
Germany. After graduating in biochemistry, he received a student award for his diploma
thesis on antigen presentation performed at the Federal Institute for Vaccines and
Biomedicines (Paul-Ehrlich-Institute) in Langen, Germany. This year he earned the PhD
degree from the Goethe-University Frankfurt and parts of his thesis entitled “On the
origin of giant Reed-Sternberg cells and the role of CD4 T cells in Hodgkin lymphoma”
have been published in PNAS and honored with the research award of the German
Society of Pathology. Besides completing the final clinical year at the medical school,
he currently holds a postdoctoral fellowship of the lymphoma research group located
in the Dr. Senckenberg Institute of Pathology in Frankfurt, Germany. The main interest
of research and topic of his MD thesis is now focusing on the interactions between lymphoma cells and the
immunological microenvironment.
L I S A RI M SZA
Lisa Rimsza, M.D., is a tenured full Professor of Pathology and Associate Chair of
Research at the University of Arizona, Tucson, Arizona, U.S.A. She is a practicing
Hematopathologist and directs the Molecular Genetic Pathology fellowship. She is a
member of the Southwest Oncology Group Lymphoma Pathology and Translational
Medicine Committees, International Lymphoma Study Group, and chairs the National
Institutes of Health Lymphoma Biomarker Committee. Currently, she is the Principal
Investigator of an international research consortium, the Lymphoma & Leukemia Molecular
Profiling Project, which uses gene expression profiling for lymphoma diagnosis and
prognosis. She also holds awards from the National Cancer Institute for the Specialized
Center of Research Excellence in Lymphoma, Tissue Acquisition and Molecular Analysis
Shared Resource at the Arizona Cancer Center, and AIDS Cancer Specimen Research Network. Her own laboratory
focuses on mechanisms of tumor immunosurveillance and therapy resistance, including detection and targeting of the
BCL2 and MYC oncogenes.
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RI C HA RD ROSENQU IST
Richard Rosenquist is professor of Molecular Hematology at the Department of
Immunology, Genetics and Pathology, Uppsala University, and head of the Cancer
Genetic Section at Uppsala University Hospital, Sweden. He has longstanding
experience in development, clinical validation and standardization of novel biomarkers/
technologies in hematology with a particular focus on CLL. Professor Rosenquist is also
Platform Director for the newly established Clinical Sequencing Facility within Science
for Life Laboratory, Uppsala (www.scilifelab.se) with the aim to translate next-generation
sequencing into clinical diagnostics.
RUSSELL J.H. R YAN
I am a graduate of the Massachusetts General Hospital training programs in Anatomic
Pathology and Hematopathology, and am currently pursuing a research fellowship in the
laboratory of Dr. Bradley Bernstein at MGH and the Broad Institute. My work is focused
on the interaction between the genetic and epigenetic aberrations that drive B cell
lymphoma subtypes, including diffuse large B cell lymphoma, follicular lymphoma, and
mantle cell lymphoma. Specific subtypes of lymphoma show unique patterns of mutations
in genes encoding chromatin regulatory enzymes (CR’s), such as EZH2, as well as CRrecruiting transcription factors. Through computational analysis of genome-wide chromatin
modifications, transcription factor binding, and gene transcription in cell lines and primary
lymphomas, we seek to understand how these mutations ultimately lead to aberrant gene regulation. One goal of this
effort is to improve our understanding of uniquely dysregulated pathways in lymphoma subtypes, leading to novel
subtype-specific therapies and diagnostic biomarkers.
EL ENA SAB AT T INI
Born in Bologna 11/08/1963
Graduation in Medicine with highest vote (voti 110/110 cum laude) at Bologna University
(March 1989).
Surgical Pathology Specialization at Siena University (July 1994) with highest vote (70/70).
WORKING CARRIER
Fellowship at the Haemolymphopathology Unit in Bologna University (director Prof. Stefano
Pileri) with stable position as haematopathologist (1993).
Referent for Quality Assurance for the Haematopathology Unit since 2002.
Responsible for the High Specialty Program “Lymphoproliferative Disorders” (evaluation
every three years)
Member of the European Association for Haematopathology since 1990.
Member of the Executive Committe of the European Association for Haematopathology for the period 2010-2014.
TEACHING
Tutorage for fellows, residents and guests attending the Unit.
Temporary-teacher at Bologna University for 1) Hematology, Pathology and School Biomedical Techiniques Schools
of Specialization of Bologna University (Biotecnnologies in haematopathology; Bone marrow Pathology, Special
Histopathology)
PUBLICATIONS
Link to the US National Lbrary of Medicine National Institute of Health:
http://www.ncbi.nlm.nih.gov/pubmed/?term=Sabattini+E
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Gilles Salles is a Professor at the University Claude Bernard de Lyon and Head of the the
Department of Haematology of the Hospices Civils de Lyon, at the Centre Hospitalier LyonSud, Lyon, France. He is also the head of the Research Unit ‘Indolent B cell Proliferations”’
at the University of Lyon and affiliated with the CNRS. He is the first chairman of lymphoma
cooperative group LYSA (The Lymphoma Study Association) since 2012. He also serves
as vice-president for research in the board of Directors of the Lyon University Hospitals
(Hospices Civils de Lyon). Professor Salles has been especially interested in the clinical and
biological study of malignant lymphoma – major focuses of his work include the description
and validation of prognostic factors as well as clinical trials in indolent lymphomas. He has
been involved as a coordinator or co-investigator in many clinical trials and studies within
his field, and has published numerous articles in international peer-reviewed journals.
MA RG ARITA SANCHEZ-B EAT O
Margarita Sanchez-Beato was born in Toledo (Spain) and she graduated in Chemistry
(Biochemistry and Molecular Biology) in 1990 at Universidad Complutense (Madrid). In
1992 she joined the Departments of Pathology and Genetics in Hospital “Virgen de la
Salud” in Toledo and obtained her PhD degree in Chemistry (by Universidad Complutense)
in 1997 under the supervision of Miguel A. Piris.
In 2000 she joined the Molecular Pathology Programme at Spanish National Cancer
Research Centre (CNIO), where her work focused on deciphering the genetic and
epigenetic alterations of lymphomas.
Margarita moved to the Institute of Research “Puerta de Hierro” (IDIPHIM) in Madrid in
2012 as Head of the Group of Research in Lymphomas, where her work continues to focus
on the identification of molecular predictor markers and therapeutic targets in lymphomas.
She has authored more than 65 articles and has made important contributions to the field of molecular characterization
of lymphomas.
G I L L E S SALLES
JA NI NE -ALISON SCHM IDT
Janine-Alison Schmidt was born in Tübingen, 1986. After finishing school she came back
to Tübingen in 2005 to begin her studies of Biology which she completed in 2011 writing
her Diploma thesis on the topic of surface marker expression in anaplastic large cell
lymphoma in the working group of Prof. Leticia Quintanilla-Fend and Prof. Falko Fend. In
the same year she started as a PhD student in the same group focusing now on follicular
lymphoma (FL) and its early precursor form, follicular lymphoma in situ (FLIS). Here she
works on the identification of early secondary alterations in FLIS that might represent
drivers for the progression to FL and first results were published in Leukemia in 2013.
Furthermore she is working on lymphoplasmacytic lymphoma (LPL) and developed a
method to analyze MYD88 L265P mutations in FFPE bone marrow tissues and also
investigated the correlations of CXCR4 mutations and clinical data in LPL.
RI TA S HAK NOVICH
Rita Shaknovich, M.D., Ph.D. is a physician scientist with strong interests in cancer biology
and translational research. She received her MD/PhD degrees form Mount Sinai School
of Medicine (NY, USA), completed her Residency in Pathology in Columbia Presbyterian
(NY) and fellowship in Hematopathology in Cornell Medical Center (NY). She is currently
as Assistant Professor in the Department of Pathology and Medicine in WCMC (NY).
Her clinical expertise is in Hematopathology with a focus on lymphoid malignancies. Her
laboratory studies epigenetics of normal B cell development and lymphomagenesis; role
of DNA methylation machinery in clonal diversification of B cells during the germinal center
transit and its contribution to transcriptional deregulation in lymphomas with particular
focus on DLBCLs and CLL. They successfully utilize many experimental and computational tools that allow for the
genome-wide profiling of DNA methylation and gene expression, like ERRBS, CHIP-seq, RNA-seq, exome and whole
genome sequencing among others.
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I MRA N N. SIDDIQI
Dr. Imran N. Siddiqi is Assistant Professor of Clinical Pathology at the Keck School of
Medicine of the University of Southern California in Los Angeles. He was Assistant Professor
at the University of California San Francisco prior to joining the USC faculty in 2010. He
currently serves as Program Director of the Hematopathology Fellowship Program and
is the Medical Director of the Hematology Laboratories at USC Norris Cancer Hospital
and Keck Hospital of USC. Dr. Siddiqi completed his residency training at UCSF with
fellowships in Surgical Pathology and Hematopathology. He is board certified in Anatomic/
Clinical Pathology and Hematology by the American Board of Pathology. Dr. Siddiqi’s
research interests include immunohistochemical and molecular biomarkers for diagnosis
and prognosis of hematolymphoid malignancies. He has authored over 20 papers in peerreviewed journals as well as several book chapters and has presented his work at numerous conferences. Dr. Siddiqi’s
areas of diagnostic expertise include benign and malignant diseases of the lymph nodes, extranodal lymphoid tissues,
bone marrow and blood.
A NNETTE M . STAIGER
PhD study:
Date of Birth:
15.09.1971
Born in:
Stuttgart
Study:
2002 – 2010 Technical Biology at the University of Stuttgart, graduated
with Master of Science
Master thesis
“Characterization and pharmacological modulation of oncogenic
stress induced by p190Bcr-Abl-overexpression” in the working group
of Professor Dr. Aulitzky, Margarete Fischer-Bosch-Institute of Clinical
Pharmacology (IKP), Stuttgart, Germany
PhD student and member of the working group of Professor Dr. Ott at the IKP Stuttgart since
2011. Focus of the thesis is on gene-expression based prognostic models in follicular lymphoma,
supervised by Dr. Heike Horn.
Responsibilities: Since 2013 coordination of the molecular diagnostics Unit of the department of Clinical Pathology,
Head Prof. Ott, Robert Bosch Hospital, Stuttgart.
L OUI S M.STAU DT
Dr. Staudt received his B.A. from Harvard College in 1976, graduating Cum Laude in
Biochemistry. He was awarded a Medical Scientist Training Program fellowship at the
University of Pennsylvania School of Medicine and received his M.D. and Ph.D. degrees
in 1982. His Ph.D. thesis in the field of immunology, performed in the laboratory of Walter
Gerhard, revealed somatic hypermutation as a mechanism of rapid antibody diversification
during normal immune responses. Following Internal Medicine training, he joined Nobel
Laureate David Baltimore’s laboratory at the Whitehead Institute as a Jane Coffin Childs
Fellow. There he cloned and characterized the first tissue specific transcription factor, Oct2. He established his laboratory in the Metabolism Branch, National Cancer Institute (NCI)
in 1988 and is currently Co-Chief of the NCI Lymphoid Malignancies Branch. He is also
Director of the NCI Center for Cancer Genomics, which oversees several large-scale managed programs studying the
genomic aberrations in cancer. In 2011, Dr. Staudt was given the honorary title of NIH Distinguished Investigator. Dr.
Staudt serves on the Editorial Boards of Cancer Cell, the Journal of Experimental Medicine and eLife. He has received
numerous awards for his research, including the 2009 Dameshek Prize from the American Society of Hematology
for outstanding contribution in hematology and elected to the National Academy of Sciences in 2013. Dr. Staudt’s
laboratory uses genomics to improve diagnosis and treatment of lymphomas.
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Michael R. Stratton is Director of the Wellcome Trust Sanger Institute. His primary research
interests have been in the genetics of cancer. His early research focused on inherited
susceptibility. He mapped and identified the major high risk breast cancer susceptibility
gene BRCA2 and subsequently a series of moderate risk breast cancer and other cancer
susceptibility genes.
In 2000 he initiated the Cancer Genome Project at the Wellcome Trust Sanger Institute
which conducts systematic genome-wide searches for somatic mutations in human cancer.
Through these studies he discovered somatic mutations of the BRAF gene in malignant
melanoma and several other mutated cancer genes in lung, renal, breast and other cancers.
He has described the basic patterns of somatic mutation in cancer genomes revealing
underlying DNA mutational and repair processes.
He is a Fellow of the Royal Society (FRS) and was Knighted by the Queen in 2013.
VA NESSA SZAB LEWSK I
Dr Vanessa Szablewski studied Medicine in Montpellier where she specialized in pathology.
She worked for 6 month with Professor Philippe Gaulard and Professor Christiane Copie
in Henri Mondor Hospital, Créteil, where she acquired experiment on haematopathology.
Currently she has in charge the haematopathology activity and the FISH (Fluorescent In
Situ Hydridization) activity on solid and hematopoietic tumor in the Department of Pathology
of Montpellier.
She continues collaborative projects with Professor Philippe Gaulard and Professor
Christiane Copie, in particular on Follicular Lymphoma.
Her future goal is to set up an hemato-oncogenetic platform on the site of CHU de Montpellier where clinicians,
biologists and pathologists could work together and bring their own competences.
MI CHA EL R. STRATTON
P R L UC XER R I
Pr Luc XERRI, PhD, MD, is Professor of pathology at the “Aix-Marseille » University of
Marseilles, France since 1999. Head of the department of bio-pathology of the Institut
Paoli-Calmettes (center against cancer of Marseilles) since 2002. Member of the pathology
team of the former “Groupe d’Etude des Lymphomes de l’Adulte” (GELA), recently renamed as the “Lymphoma Study association” (LYSA). He is in charge of the pathological
reviewing process in follicular lymphoma clinical trials, including the FL2000, PRIMA and
RELEVANCE Trials. Member of the French national network of experts for lymphoma
diagnosis (LYMPHOPATH). His research fields include co-signaling receptors and immune
microenvironment in lymphoma pathogenesis, and the identification of bio-pathological
prognostic markers in Hodgkin’s lymphoma and follicular lymphoma.
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ORAL PRESENTATIONS
Jon C. Aster1, Ami Bhatt2, Birgit Knoechel2, Daniel J. Deangelo2,
Li Pan1, Michael J. Kluk1, Frank Kuo1, Matthew Meyerson2
1
2
Department of Pathology, Brigham and Women’s Hospital, Boston, MA USA
Department of Medical Oncology, Dana Farber Cancer Institute, Boston, MA USA
T-cell lymphoblastic leukemia/lymphoma (T-LL) is associated with Notch1 gain-offunction mutations in roughly 50% of cases. The highest fraction of Notch1-mutated
cases corresponds to T-LL with a cortical immunophenotype, but recent work has
suggested that up to 50% of patients with adult early T progenitor lymphoblastic
leukemia (ETP-LL), a subtype of T-LL corresponding to early stages of T cell commitment, are also Notch1 mutated. Most previous clinical trials of gamma-secretase
inhibitors (GSIs), drugs that inhibit Notch activation, have produced at best partial
responses in patients with relapsed refractory T-LL, most or all of which have been
of cortical subtype. Here we report a middle age male with refractory ETP-LL who
achieved a complete hematologic remission with a GSI being tested in a phase
I trial. Short-term culture confirmed the leukemic blasts contained high levels of
activated Notch1 at baseline that were rapidly depleted by incubation with GSI.
To understand the molecular basis of response in this sentinel case, whole exome
sequencing was performed on DNA prepared from leukemic blasts and remission
marrow. The leukemic blasts contained four pathogenic driver mutations: i) a heterozygous Notch1 F1592C gain-of-function mutation; ii) a homozygous/hemizygous
DNMT3a Q402* loss-of-function mutation; iii) a heterozygous Shp2 F285S gain-offunction mutation; and iv) a heterozygous T1681T CSF3R gain-of-function mutation. Resequencing of DNA from remission marrow revealed wildtype Notch1, Shp2,
and CSF3R sequences and a heterozygous loss-of-function mutation in DNMT3a,
whereas sequencing of DNA obtained saliva showed only wildtype sequences for
DNMT3a. From these results we deduced that this ETP-LL developed from a preexistent stem cell clone with an initiating heterozygous DNMT3a mutation via acquisition of additional mutations in Notch1, Shp2, and CSF3R and loss of the remaining
normal copy of DNMT3a. To the best of our knowledge, this case is the first example of human ETP-LL arising from an identifiable precursor state, and also serves
to show that Notch-“addiction” can be acquired even in tumors in which Notch1
mutations are secondary, rather than initiating, events. Work is now underway to
identify Notch1 target genes in “super-responder” lymphoblasts and to study tumor
response to GSI +/- other targeted therapies in immunodeficient mice. It is hoped
that understanding the basis of GSI response in this sentinel case will lead to the
identification of biomarkers that reliably predict GSI responsiveness and resistance
in relapsed/refractory T-LL.
Keywords: Notch, leukemia, targeted therapy
We validated our findings on 33 biopsies from 30 patients using immunohistochemistry. Cases were divided into 4 groups based on morphology and Ki-67 staining in
order to reflect the phenotype associated with disease aggressiveness: typical CLL
(n=12, G1), CLL with expanded Proliferation Centers (n=12, G2), CLL with increased
number of Large Cells (n=6, G3) and Richter’s Transformaiton (n=3, G4). Based on
immunohistochemical stains, more aggressive G3 had significanly higher Ki-67, p53
and c-MYC, compared to G1 and G2 supporting methylation findings.
Conclusions: We identified novel epigenetic subgroups of CLL and associated aberrant epigenetic events and validated upregulation of C-MYC in cases with more
aggressive morphologic features.
ORA L P RES EN TAT I ON S
COMPLETE RESPONSE OF ETP-LL TO A NOTCH PATHWAY
INHIBITOR: MOLECULAR AND CELLULAR CHARACTERIZATION
OF A SENTINEL CASE
profiling would allow us to identify new, biologically significant CLL subtypes and
yield greater insight into biology of this disease. We therefore examined the DNA
methylation of over 240 patients with CLL using the HELP assay and hybridization to high density custom microarray that reports on methylation status of more
than 250,000 CpGs corresponding to 20,000 genes. Gene expression profiling and
SNP arrays and targeted resequencing were available on most of these cases. We
next performed an unsupervised analysis of probesets that display significant variability using three statistical approaches: K-means consensus clustering, Pearson
correlation and Principal component analysis. This procedure reproducibly identified
three robust CLL subtypes based on epigenetic profiles. To identify the genes that
define these three subtypes we next performed unequal variance t-test of the CLL
subtypes comparing them to Peripheral Blood CD19+ B cells as a normal control
and identified that clusters are defined by differential methylation 4112, 6364 and
3771 genes respectively (selected probes displayed change in methylation of at
least 30% at FDR corrected p-value < 0.05), Unfavorable outcome cluster captured
cases with known biomarkers predictive of unfavorable clinical outcome (CD38+,
ZAP70+ and IgVH unmutated). Integration of methylome and transcriptome results
followed by pathway analysis using GSEA, iPAGE and IPA revealed that favorable
outcome cluster was characterized by deregulation of B cell differentiation program;
intermediate outcome cluster was characterize by activation of DNA damage repair
pathways including KAT5 and unfavorable outcome cluster was characterized by
activation of NF-kB signaling and disruption of MYC/WNT signaling.
[OP-BM-01]
Keywords: Chronic Lymphocytic Leukemia, epigenetics, DNA methylation
[OP-BM-03]
NON-IGM LYMPHOPLASMACYTIC LYMPHOMA SHOWS
PATHOLOGIC AND CLINICAL HETEROGENEITY AND MAY
HARBOR MYD88 MUTATIONS
Rebecca L. King1, Wilson I. Gonsalves2, Matthew T. Howard1,
Lori A. Frederick1, David S. Viswanatha1, Patricia T. Greipp3,
Stephen M. Ansell2, William G. Morice1
1
Division of Hematopathology, Mayo Clinic, Rochester, MN, USA
Department of Hematology/Oncology, Mayo Clinic, Rochester, MN, USA
3
Division of Laboratory Genetics, Mayo Clinic, Rochester, MN, USA
2
[OP-BM-02]
EPIGENETIC PROFILING OF CLL REVEALS NOVEL DNA
METHYLATION-BASED CLUSTERS AND NOVEL MECHANISMS
OF LYMPHOMAGENESIS
Fang Fang1, Julia Geyer2, Wayne Tam2, Richard R. Furman1,
Yi Fang Liu2, Peter Ouillette3, Erlene Seymour3, Kamlai Saiya Cork3,
Kerby Shedden4, Daniel M. Knowles2, Ari M. Melnick1,
Sami N. Malek3, Attilio Orazi2, Rita Shaknovich2
1
Division of Hematology and Oncology, Weill Cornell Medical College, New York, USA
Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, USA
3
Department of Internal Medicine, Division of Hematology and Oncology, University of Michigan, Ann
Arbor, USA
4
Department of Statistics, University of Michigan, Ann Arbor, MI, USA
2
Chronic Lymphocytic Leukemia (CLL) is a common B cell malignancy in the Western
world with more than 15,000 cases diagnosed annually. Attempts to characterize
CLL resulted in identification of disease biomarkers: recurrent genomic deletions
del11q and del17p along with overall genomic complexity; expression level of
CD38 and ZAP70; mutational status of Ig heavy chain genes. While genomic and
transcriptional profiling of CLL identified clinically and biologically relevant markers, there is still significant uncertainty about the pathobiology and the origin of
CLL. It is increasingly clear that epigenetic deregulation plays an important role in
the biology of all lymphomas including CLL. We hypothesized that DNA methylation
Waldenstrom Macroglobulinemia (WM) is a clinical syndrome defined as lymphoplasmacytic lymphoma (LPL) with an associated IgM serum paraprotein. Recently,
mutations in the MYD88 gene have been identified in approximately 90% of WM
cases. MYD88 mutation testing presents a valuable diagnostic aid, as the pathologic
features of LPL bone marrow infiltrates overlap with those of other low grade B
cell lymphomas. Although the clinical diagnosis of WM is restricted to cases with
IgM paraproteins, the WHO recognizes that LPL can rarely present with paraproteins of other heavy chains. However, due to the paucity of literature describing
the clinicopathologic features of LPL associated with non-IgM paraproteinemia, the
relationship of such cases to classical WM remains unclear. As such, the goals of
our study were to identify such LPL cases and evaluate their clinical, pathologic,
and genetic features.
Twenty three bone marrow cases diagnosed as either lymphoplasmacytic lymphoma or low grade B cell lymphoma with plasmacytic differentiation that had either
IgG or IgA paraproteins were identified in the Mayo Clinic pathology archives from
2007-2013. None of the cases had an extramedullary tissue diagnosis of marginal
zone lymphoma or other non-LPL low grade B cell lymphoma.
The degree of marrow involvement ranged from 5 to 90% (median 20%).
Morphologically, 18 (78%) cases showed the typical spectrum or mixture of lymphoid and plasmacytic cells, while 5 were described as having predominantly either
small lymphocytes or plasma cells. Plasma cells had Dutcher bodies in 5 cases.
Flow cytometry detected both clonal B cells and clonal plasma cells in 18/20 (90%)
cases in which this testing was performed. Plasma cells showed complete loss of
CD19 in 6 (35%) cases and at least partial positivity in 11 cases (remaining one case
| 17-22 October 2014
| EAHP - 2014
|
53
In conclusion, non-IgM LPL involving the bone marrow shows heterogeneous
morphologic and immunophenotypic features that are indistinguishable from IgM
LPL. Furthermore, MYD88 mutations are present in a substantive proportion of
cases, although less than expected based on the published frequency in IgM associated LPL. While most cases do not have the hyperviscosity prototypic of classic
WM, some patients meet the recognized symptomatic criteria for a WM diagnosis
and may respond favorably to WM-type therapy. The clinical symptoms of WM in
this group did not correlate with MYD88 mutations status. Overall, these findings
suggest that at least a subset of LPL associated with non-IgM paraprotein are
clinically and biologically similar to classic WM although additional phenotypic
and genetic studies on these cases are warranted to further refine this diagnostic
entity.
Keywords: lymphoplasmacytic lymphoma, waldenstrom, non-IgM paraprotein
Table 1. Clinical features of non-IgM lymphoplasmacytic lymphoma
th
not tested for CD19). The plasma cells were CD45 negative in 3/18 (17%) cases.
MYD88 L265P mutations were present in 6/23 (26%) cases. Table 1 highlights the
spectrum of clinical features typically associated with WM and their occurrence in
these cases.
[OP-BM-04]
erythroid colony forming capacity; treatment of Mx1-Cre BRafV600E mice improved
anemia and splenomegaly.
HEMATOPOIETIC STEM CELL ORIGIN OF HAIRY CELL LEUKEMIA
1
1
1
1
Stephen S. Chung , Eunhee Kim , Jae H. Park , Young Rock Chung ,
Julie Feldstein1, Wenhuo Hu1, Michael F. Berger1, Ari M. Melnick2,
Neal Rosen1, Martin S. Tallman1, Omar Abdel Wahab1,
Christopher Y. Park1
1
2
Memorial Sloan Kettering Cancer Center, New York, NY, USA
Weill Cornell Medical College, New York, NY, USA
We examined hematopoietic stem and progenitor cells (HSPC) in the bone marrow
(BM) of HCL patients and found markedly increased hematopoietic stem cells (HSCs)
and decreased numbers of granulocyte-macrophage progenitors. In addition, we
identified the BRAFV600E mutation not only in mature HCL cells, but also in purified
HSCs and pro-B cells. Transplant of HCL patient HSCs into NOD/SCID/IL2-R null mice
led to engraftment of expanded pro-B cells as well as cells expressing HCL cell
markers (CD11c+CD103+CD19+). These engrafted cells harbored the BRAFV600E
mutation, strongly suggesting that the disease-initiating cell in HCL is the HSC.
Conditional expression of BRafV600E from its endogenous locus at two different
stages of hematopoiesis was achieved by crossing BRafV600E mice with Mx1-Cre
and Cd19-Cre mice. Mx1-Cre BRafV600E mice developed a lethal hematopoietic
malignancy characterized by features of HCL including splenomegaly, anemia,
thrombocytopenia, increased circulating sCD25, and increased clonogenic capacity
of B-lineage cells (with infinite serial replating capacity in the presence of IL-7).
Cd19-Cre BRafV600E mice demonstrated the same increased clonogenic capacity
of B-lineage cells, but exhibited no other phenotype up to one year of age. These
data suggest that HCL-associated cytopenias are due to cell-intrinsic defects in
BRAFV600E mutant HSPCs. Treatment of HCL patients with the BRAF inhibitor
Vemurafenib led to normalization of HSPC frequencies and restoration of myelo/
| EAHP - 2014
| 17-22 October 2014
Keywords: Hairy cell leukemia, hematopoietic stem cell, cancer stem cell
[OP-LYMP-01]
Hairy cell leukemia (HCL) is a chronic lymphoproliferative disorder characterized
by pancytopenia, splenomegaly, and somatic BRAFV600E mutations. The malignant
cell in HCL is a mature B cell with surface expression of the pan-B-cell marker CD19
and clonal rearrangements of immunoglobulin heavy and light chains. However, HCL
cells also express non-B lineage associated markers such as CD11c and CD103,
and we thus hypothesized that HCL may originate from a less mature progenitor.
54
Together, these findings link the pathogenesis of HCL to somatic mutations that
arise in HSPCs and support a model in which chronic lymphoid malignancies initiate
from aberrant hematopoietic stem cells.
IDENTIFICATION OF SINGLE-NUCLEOTIDE VARIANTS BY RNA
SEQUENCING IN ENDEMIC BURKITT LYMPHOMA
Pier Paolo Piccaluga1, Francesco Abate2,
Maria Antonella Laginestra1, Giulia De Falco3, Maryam Etebari1,
Fabio Fuligni1, Maura Rossi1, Sakellarios Zairis2, Cristiana Bellan3,
Lorenzo Leoncini3, Raul Rabadan2, Stefano Aldo Pileri1
1
Department of Experimental, Diagnostic, and Specialty Medicine, University of Bologna, Bologna,
Italy
2
Columbia University College of Physicians and Surgeons, New York, NY-USA
3
Department of Medical Biotechnologies, University of Siena, Siena, Italy
Burkitt lymphoma (BL) is an aggressive B-cell malignancy comprising three clinical
variants named endemic (eBL), sporadic (sBL) and immunodeficiency associated
(ID-BL). Endemic Burkitt lymphoma (eBL) is the commonest cancer in children in
Developing Countries, while the other two forms are mainly encountered in Western
Countries. The molecular hallmark of BL is the translocation of the oncogene MYC to
the immunoglobulin-heavy [t(8;14)(q24;q32)] or one of the light chain genes [t(2;8)
(p12; q24) and t(8;22)(q24; q11)], leading to constitutive MYC expression. However,
additional genetic events contributes to BL pathogenesis, as recently shown in sBL.
We performed global RNA-Sequencing (Illumina HiScanSQ) of eBL aiming to identify
genetic changes possibly cooperating with MYC aberrant expression in the pathogenesis of the disease. We studied 21 eBL cases, collected at different African
Institutions as discovery set. Total RNA was extracted with Trizol and libraries were
prepared according to TruSeq RNA sample preparation v2 protocol. Sequence variants were obtained using the SAVI (Statistical Algorithm for Variant Identification)
Sanger sequencing confirmed such SNVs with 100% accuracy within the discovery set. therefore, we sought to evaluate their frequency in a screening set of
cases (N=78) by both Sanger sequencing and mass spectrometry (Sequenom). We
found that most eBL cases carried additional SNVs rather than MYC translocations.
Noteworthy, significant differences were found in eBL and sBL concerning the mutation rate in different genes, including among others ID3, TCF3, ARID1A, CCNF, and
HERC2, sustaining the hypothesis that different pathogenetic events may contribute
to the pathogenesis of BL subtypes.
Conclusions: This is the first thorough biological and pre-clinical dissection of: i)
the prominent role of BRAF-V600E in shaping the specific transcriptional signature,
morphology and immunophenotype of HCL; and ii) the significant anti-leukemic activity of BRAF or MEK inhibition in HCL.
Keywords: Hairy Cell Leukemia, BRAF-MEK-ERK pathway, Targeted therapy
In conclusion, we discovered new SNVs that might have a significant role in the
pathogenesis of eBL. Functional experiments are required to definitely assess their
impact.
Keywords: Burkitt lymphoma, RNA-sequencing, single nucleotide variant
We found 66 genes affected by 219 total SNVs with different frequency in 21 samples (range 2-22 SNV/sample). We then focused on genes recurrently mutated (in
at least 3/21 samples) and for which somatic protein-changes were predicted. We
identified 25 genes affected by 172 total SNVs, that were recurrently mutated (min.
3/21 samples; max 11/21 samples). These included genes previously known to be
involved in sBL, such as TP53, MYC, ID3, PCBP1 and TCF3 as well as genes involved
in other lymphomas such as DDX3X and RHOA. The remaining 18 genes (AGAP6,
APBB1IP, ARID1A, ASPSCR1, AVEN, CAD, CCNF, GPATCH4, HERC2, KPNA2, MTBP,
MTERFD1, NEK9, NUP133, PARP1, POLQ, SCFD2, and TIGD1) were previously not
related to BL, and resulted to be involved in several important molecular pathways
as cell cycle progression, apoptosis, matrix remodeling, and angiogenesis.
downregulation of the HCL immunophenotypical markers CD25, TRAP and cyclin-D1.
Loss of surface CD25 and of nuclear cyclin-D1 was validated in vivo in HCL patients
being treated with Vemurafenib in our HCL-PG1 phase-2 clinical trial. These in vitro biochemical and transcriptional events were followed by a consistent: i) loss of
the hairy projections in still viable (AnnexinV-negative) leukemic cells (see Figure);
ii) reduction of metabolic viability (up to 51.7% relative to the drug vehichle); and iii)
decrease of living (AnnexinV-negative) cells (up to 84.4% relative reduction), all of this
occurring specifically in leukemic cells of the vast majority of HCL patients as opposed
to none of the HCL-like patients. Furthermore, to recapitulate in vitro the physiologic
microenvironment of HCL cells in vivo, and to investigate its potential modulating effect on BRAF inhibitors’ activity, we cultured HCL cells with or without the bone marrow stromal cell line HS5. The latter partially blunted drug-induced MEK/ERK dephosphorylation and apoptosis induction, with Dabrafenib apparently less affected than
Vemurafenib by such protective stromal effect.
algorithm independently for each sample. Candidate somatic mutations were obtained by eliminating common germline variants and recurrence. To validate singlenucleotide variants (SNVs) we performed Sanger sequencing on these 21 cases.
Further, we studied these SNVs by Sequenom Technology and Sanger sequencing
on additional 78 eBL cases as screening set. Twenty-eight sBL cases, for which
RNA-sequencing data were already available, were considered as controls.
[OP-LYMP-02]
BIOLOGICAL AND THERAPEUTIC RELEVANCE OF THE BRAFMEK-ERK PATHWAY IN HAIRY CELL LEUKEMIA
Valentina Pettirossi1, Alessia Santi1, Elisa Imperi1, Guido Russo1,
Alessandra Pucciarini1, Barbara Bigerna1, Elisabetta Fortini1,
Roberta Mannucci2, Gianluca Schiavoni1, Ildo Nicoletti2,
Maria Paola Martelli1, Ludger Klein Hitpass3, Brunangelo Falini1,
Enrico Tiacci1
1
Institute of Internal Medicine and Oncologic Sciences, University of Perugia, Perugia, Italy
Biochip Laboratory, Institute for Cell Biology - Tumor Research, University of Duisburg-Essen
Medical School, Essen, Germany
2
3
Background: Hairy cell leukemia (HCL) has unique clinico-pathological features.
The BRAF-V600E activating kinase mutation defines HCL among other B-cell lymphomas (Tiacci et al, New Engl J Med 2011;364:2305), including the HCL-like mimics splenic marginal zone lymphoma and HCL-variant. The BRAF-MEK-ERK pathway
thus emerges as an ideal candidate to illuminate the peculiar biology of HCL and as
a therapeutic target to be attacked with clinically available BRAF and MEK inhibitors. However, a comprehensive functional dissection of this pathway in HCL has not
been conducted so far.
Aim: To analyze in vitro the effects of BRAF and MEK inhibition in HCL using patients’
hairy cells, since the putative “HCL” cell lines lack BRAF-V600E questioning their
true HCL origin (Tiacci et al, Blood 2012;119:5332).
Methods: Blood leukemic cells, purified from 23 HCL and 10 HCL-like patients using CD19-MACS, were treated in vitro with a BRAF inhibitor (Vemurafenib, PLX4720
or Dabrafenib) or the MEK inhibitor Trametinib at concentrations up to 1 μM for up
to 96 hours, and monitored for: i) MEK/ERK phosphorylation by Western blotting; ii)
downstream transcriptional changes by genome-wide expression profiling (GEP); iii)
surface morphology changes by confocal microscopy for phalloidin and AnnexinV to
highlight the F-actin-rich hairy projections in still living cells; iv) viability (by MTT or
WST metabolic assays) and apoptosis (by AnnexinV flow cytometry).
Results: Treatment with any BRAF inhibitors resulted in dose-dependent, sustained
MEK and ERK dephosphorylation in all HCL cases, as opposed to vehicle-treated HCL
cells and to inhibitor-treated HCL-like cells. Also Trametinib strongly dephosphorylated
ERK in HCL cells. Interestingly, GEP after 48h and 72h of BRAF inhibition showed: i)
silencing of the BRAF-MEK-ERK pathway transcriptional output; ii) loss of the HCLspecific GEP signature we previously identified (J Exp Med 2004;199:59); and iii)
Figure 1. Vemurafenib-induced hair loss in hcl (but not hcl-like) cells
[OP-LYMP-03]
DIFFUSE LARGE B-CELL LYMPHOMA WITH HIGH
EXPRESSION OF MYC AND BCL2 SHOWS EVIDENCE OF
ACTIVE B-CELL RECEPTOR SIGNALING BY QUANTITATIVE
IMMUNOFLUORESCENCE
Agata M. Bogusz1, Alexandra E. Kovach2, Long P. Le2,
Richard H.G. Baxter3, Aliyah R. Sohani2
1
University of Pennsylvania, Department of Pathology and Laboratory Medicine, Philadelphia, PA, USA
Massachusetts General Hospital, Department of Pathology, Boston, MA, USA
Yale University, Department of Chemistry, New Haven, CT, USA
2
3
Diffuse large B-cell lymphoma (DLBCL) is the most common type of non-Hodgkin
lymphoma, with a poor prognosis in approximately 50% of patients. DLBCL is heterogeneous in its biology and shows variable response to conventional chemotherapy and rituximab. A variety of signaling pathways are involved in the pathogenesis
of DLBCL, including the B-cell receptor (BCR) signaling pathway. We have previously
identified a robust BCR signaling signature on formalin-fixed paraffin-embedded
(FFPE) specimens based on quantitative immunofluorescence (qIF) of phosphorylated proximal BCR-associated signaling molecules: kinases LYN, SYK and BTK
(<pLYN>, <pSYK,pBTK> scores) and cytoplasmic localization of transcription factor FOXO1 (Fcyt) (Bogusz et al., Clin Cancer Res 2012; 18(22):6122-35). Our data
revealed active BCR signaling in almost 50% of patient tumors and implicated qIF
as a tool to identify patients that would benefit from anti-BCR therapies such as
fostamatinib or ibrutinib.
Concurrent high expression of MYC and BCL2 by immunohistochemistry (doubleexpressing lymphomas or “DEL”) is seen in about 20% of DLBCL. These patients
have a worse clinical outcome compared to DLBCL negative for both MYC and
BCL2 or positive for only one of these markers. Recent data suggest that the poor
prognostic significance of MYC-rearranged or MYC-high DLBCL is associated with
inferior outcome only when BCL2 is also rearranged or co-expressed. In addition,
the negative prognostic significance of activated B-cell (ABC) subtype vs. germinal
center B-cell (GCB) subtype in DLBCL appears to be largely explained by MYC/BCL2
| 17-22 October 2014
| EAHP - 2014
|
55
th
double-expression, since this co-expression occurs more commonly in the ABC subtype. Currently, these immunohistochemical (IHC) markers are increasingly used as
part of risk stratification of DLBCL but do not directly impact therapy (Johnson NA et
al., J Clin Oncol 2012; Hu S et al., Blood 2013).
In this follow-up study, we analyzed the clinicopathologic features associated with
activated BCR signaling as determined by qIF in a large cohort of primary DLBCL
samples (N=93), in order to further characterize the significance of activated BCR
signaling in DLBCL and to determine whether there is a link between active BCR
signaling and MYC and BCL2 expression by immunohistochemistry. Of 144 clinical
specimens examined, 93 fulfilled criteria for measurement of pLYN, <pSYK,pBTK>
and Fcyt. IHC stains for BCL2 and MYC were scored independently by two hematopathologists in 10% increments. DEL was defined as tumors demonstrating >40%
MYC and >50% BCL2 expression.
The 93 patients included 57 males and 36 females with a mean age of 62 years.
High MYC expression by immunohistochemistry (>40%) was seen in 27 cases
(29%). Age and gender did not differ significantly between the MYC-high and MYClow subgroups. MYC expression was positively correlated with BCL2 expression as
well as various markers of activated BCR signaling, including an increase in mean
pLYN, <pSYK,pBTK> and Fcyt scores. In addition, DEL cases showed greater BCR
activation as compared to DLBCL lacking expression of both MYC and BCL2. These
findings suggest that the BCR signaling pathway may be more active in MYC-high
DLBCL and DEL compared to other DLBCL subgroups and should be investigated as
a rational therapeutic target in these aggressive subgroups of DLBCL.
Interestingly, all 6 Hi positive patients (7 samples) showed polyclonal immunoglobulin gene rearrangements by PCR, nevertheless 5 cases harboured monotypic B cells
by FCM and / or immunohistochemistry (4 of 5 lambda, only 1 of 5 kappa, see Table).
In contrast, all 3 Hi negative cases were monoclonal by PCR.
Based on the work of the group of Riesbeck (Lund, Sweden) who demonstrated
interaction between Hi and IgD, we suggest that these marginal zone lesions are
caused by Hi infection (or similar bacteria) with stimulation through interaction
with IgD and TLR9. This may result in a marked marginal zone hyperplasia, and in
some patients ultimately in a monoclonal B cell population with loss of detectable
organisms.
Keywords: Lymphoma, hyperplasia, paediatric
Table 1. Haemophilus influenzae versus B cell clonality in paediatric
marginal zone lesions
Haemophilus Influenzae*
IG PCR**
Keywords: DLBCL, B-cell receptor signaling, double expressing lymphomas
FCM & IHC ***
[OP-LYMP-04]
PAEDIATRIC MARGINAL ZONE LYMPHOPROLIFERATIVE
DISORDER OF THE NECK: A HAEMOPHILUS INFLUENZAE
DRIVEN IMMUNE DISORDER?
Philip M. Kluin1, Ton Langerak2, Janetta Beverdam3, Lydia Visser1,
Joop Van Baarlen4, King Lam5, Kees Seldenrijk6, Robby Kibbelaar7,
Peter De Wit8, Ed Schuuring1, Stefano Rosati1, Arjan Diepstra1,
Max M. Van Noessel9, Jacco C. Hunting10, Mels Hoogendoorn11,
Ellen Van Der Gaag12, Eveline De Bont13, Hanneke C. Kluin14,
Jerome Lo Ten Foe15, Adri Van Der Zanden3
1
Department of Pathology and Medical Biology, University Medical Center Groningen, University of
Groningen, The Netherlands
2
Department Immunology, Erasmus Medical Center Rotterdam, EMCR, Rotterdam, The Netherlands
3
Labmicta, section of Molecular Microbiology, Hengelo, The Netherlands
4
Department Pathology, LPON, Hengelo, The Netherlands
5
Department Pathology, Erasmus Medical Centre Rotterdam, EMCR, Rotterdam, The Netherlands
6
Department Pathology, St Antonius Hospital, Nieuwegein, The Netherlands
7
Department Pathology, Pathology Friesland, Leeuwarden, The Netherlands
8
Department Pathology, Amphia Hospital, Breda, The Netherlands
9
Department Oncology & Hematology, Sophia Children Hospital, Rotterdam, Netherlands
10
Department Internal Medicine, St Antonius Ziekenhuis, Nieuwegein, The Netherlands
11
Department Internal Medicine, Medisch Centrum Leeuwarden, The Netherlands
12
Department Paediatrics, Zorggroep Twente, Hengelo, The Netherlands
13
Department Paediatric Oncology & Hematology, University Medical Center Groningen, University of
Groningen, Netherlands
14
Department Hematology, University Medical Center Groningen, University of Groningen, Netherlands
15
Department Medical Microbiology, University Medical Center Groningen, University of Groningen,
Netherlands
Paediatric nodal marginal zone lymphoma is a rare entity, often characterized by
isolated cervical lymphadenopathy and an indolent course of disease. We describe
12 patients, median age 12.5 yr, with cervical lymphadenopathy (1 bilateral) and a
spectrum of atypical marginal zone hyperplasia / lymphoma. Treatment consisted of
watchful waiting (N=9), local radiotherapy (N=2) and steroid therapy (N=1). With a
median follow up of 67 months, no progression to lymphoma was observed in any of
the 12 patients. Structures mimicking progressively transformed germinal centres
with expansion of IgMD+ marginal zone B cells were frequent.
Eleven cases contained monotypic or monoclonal B cells by flowcytometry (5/5),
immunohistochemistry (7/12), complete immunoglobulin (IG) gene PCR analysis for
IGH, IGK and IGL (5/12) or karyotyping (2/4), with a remarkable inconsistence between immunological techniques and IG gene PCR.
In 4 lymph nodes of 3 patients microbiological analysis was positive for haemophilus influenzae (Hi), in one classified as non typeable (NTHi) biotype III. Quantitative
PCR for Hi with primers for the Hi gyrase gene on frozen material showed positive
results in 5/8 cases tested. In total 6/9 patients (7/10) samples were Hi positive.
56
The other 3 cases could not be tested. Comparison with dilution experiments of a
positive control suggested the presence of a few thousand bacteria per frozen tissue section in the positive cases. Twenty cervical lymph nodes from age matched
controls with non-specific lymphadenitis and 6 other control samples were negative
by Hi PCR. In these controls input of sufficient cellular DNA was controlled by quantitative PCR for human beta globin.
| EAHP - 2014
| 17-22 October 2014
Monoclonal
Weak
Polyclonal
Kappa
Lambda
Polytypic
Positive N=6
0
1
5
1
4
1
Negative N=3
3
0
0
1
0
2
Not done N=3
2
0
1
1
1
1
* combination of culture and PCR tests; ** in all cases full IG BIOMED PCR analysis for IGH, IGK and
IGL; *** in all 12 cases immunohistochemistry, in 5 cases also flowcytometry was performed
[OP-LYMP-05]
KLF2 MUTATION IS THE MOST FREQUENT SOMATIC CHANGE
IN SPLENIC MARGINAL ZONE LYMPHOMA AND IDENTIFIES A
SUBSET WITH DISTINCT GENOTYPE
Alexandra Clipson1, Ming Wang1, Laurence de Leval2,
Margaret Ashton-Key3, Andrew Wotherspoon4, George Vassiliou5,
Niccolo Bolli5, Sarah Moody1, Leire Escudero Ibarz1,
Güneș Gündem6, Kim Brugger7, Anthony Bench8, Mike Scott8,
Hongxiang Liu9, George Follows8, Eloy F. Robles10,
Jose Angel Martinez Climent10, David Oscier11, A. James Watkins12,
Ming Qing Du1
1
Division of Molecular Histopathology, Department of Pathology, University of Cambridge, UK
Institute of Pathology, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland
Department of Cellular Pathology, Southampton University Hospitals National Health Service Trust,
Southampton, UK
4
Department of Histopathology, Royal Marsden Hospital, London, UK
5
Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK;
Department of Haematology, Addenbrooke’s Hospital, Cambridge University Hospitals NHS
6
Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK
7
8
9
Molecular Malignancy Laboratory, Addenbrooke’s Hospital, Cambridge University Hospitals NHS
10
Division of Oncology, Center for Applied Medical Research CIMA, University of Navarra, Pamplona,
Spain
11
Department of Haematology, Royal Bournemouth Hospital, Bournemouth, UK
12
Division of Molecular Histopathology, Department of Pathology, University of Cambridge, UK;
2
3
To characterise the genetics of splenic marginal zone lymphoma (SMZL), we performed whole exome sequencing of 16 cases and identified novel recurrent inactivating mutations in KLF2, a gene whose deficiency was previously shown to cause
splenic marginal zone hyperplasia in mice. KLF2 mutation was found in 40 (42%)
of 96 SMZLs, but rarely in other B-cell lymphomas. The majority of KLF2 mutations
were frameshift indels or nonsense changes, with missense mutations clustered in
In conclusion, this study shows that atypical CD10-negative and/or BCL2-negative
FL are cytogenetically heterogeneous, comprising a subset harbouring del 1p36
(28%) as the sole genetic alteration or in association with BCL2 or BCL6 or IGH
breaks. Further studies are needed to better characterize the remaining subsets of
so called FL lacking any genetic alteration detectable with the present approach. We
also validate in the present study the use of an automatic counting for the detection
of del 1p36 on FFPE tissue sections that will facilitate the detection of this abnormality in routine practice
Keywords: atypical follicular lymphoma, 1p36 deletion, heterogeneous
Keywords: SMZL, KLF2 mutation, IGHV1-2
[OP-LYMP-07]
[OP-LYMP-06]
ATYPICAL CD10-NEGATIVE AND/OR BCL2-NEGATIVE
FOLLICULAR LYMPHOMA ARE GENETICALLY HETEROGENEOUS
AND COMPRISE A SUBSET WITH 1P36 DELETION
BLASTIC PLASMACYTOID DENDRITIC CELL NEOPLASM:
MOLECULAR HIGH-THROUGH-PUT TECHNOLOGIES SHED
NEW LIGHT ON ITS HISTOGENESIS, PATHOBIOLOGY AND
THERAPEUTIC OPTIONS
Vanessa Szablewski1, Maryse Baia2, Christian Bastard3,
Christine Terre4, Teresa Marafioti5, Jean Michel Picquenot6,
Claire Glaser7, Marie Hélène Delfau Larue8, Jehan Dupuis9,
Corinne Haioun9, Philippe Gaulard10, Christiane Copie Bergman10
Stefano Aldo Pileri1, Valentina Tabanelli1, Federica Melle1,
Antonella Laginestra1, Claudio Agostinelli1, Emilio Berti2,
Lorenzo Cerroni3, Fabio Facchetti4, Fabio Fuligni1, Raul Rabadan5,
Pier Paolo Piccaluga1, Maria Rosaria Sapienza1
1
Département de Biopathologie Cellulaire et Tissulaire des Tumeurs, CHU Montpellier, Hôpital Gui
De Chauliac, Montpellier, France
2
3
INSERM U918, Centre Henri Becquerel, Rouen, France
4
Laboratoire de cytogénétique hémato-oncologique, Centre Hospitalier de Versailles, Le Chesnay,
France
5
Department of Histopathology, University College Hospital London, UK
6
Département de Biopathologie Intégrée du Cancer, Centre Henri Becquerel, Rouen, France
7
Hôpital Mignot, service d’Anatomie Pathologique, Versailles, France
8
9
10
Follicular lymphoma (FL) accounts for 20-30% of adult non-Hodgkin lymphomas.
The histopathological diagnostic criteria are usually straightforward in its classical form. The cytogenetic hallmark is the t(14;18)(IGH-BCL2) translocation observed
in up to 90% of FL, but other genetic alterations have been reported including
BCL6/3q27 rearrangement and 1p36 deletion. FL may also be heterogeneous in
terms of clinical presentation, morphological, immunophenotypical features. These
atypical forms are difficult to diagnose and to differentiate from other low grade
B-cell malignancies. The aim of the study was to better characterize a series of
55 atypical CD10-negative and/or BCL2-negative FL using interphase FISH analysis with BCL2, BCL6, IGH, IGK, IGL breakapart probes, IGH-BCL2 fusion and 1p36
probes. Del 1p36 was evaluated using a dual color probe 1p36/1q25 and both manual and automated scoring on digital slides with the Visilog software (FEI, France)
were performed. The cut-off ratio was established and validated using positive and
negative controls (5 reactive lymphoid tissues, 5 classical t(14;18) FL and 9 del
1p36 FL characterized by CGH microarray and/or conventional cytogenetics). Ten
classical FL and 7 nodal marginal zone lymphomas (MZL) were analysed in parallel
as control.
Patients mean age was 61 years old [30-92] and male:female ratio was 1,2:1. Most
cases were nodal (51/55). FL cases displayed a follicular and/or diffuse pattern of
growth, were composed of centrocytes admixed with a variable number of centroblasts and classified as grade1-2 (49/55) or rarely grade 3A (n=4) or 3B ( n=2), and
had an CD20+, CD5-, CD10- (n=12/55), BCL2- (clone 124 and E17)(47/55), BCL6+
(51/52) immunophenotype. All evaluable cases (54/54) expressed the recently described FL marker STMN1 (stathmin).
Classical t(14;18)(IGH-BCL2) or BCL6 breakpoint were observed in 4/48 (8,3%) and
6/48 (12.5%) of FL. Del 1p36 was detected in 12/43 (28%) of evaluable cases, as
the sole genetic abnormality (n=4), associated with t(14;18)(IGH-BCL2) (n=2), with
BCL6 breakpoint (n=3), or IGH rearrangement (n=3). FL with del 1p36 occurred predominantly in male (54.5%) with a mean age of 59 years with disease in the inguinal nodes (6/12). They were BCL6+ (12/12), CD10+ (8/12), BCL2- (10/12), CD23+
(6/11) and STMN1+ (12/12).We also describe a group of “atypical FL” lacking any
aforementioned genetic alterations detectable with the present approaches accounting for 23/40 (57.9%) of evaluable cases with all probes. In the control group,
all classical FL were associated with BCL2 breakpoint including one case with both
1
Unit of Haematopathology, Bologna University School of Medicine, Bologna, Italy
Dermatology Unit, University of Milan-La Bicocca, Milan, Italy
Department of Dermatology, University of Graz, Graz, Austria
4
Pathology Section, Department of Molecular and Translational Medicine, University of Brescia,
Brescia, Italy
5
Department of Biomedical Informatics and Center for Computational Biology and Bioinformatics,
Columbia University, New York, NY 10032, USA
2
3
BCL2 and BCL6 rearrangement, all nodal MZL were negative with both probes and
del 1p36 was not observed.
the C-terminal zinc finger domains. Functional assays showed that these mutations
inactivated the ability of KLF2 to suppress NF-kB activation by TLR, BCR and TNFR
signalling. Further extensive investigations revealed common and distinct genetic
changes between SMZL with and without KLF2 mutation. IGHV1-2 rearrangement
and 7q deletion were primarily seen in SMZL with KLF2 mutation, while MYD88
and TP53 mutations were nearly exclusively found in those without KLF2 mutation.
NOTCH2, TRAF3, TNFAIP3 and CARD11 mutations were observed in SMZL both with
and without KLF2 mutation. Taken together, KLF2 mutation is the most common genetic change in SMZL and identifies a subset with a distinct genotype characterised
by multi-genetic changes. These different genetic changes may deregulate various
signalling pathways and generate cooperative oncogenic properties, thereby contributing to lymphomagenesis.
Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare disease with dismal prognosis, included among acute myeloid leukaemias in the 2008 WHO
Classification. To date, no standard therapeutic approach has been established
and the refinement of curative strategies for BPDCN definitely requires extensive
molecular studies. Accumulating data about epigenetic control impairment suggest
mutational profile similarity with other myeloid disorders.
To better understand the pathobiology of BPDCN and discover new targets for more
effective therapies, a large series of tumors underwent gene expression profiling
(GEP) (25 samples) and next generation sequencing (NGS) (14 samples).
For the first time, GEP showed that BPDCN originates from resting plasmacytoid
dendritic cells (pDC) of myeloid origin. Notably, its signature revealed two sets of
genes, consisting of 46 components each, which turned out to be deregulated in
common with acute myeloblastic (AML) and lymphoblastic (ALL) leukaemias, respectively. By connectivity map, it was assessed that some of these genes encode
for products targeted by drugs included in ALL protocols, thus explaining the paradoxical efficacy of the latter. Furthermore, an integrated bio-informatic approach
revealed aberrant activation of the NF-kB pathway by suggesting it as a novel therapeutic target. The efficacy of anti-NF-kB-therapy on the BPDCN cell line CAL-1 was
effectively tested, and the molecular shut-off of the NF-kB pathway was successfully demonstrated by GEP and IHC.
WES analysis was performed on four BPDCN samples, identifying recurrent mutations in polycomb complex associated genes (e.g. TET2, ASXL1, and SUZ12). The
observed mutations were validated in 14 tumors by MiSeq targeted re-sequencing
analysis. We hypothesize that these potentially inactivating mutations might lead to
an impaired polycomb repressive complex 2 (PCR2) methyltransferase function. To
this end, we matched the data derived from BPDCN GEP with a list of known PRC2
target genes, identifying 160 genes significantly up regulated in BPDCNs vs. resting
pDCs (p value <= 10-5).
At the time being, further explorative studies are ongoing including the NGS analysis
of 5 additional BPDCNs and the development of a mouse model aiming to test the
efficacy of histone-deacetylase inhibitors, Bortezomib and anti-angiogenetic drugs,
either singly or in combinations.
Supported by a grant AIRC 5x1000 (10007).
Keywords: BPDCN, NGS, epigenetic
| 17-22 October 2014
| EAHP - 2014
|
57
th
[OP-LYMP-08]
EXTRAMEDULLARY PLASMACYTOMAS OF THE UPPER
AERODIGESTIVE TRACT ARE EXTRANODAL MARGINAL ZONE
LYMPHOMAS WITH PLASMACYTIC DIFFERENTIATION
Iris Miedema1, Nathalie Hijmering1, Jan Paul De Boer2,
Olga Balagué Ponz3, Jacqueline Cloos4, Sonja Zweegman4,
Daphne De Jong1
1
VU University Medical Center, dept of Pathology, Amsterdam, the Netherlands
Netherlands Cancer Institute, Dept of Medical Oncology, Amsterdam, the Netherlands
3
Netherlands Cancer Institute, Dept of Pathology, Amsterdam, the Netherlands
4
VU University Medical Center, Dept of Hematology, Amsterdam, the Netherlands
2
Background: There is an ongoing discussion whether extramedullary plasmacytoma (EMP) at MALT-sites in the upper aero-digestive (UED) tract should rather be
considered as extranodal marginal zone lymphoma, MALT-type (MALT-lymphoma)
or rather as related to multiple myeloma (MM). Although in general MALT-lymphoma
and MM have a different clinical presentation and course, morphological and genetic features, discrimination in the UED tract remains challenging. The limited data
that are available are heterogeneous and can support both hypotheses.
Figure 1. Three morphological groups: 3 study groups, covering the
morphological spectrum of MALT-lymphoma/MM in the upper aerodigestive tract
Methods: From the files of the Comprehensive cancer Center Amsterdam, 41 patients diagnosed and treated (1993-2013) under the dagnosis of MALT-lymphoma,
lymphoplasmacytic lymphoma or EMP with primary presentation in the oral cavity,
pharynx, larynx, nasopharynx and nasal cavity of which biopsy material and complete clinical information was available, were included. Immunohistochemical stainings were performed for CD20, CD79a, CD138, CyclinD1, BCL2, CD10, BCL6, CD23,
IgH classes and pankeratin to assess the presence of lymphoepithelial lesions (LEL).
All biopsy samples at presentation and follow-up were reviewed. Genetic alterations
were monitored using RT-PCR for API2-MALT1 translocation, PCR /sequencing for
MYD88 L265P mutation. In selected cases, a CCND1 BA_FISH assay was performed
to confirm translocation. Further FISH analysis is currently performed (IgH, MALT1,
MM FISH).
Results: Histological review showed a spectrum of cellular composition with 7
cases with a predominant small B-cell infiltrate and marginal zone differentiation
(group 1), 21 cases with pure plasmacytic morphology (EMP, group 3) and 13 cases
with a mixed composition with varying components of lymphocytic, plasmacytoid
and plasmacytic morphology (group 2). LEL were observed 9/16 group 1 and 2
cases containing glandular epithelium, but also in 3/12 cases with EMP containing
microscopic clusters of CD20+ lymphocytes. In none of the cases, hoewever, the
plasmacells invaded the epithelium to form LEL. Genetic screening showed API2MALT1 translocation in only one patient (group 1), whereas no MYD88 mutations
were found. One case with MALT-lymphoma morphology showed heterogeneous
CCND1 overexpression with a subclonal CCND1 break by FISH.
Clinically, patients presented with clinical features of MALT-lymphoma with autoimmune disease (n=5, Sjogren, TTP, RA), localisations at other MALT-sites (n=2) or
regional lymphadenopathy at presentation or follow-up (n=7) (follow-up 1-20 years,
mean 5.2 years) with transformation to DLBCL in 2. These features were equally
presented in all morphological groups. No patient presented with lytic bone lesions
or developed features of MM.
Conclusion: This study provides histological and clinical evidence that so-called
EMP of the upper aero-digestive tract should rather be considered as MALT- lymphoma, while the negative genetic evidence does not argue against this notion (further FISH analyses pending).
Localised EMP is generally treated with high dose radiotherapy (40-50Gy). Since
the aim is curative intent, significant morbidity with scarring with functional (vocal) impairment is accepted. The preferred first line treatment for localised MALTlymphoma is either indolent-type chemotherapy or low-dose radiotherapy, which
results in significantly less early and late morbidity and with the present insights
this treated should be seriously considered in these patients.
Keywords: extramedullary plasmacytoma, MALT-lymphoma, upper-aero-digestive
tract
Figure 2. Plasmacytoma upper aerodigestive tract. Lymphoepithelial lesion in a
laryngeal plasmacell proliferation.
58
| EAHP - 2014
| 17-22 October 2014
Ming Wang1, Leire Escudero Ibarz1, Sarah Moody1, Naiyan Zeng1,
Alexandra Clipson1, Yuanxue Huang1, Xuemin Xue1,
Nicholas F. Grigoropoulos1, Sharon Barrans2, Lisa Worrillow2,
Tim Forshew3, Francesco Marass3, Nitzan Rosenfeld3, Jing Su3,
Andrew Firth1, Howard Martin4, Andrew Jack2, Kim Brugger4,
Ming Qing Du1
1
Department of Pathology, University of Cambridge, Cambridge, UK
Haematological Malignancy Diagnostic Service, St. James’s Institute of Oncology, Leeds, UK
3
Cancer Research UK Cambridge Institute, Li Ka Shing Centre, University of Cambridge, Robinson
Way, Cambridge, UK
4
2
The advent of next generation sequencing (NGS) has lead to unprecedented discoveries in somatic mutation profiling of human cancer. One of the major challenges
is to validate a large number of somatic mutations using archival formalin-fixed
paraffin-embedded (FFPE) tissue biopsies by targeted re-sequencing due to a lack
of established experimental protocols and methodically validated variant calling algorithms. To address these issues, we developed a high throughput mutation screen
by multiplex PCR with Fluidigm Access Array and Illumina MiSeq sequencing, and
established a companion variant calling algorithm. The experimental protocol and
variant calling algorithm were developed and validated against the somatic mutations in 7 genes detected by Sanger sequencing of DNA samples from a total of 163
FFPE diffuse large B-cell lymphoma biopsies. In the initial study, 111 PCR primer
pairs for the 7 genes covering 21kb sequence were designed, and their combination
for pre-amplification of template DNA and subsequent multiplex PCR with Fluidigm
Access Array were guided by in silico PCR and AutoDimer software analyses, and
further verified by experiments. A series of quality control measures were established at various steps of Fluidigm PCR/MiSeq sequencing including quality control
of template DNA, pre-amplification, Fluidigm PCR, barcode labelling and library purification. An in-house variant calling algorithm was developed and optimised against
a total of 151 known somatic mutations detected by Sanger sequencing, including 138 substitutions and 13 indels (ranging 1-33bp). In the subsequent study, the
above optimised experimental protocol and variant calling algorithm were tested in
two separate experiments. In one experiment, the above 111 PCR primer pairs for
the 7 genes were investigated as a part of a total of 344 PCR primer pairs for 22
genes covering 65kb sequence using the above cohort of FFPE DNA samples. The
mutations detected in this independent experiment were highly concordant with
those seen in the initial validation study. In the other experiment, the above 111 PCR
primers for 7 genes were analysed as a part of 157 PCR primer pairs for 13 genes
in an additional cohort of 38 high molecular weight DNA samples from lymphoma,
and this experiment was performed twice independently. The two independent experiments showed a complete concordance in mutation detection. In conclusion,
we have developed and validated a robust high throughput mutation screen using
DNA samples from archival FFPE tissues by Fluidigm multiplex PCR/Illumina MiSeq
sequencing, and an in-house variant calling algorithm.
We investigated the protein expression of GPER and the phosphorylation status of
downstream targets in primary cases and cell lines. GPER was expressed in 49 of
87 (56%) primary MCL cases and in 10 of 13 (77%) MCL cell lines, whereas only 6
of 20 (30%) of diffuse large B-cell lymphomas and only 1 of 20 (5%) follicular lymphomas were positive for GPER. In MCL, immunohistochemical expression of GPER
did neither correlate with proliferation, assessed by Ki-67 staining, nor with subtype.
In cell lines with high level of GPER, however, inhibition of GPER or siRNA induced
downregulation of GPER abrogated or reduced the phosphorylation of p42/44 MAPK
and AKT. Interestingly, the expression of CyclinD1 was reduced as well, which led
to cell cycle arrest and inhibition of cell proliferation. Moreover inhibition of GPER
induced apoptosis as shown by cleavage of caspase3. In cell lines with low level of
expression of GPER the inhibition of GPER had no effect. Given that GPER can increase microtubulus dynamics, we wanted to test if resistance to Taxol (Paclitaxel),
which strongly suppresses microtubulus dynamics, can be overcome by inhibition
of GPER. Indeed, inhibition of GPER and treatment with Paclitaxel was highly synergistic for inducing apoptosis in MCL cell lines.
These data suggest that GPER may be involved in the pathogenesis of MCL and inhibition of GPER may be effective in the treatment of MCL, especially in combination
with microtubulus stabilizing agents like Taxol.
Keywords: mantle cell lymphoma, signaling pathway, G protein-coupled receptor
SOMATIC MUTATION SCREENING USING ARCHIVAL FORMALINFIXED PARAFFIN-EMBEDDED TISSUES BY FLUIDIGM ACCESS
ARRAY MULTIPLEX PCR AND ILLUMINA MISEQ SEQUENCING
individual genes involved in signaling pathways, that affect cellular proliferation
have been identified as overexpressed or underexpressed in MCL. G protein-coupled receptors, like GPER, can stimulate diverse signaling pathways leading to cell
migration, survival, microtubulus destabilization and proliferation in normal cells
and various cancer cell types. Therefore we wanted to determine whether GPER is
involved in the pathogenesis of MCL
[OP-LYMP-09]
Figure 1. GPER is expressed in the majority of primary MCL cases (A) and MCL
cell lines (B)
Keywords: FFPE DNA, Fluidigm PCR, MiSeq sequencing
[OP-LYMP-10]
THE G PROTEIN-COUPLED ESTROGEN RECEPTOR 1 (GPER)
CONTRIBUTES TO THE PROLIFERATION AND SURVIVAL OF
MANTLE CELL LYMPHOMA
Figure 2. Inhibition (A) or siRNA mediated downregulation (B) of GPER results in abrogation of phosphorylation of p42/44 and AKT as well as in induction
of apoptosis in MINO, but has no effect in MAVER (A).
Martina Rudelius1, Elena Hartmann1, Hilka Rauert Wunderlich1,
Wolfram Klapper2, German Ott3, Andreas Rosenwald1
1
Institute of Pathology, Julius-Maximillians-University Wuerzburg, Wuerzburg, Germany
Institute of Pathology, University Hospital Schleswig-Holstein, Campus Kiel, Germany
3
Department of Clinical Pathology, Robert-Bosch-Krankenhaus and Dr. M. Fischer-Bosch Institute of
2
Mantle cell lymphoma (MCL) is an aggressive subtype of B-cell Non-Hodgkin lymphoma, which is generally incurable. It is characterized by the t(11;14)(q13;q32)
translocation involving CCND1, which results in deregulation of cell cycle progression. Survival can be predicted by a set of proliferation-related genes and many
Figure 3. Inhibition of GPER and treatment with Paclitaxel (A) is highly
synergistic in MCL cell lines (B).
| 17-22 October 2014
| EAHP - 2014
|
59
th
[OP-LYMP-11]
and molecular features, little is known about clinico-biologic features of HCV-related
DLBCLs.
FOLLICULAR CYTOTOXIC CD8 T-CELLS (CD8TFC) SHARING
FUNCTIONAL SIMILARITIES WITH CD4TFH CELLS ARE
PRESENT IN CLASSICAL HODGKIN’S LYMPHOMA TISSUES
DISPLAYING A SPECIFIC “MIXED NODULARITY” PATTERN
Herein we investigated a series of HCV-related DLBCLs for NOTCH, NF-B and
B-cell receptor signaling mutations, three signaling pathways involved in normal
MZ development and whose deregulation may play a role in splenic MZL.
Luc Xerri, Suong Le, Sylvaine Just Landi, Françoise Gondois Rey,
Samuel Granjeaud, Daniel Olive
Centre de Recherche en Cancérologie de Marseille, INSERM U1068 and Institut Paoli-Calmettes,
Marseille, France Marseille, France
We have previously reported that some classical Hodgkin’s Lymphoma (cHL) tissues
display a gene signature evocative of a Th1 immune reaction. In order to better
characterize this process, immune cell subsets were isolated from cHL fresh tissue
samples (n=19) using a powerful 11 colors flow cytometry method, in parallel with
cell sorting. Fresh tissue samples from follicular B cell lymphoma (FL), diffuse large
cell B cell lymphoma, and reactive lymphadenitis were used as controls (n=23).
In 4 cLH cases, we observed a significant proportion of activated CD8+ T-cells
expressing ICOS and CXCR5 at high levels. The presence of either CD8+/ICOS+/
CXCR5- T cells or CD8+/ICOS +/ CXCR5+ T-cells was a specific feature of cHL tissues since it was absent from B-cell lymphomas and reactive tissues. In contrast,
CD8+/CXCR5+ T-cells were found not only in cHL, but also in most other samples
analyzed.
Further phenotypic characterization showed that the CD8+/ICOS +/ CXCR5+ T cells
expressed markers associated with CD4 TFH cells, like PD1, bcl-6, BTLA, CXCL13
and IL-21. Under stimulation, they expressed only low levels of IFN-G, granzyme B
and perforine. Functional properties of CD8FC cells included the capacity to stimulate B-cell proliferation in a similar degree as CD4 TFH cells. The 4 cHL cases associated with CD8+/ICOS +/ CXCR5+ T-cells contained CD30+ CD15+ EBV+ Reed
Sternberg cells (RSC). They were characterized by a particular “mixed nodularity”
pattern, in which sclerosis was absent. Various nodules were observed, including
reactive germinal centers (GC) partly colonized by RSC co-localizing with CD8+/
ICOS+ T-cells, suggesting an early GC invasion triggering an intra-follicular CD8
T-cell reaction. Other nodules were composed of a high number of RS cells admixed
with numerous CD8+/ICOS+ T-cells. This “mixed nodularity” pattern was absent in
the other HL cases.
Altogether, our results point out a previously unrecognized intra-follicular CD8 T-cell
subset sharing phenotypic and functional features with CD4 TFH, that we have
thus considered as putative “follicular cytotoxic” CD8 T-cells (TFC). This cell subset
appears to be specifically associated with EBV+ cHL tissues with unusual histophenotypic features, which may probably reflect a strong CD8 activation process.
Results: Histological examination of HCV-related DLBCLs revealed centroblastic
morphology in most cases (37/46, 80.4%). In 12/46 (26%) cases a minor area of
the diagnostic biopsy was composed of small to medium sized monocytoid B-cells,
suggesting a progression from an underlying MZL.
In HCV-positive cases, NOTCH2 mutations were detected in 9/46 (20%) and NOTCH1
mutations in 2/46 (4%), respectively. In the control group, NOTCH2 was mutated
respectively in 1/64 (2%), whereas search for NOTCH1 mutation was negative in
all cases. The comparison between the two groups was statistically significant (p
= 0.002) for NOTCH2. NOTCH pathway mutations were enriched in cases harbouring a small to medium cells component, histologically resembling MZL, (6/12, 50%
in cases with low grade component vs 6/34, 17.7% in cases without low grade
component; p=0.05). In our panel, the prevalence and type of NF-kB pathway mutations was similar with the ones of DLBCL of non-GC phenotype arising in the HCVnegative subject. In the other hand, BCR pathway (CARD11, CD79A and CD79B) was
never affected by genetic lesions.
Moreover, 5-years OS was worse in pts with NOTCH1 or NOTCH2 mutations (29%;
95% CI: 5%-59%) than in pts without them (60%; 95%CI: 41%-75%). In multivariate analysis, NOTCH pathway disruption retained its prognostic effect also adjusting
for IPI score and histogenesis (HR=7.9; 95%CI: 1.2-51; p=0.029). When adjusting
also for age (as continuous variable), NOTCH pathway was still significant (HR=9.0,
95% CI: 1.3-60.1; p=0.024).
Conclusion: Our study showed that, in HCV-positive DLBCLs, a significantly high
rate of cases harbours mutations of NOTCH genes and that the disruption of this
pathway is associated with a worse OS. Both these findings corroborate the hypothesis that HCV-positive DLBCLs carrying NOTCH mutations may actually represent
the transformed phase of a MZL clone.
Keywords: Hodgkin’s lymphoma, CD8, ICOS
Keywords: Diffuse large B cell lymphoma; HCV infection; NOTCH pathway mutation;
marginal zone lymphoma
[OP-LYMP-12]
[OP-LYMP-13]
NOTCH PATHWAY DISRUPTION IN A SUBSET OF HCV-RELATED
DIFFUSE LARGE B CELL LYMPHOMA: ITS ASSOCIATIONS TO
PROGNOSIS AND TO A POSSIBLE MARGINAL ZONE DERIVATION
UNRAVELING THE ROLE OF CD4 T CELLS IN HODGKIN
LYMPHOMA
1
2
3
1
Marco Lucioni , Luca Arcaini , Davide Rossi , Marta Nicola ,
Roberta Riboni1, Antonio Ramponi4, Valeria Virginia Ferretti2,
Maurizio Bonfichi2, Manuel Gotti2, Aldo Maffi1, Giorgio Alberto Croci1,
Mariarosa Arra1, Valeria Fiacacdori2, Marzia Varettoni2,
Sara Rattotti2, Lucia Morello2, Elena Dallera1, Gianluca Gaidano3,
Mario Cazzola2, Marco Paulli1
1
Anatomic Pathology Unit, Department of Molecular Medicine, University of Pavia, Pavia, and
Pathology Unit, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy Scientifico (IRCCS) Policlinico
San Matteo, Pavia, Italy
2
Division of Hematology, Department of Molecular Medicine, University of Pavia and Fondazione
Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS) Policlinico San Matteo, Pavia, Italy
3
Division of Hematology, Department of Translational Medicine, Amedeo Avogadro University of
Eastern Piedmont, Novara, Italy
4
Division of Pathology, Department of Health Science, Amedeo Avogadro University of Eastern
Piedmont, Novara, Italy
Background: It is widely accepted that Hepatitis C virus (HCV) is significantly associated with B-cell non-Hodgkin’s lymphomas (B-NHL), mostly marginal zone B-cell
lymphomas (MZL) and diffuse large B cell lymphoma (DLBCL). While HCV-related
MZLs have been extensively characterized displaying distinctive clinicopathologic
60
Material and methods: On the basis of the availability of adequate frozen and/or
paraffin embedded specimens, we tested for NOTCH, NF-B and B-cell receptor
signaling mutations 46 cases of HCV-related DLBCLs and 64 HCV-negative DLBCLs,
for comparison. The mutation hotspots of NOTCH1, NOTCH2, SPEN, MYD88, BIRC3,
IKBKB, TNFAIP3, TRAF3, CARD11, CD79A and CD79B were analyzed by PCR amplification and DNA direct sequencing of genomic DNA. Purified amplicons were subjected to conventional DNA Sanger sequencing using the ABI PRISM 3100 Genetic
Analyzer (Applied Biosystems), and compared to the corresponding germline sequences using the Mutation Surveyor Version 4.0.5 software package (SoftGenetics)
after automated and/or manual curation.
| EAHP - 2014
| 17-22 October 2014
Benjamin Rengstl1, Frederike Schmid1, Christian Weiser1,
Claudia Döring1, Tim Heinrich1, Kathrin Warner2, Petra S.A. Becker3,
Christian Seidl3, Hinrich Abken2, Ralf Küppers4, Martin Leo Hansmann1,
Sebastian Newrzela1
1
Dr. Senckenberg Institute of Pathology, Medical School, Goethe-University of Frankfurt, 60590
Frankfurt am Main, Germany
2
Department I of Internal Medicine, Medical School, University of Cologne, 50937 Cologne,
Germany
3
Institute for Transfusion Medicine and Immunohematology, Red Cross Blood Donor Service,
Baden-Württemberg-Hessen, 60528 Frankfurt am Main, Germany
4
Institute of Cell Biology (Cancer Research), Medical School, University of Duisburg-Essen, 45122
Essen, Germany
Hodgkin lymphoma (HL) presents with a unique histological pattern. Within affected tissue the pathognomonic Hodgkin and Reed-Sternberg (HRS) cells account for
only less than 1 % of the tumor mass and are embedded in a reactive infiltrate
mainly consisting of CD4 T cells. To escape anti-tumor immunity, HRS cells have
to modulate the surrounding cells inducing an immunosuppressive microenvironment. We performed co-culture studies of HRS cells and primary human immune cells identifying a strong anti-tumor potential of T cells in vitro and in vivo.
Surprisingly, anti-HRS cell reactions were mediated by CD4 and not cytotoxic T
cells. We further investigated the molecular make-up between HRS and CD4 T cell
Table 1. BCL6 immunohistochemistry and FISH results
Keywords: Hodgkin lymphoma, immunotherapy, chimeric antigen receptors
Table 2. Other immunohistochemical markers in cases with BCL6
chromosomal breaks
[OP-LYMP-14]
BCL6 PROTEIN EXPRESSION AND BCL6 CHROMOSOMAL
BREAKS IN NODAL MARGINAL ZONE LYMPHOMA WITH
DIAGNOSTIC IMPLICATIONS
Michiel Van Den Brand, Patricia Groenen, Konnie Hebeda,
Han Van Krieken
Radboud University Medical Center, Nijmegen, The Netherlands
Nodal marginal zone lymphoma (NMZL) is a rare type of B-cell lymphoma with a
largely unknown pathogenesis. It can be difficult to distinguish morphologically and
immunohistochemically from follicular lymphoma (FL). BCL6 protein expression as
shown by immunohistochemistry is used in diagnostic practice to support a diagnosis of FL. So far, studies in NMZL frequently used expression of germinal center
markers (including BCL6) as an exclusion criterion, thereby potentially introducing
bias.
Table 3. Other immunohistochemical markers in cases with BCL6 protein
expression
interactions. Interestingly, we were able to detect an adhesion complex between
both cell types that showed a high similarity to immunological synapses formed
between antigen presenting cells and T cells. In line with this, we could show
that the anti-tumor reaction is based on the recognition of major histocompatibility complex class II (MHC-II) presented by HRS cells. Using primary cells from
MHC-II compatible donors of HRS cells, we were able to abrogate the anti-tumor
potential of CD4 T cells. However, gene expression profiling of co-cultured HRS
cells as well as in vivo tumor infiltration of CD4 T cells revealed that both cell
types continuously interact with each other even in the MHC-II compatible setting.
Next, we introduced an HRS-cell specific chimeric antigen receptor into MHC-II
compatible CD4 T cells restoring their anti-tumor potential and leading to potent
HRS-cell killing. Our work gives insight into the interactions between HRS cells
and their microenvironment indicating CD4 T cells to be perfectly suited for an
immunotherapeutic approach in HL.
In this study, we tested 47 cases that were primarily diagnosed as NMZL for BCL6
protein expression and the presence of chromosomal breaks involving BCL6. All
cases lacked chromosomal breaks involving BCL2. BCL6 protein expression, as
shown by immunohistochemistry, was detected in 8 out of 47 cases (17%, Table 1).
Fluorescent in situ hybridization showed chromosomal breaks involving BCL6 in 8
out of 47 cases (17%, Table 1). However, BCL6 protein expression and the presence
of a BCL6 break showed very poor correlation with only 1 case showing both a BCL6
break and BCL6 protein expression.
The eight cases with a chromosomal break involving BCL6 showed different patterns with additional immunohistochemical markers; two showed expression of germinal center markers (BCL6, CD10, LMO2 and/or HGAL), two showed expression of
both germinal center and marginal zone markers (MNDA and/or IRTA1), two showed
expression of marginal zone markers and two cases were negative for all additional
markers tested (Table 2).
Of the eight cases with BCL6 protein expression, four showed expression of additional germinal center markers (CD10, LMO2, and/or HGAL) but not marginal zone
markers (MNDA and/or IRTA1)(Table 3). The four other cases showed positivity for
marginal zone markers, providing another argument that these four cases represent
actual NMZLs.
These results suggest that the group containing cases with BCL6 breaks and/ or
protein expression is heterogeneous and contains cases of NMZL, but there may
also be BCL2 break negative cases in the gray area between NMZL and FL.
In conclusion, we show that both BCL6 expression and the presence of chromosomal breaks involving BCL6 occur in a subset of lesions that were diagnosed as NMZL.
The fact that NMZLs occasionally express BCL6 and/or CD10 has implications for
hematopathological practice: instead of relying on single markers to differentiate
between FL and NMZL, more criteria should be used to make this distinction.
Keywords: Nodal marginal zone lymphoma, BCL6, Follicular lymphoma
[OP-LYMP-15]
THE PRESSURE OF THE ANTIGENS IN THE PATHOGENESIS OF BL.
EVIDENCE FROM NGS ANALYSIS OF BCR
Cristiana Bellan1, Teresa Amato1, Pier Paolo Piccaluga2,
Francesco Abate2, Giulia De Falco1, Maria Raffaella Ambrosio1,
Raul Rabadan3, Stefano Pileri2, Lorenzo Leoncini1
1
Anatomical Pathology Section, Department of Medical Biotechnology, University of Siena, vie delle
Scotte, 6, 53100, Siena, Italy
2
Molecular Pathology Laboratory, Department of Experimental, Diagnostic, and Specialty Medicine,
Bologna University Medical School, Unit of Hematopathology, S. Orsola Malpighi Hospital, Via
Massarenti 9, 40138 Bologna, Italy.
3
Department of Biomedical Informatics, College of Physicians and Surgeons, Columbia University,
New York, USA
B cell receptor (BCR) is essential for normal B cell development and maturation.
There is increasing evidence that BCR signaling is implicated in B cell malignancies.
Among the different types of B cell malignancies, several different tactics are applied for activation of the BCR pathway.
Roles for antigen-dependent and antigen independent BCR signaling have now been
described for several different lymphoma types.
Intriguingly, recent studies using high-throughput RNA sequencing and RNA interference screening disclosed the importance of BCR pathway in the pathogenesis of
BL. In fact mutations affecting the transcription factor TCF3 or its negative regulator
ID3 were reported in about 70% of sporadic BL cases. This results in the activation
of a tonic form BCR signaling and increased expression of genes belonging to BCR
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However the tonic form of BCR is apparently antigen independent and it is in contrast
with the epidemiological contest of chronic antigen stimulation in which eBL arise.
Table 1. Results of sequence data analysis
Since no definitive data are available at the moment on the mutations affecting
the transcription factor TCF3 or its negative regulator ID3 for eBL, in the present
study we have first assessed the frequency of TCF3 and ID3 in eBL and compared
with sBL. ID3 SNVs were found in 6/26 eBL cases (23%) occuring more frequently
than TCF3 SNVs (3/26; 11%). One sample had mutations in both genes. This rate of
mutations is significantly lower than that reported in sBL 70%.
We then analysed the BCR status by NGS to answer the question whether an activation of B cell receptor through an extrinsic signaling antigen driven could be
demonstrated in BL.
NGS analysis of BCR revealed extensive intraclonal diversification through an active targeted SHM process in eBL. In fact, NGS analysis revealed the presence of
multiple clones sharing the same VDJ and CDR3 in eBL as compared to sBL that
carryed shared as well as unique somatic mutations. The analysis of rearranged VH
genes showed that all cases of eBL constantly harbor mutated VH genes (average
mutation frequency varied from 79.81 % to 97.76 % (median 90,54%); significantly
different from sBL in which mutational variation compared to the germline ranged
from 90.88% to 99.54% ( median 96.6%).
In addition, most of the VH clones possessed any mutations, deletion or insertion
that would prevent or limit translation. It is tempting to speculate that the maintenance of immunoglobulin expression and immunoglobulin –mediated clonal selection have a similar bias. It is noteworthy that our IgVH gene usage analysis showed
that eBL cases have a preferential usage of the VH3-23, VH1-69, and VH4-34 genes.
All together, these findings suggest a strong antigen pressure, that could potentially
be related to a prolonged microenviroment interactions, by whom chronic stimulation of BCR with certain pathogens could be important in promoting clonal expansion of B cells expressing distinctive BCRs and growth of the neoplastic clone.
However we cannot exclude a combining tonic and extrinsic BCR signaling activation in a subset of eBL.
In conclusion, our data support the view that eBL may be considered an infectious/
polymicrobial disease in which the pressure of the antigens play a role and may
arise from pathogenetic pathways that are partially distinct from those driving sBL.
Decifering the pathway of BCR activation in BL may be useful to unravel to which
kind of targeted therapies BL would respond.
Keywords: BCR, Burkitt Lymphoma, NGS
[OP-LYMP-16]
GENOME-WIDE ANALYSIS OF ENHANCER ACETYLATION IN
DLBCL AND MANTLE CELL LYMPHOMA
Russell J.H. Ryan1, Cem Sievers1, Jasleen Kaur1, Holly Whitton2,
Robbyn Issner2, Charles Epstein2, Bradley E. Bernstein1
1
Department of Pathology, Massachusetts General Hospital, Boston, Massachusetts, United States
Epigenomics Initiative, Broad Institute of Harvard University and M.I.T, Cambridge, Massachusetts,
United States
2
Background: Enhancers are distal regulatory elements that play a critical role in
the regulation and maintenance of tissue- and stage- specific gene expression. B
cell lymphomas frequently demonstrate mutation of genes encoding enhancer-regulating proteins (histone-modifying enzymes and sequence-specific transcription
factors (TFs)), suggesting that enhancer dysregulation plays a role in lymphomagenesis. Several classes of emerging therapeutic agents may act through the targeting
of enhancers, including inhibitors of histone deacetylases, histone acetyltransferses,
and acetyl-binding proteins (BET inhibitors).
Approach: We have used chromatin immunoprecipitation sequencing (ChIP-Seq) to profile enhancer-associated histone acetylation (H3K27ac) in three purified normal human B
cell populations and 16 human B lymphoid cell lines, including lines derived from mantle
cell lymphoma (MCL, n=4) and diffuse large B cell lymphoma (DLBCL, n=10).
Figure 1. NGS showed the presence of multiple clones sharing the same VDJ
and CDR3 in eBL as compared to sBL. a, A: capillary electrophoresis analysis; b, B:
NGS results; c, C: sequences of clusters.
Results: Genome-wide acetylation data was analysed by two complementary approaches: the first focusing on individual enhancer units (nucleosome-depleted region (NDR) analysis) and the second focusing on large regions of highly acetylated
chromatin (“superenhancer” analysis). Both approaches were effective in clustering
of populations by phenotype, while regions detected by the NDR approach were
more highly correlated with validated transcription factor binding sites.
Unsupervised clustering revealed a high degree of similarity between enhancer
profiles of primary centroblasts and germinal center phenotype DLBCL lines, while
activated B cell phenotype DLBCL and mantle cell lymphoma lines showed greater
similarity to peripheral blood B cells and tonsil-derived naïve B cells. We identified
numerous enhancers and “superenhancers” specifically associated with the germinal center or non-germinal center state.
Figure 2. A: IgVH gene usage analysis showed that eBL have a preferential
usage of the VH3-23, VH4-34 and VH1-69. B: Evolutionary history of BL 31 case.
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The “superenhancer” region with the most significant differential activity between
mantle cell lines and other populations was located at the CCND1 locus, consistent with aberrant enhancer activation driven by the t(11;14) rearrangement. NDRcentered analysis detected mantle cell lymphoma-specific enhancers adjacent to
SOX11, as well as genes encoding other transcription factors that have not previ-
Keywords: mantle cell lymphoma, chromatin, ChIP-seq
Clinically, all NOTCH mutated cases were diagnosed in female patients, six had
an extra-nodal involvement, and five (71%) had splenic involvement. Significant
differences between wild-type and mutated cases were sex deviation (female proportion: 55% vs. 100%, P=0.04) and splenic involvement (16% vs. 71%, P=0.02).
No significant difference was observed in the overall survival of the patients with
mutated and wild-type FL (5 year survival rate: 71% (NOTCH mutated) vs. 67%
(NOTCH wild-type)).
Discussion: These results indicate that NOTCH mutations are uncommon in FL but
may occur in a subset of cases with particular tumors characterized by low frequency of t(14;18), common splenic involvement and frequent association with DLBCL.
Keywords: Follicular lymphoma, NOTCH, mutation
[OP-LYMP-18]
Figure 1. Hierarchical clustering of populations by H3K27ac enhancer signal.
Hierarchical clustering was performed by identification of H3K27ac+ “superenhancers” for
three primary B cell populations, 16 human lymphoid cell lines, and one myeloid cell line.
[OP-LYMP-17]
RECURRENT MUTATIONS OF NOTCH GENES IN FOLLICULAR
LYMPHOMA IDENTIFY A DISTINCTIVE SUBSET OF TUMORS
Kennosuke Karube1, Daniel Martínez1, Cristina Royo1,
Magda Pinyol1, Paola Castillo1, Alexandra Valera1, Anna Carrió1,
Dolors Costa1, Dolors Colomer1, Maite Cazorla1, Daniel Esteban1,
Andreas Rosenwald2, German Ott3, Eva Giné1,
Armando López Guillermo1, Elias Campo1
1
IDIBAPS, Hospital Clinic, Universitat de Barcelona, Barcdlona, Spain
Institute of Pathology, University of Würzburg, Würzburg, Germany
3
Department of Clinical Pathology, Robert-Bosch-Krankenhaus, and Dr. Margarete Fischer-Bosch
Institute of Clinical Pharmacology, Stuttgart, Germany
2
Background: Follicular lymphoma (FL) is one of the most common malignant lymphomas. The t(14;18)(q32;q21) is found in about 80% of these cases and is regarded to play a principal role in their lymphomagenesis. However, the molecular
mechanisms involved in the development and progression of these tumors are not
fully understood. NOTCH genes play an important role in different steps of T and B
lymphoid differentiation and activating mutations of these genes have been identified in several mature B-cell neoplasms, usually associated with transformation to
large cell lymphoma and aggressive clinical behavior. The role of these mutations in
FL is not known. In this study we have investigated the mutational status of these
genes in FL.
Materials and Methods: A total of 137 samples from 112 FL patients with frozen
material were collected from the pathology files of the Hospital Clínic. Mutational
status of NOTCH1 and NOTCH2 were analyzed using Sanger sequence and allele
specific PCR. These analyses covered the whole PEST domain and most of the TADD
domain, a hotspot region of NOTCH genetic mutations in the B-cell malignancies.
Result: Mutational analysis of NOTCH1 and NOTCH2 was successfully performed
in 112 FL cases. This series includes 71 FL at diagnosis and 34 cases at relapse.
No clinical information was available in 7 cases. NOTCH1 and NOTCH2 truncating
mutations were identified in five and two cases, respectively, (total 7/112, 6.3%). All
truncating mutations predicted for truncated protein in the PEST domain and were
identical to those identified in other B cell lymphoid neoplasms.
In all mutated cases, atypical lymphoid cells had a follicular growth pattern that
was highlighted by the immunohistochemical staining of follicular dendritic cells
DIFFUSE LARGE B-CELL LYMPHOMAS OF IMMUNOBLASTIC
TYPE ARE A MAJOR RESERVOIR FOR MYC-IGH
TRANSLOCATIONS
Annette M. Staiger1, Heike Horn1, Matthias Vöhringer2, Ulrich Hay3,
Elias Campo4, Andreas Rosenwald5, German Ott1, M. Michaela Ott6
Conclusions: Enhancer acetylation profiles of lymphoma cell lines are characterized
by strong signatures of the corresponding developmental state in normal B cells,
while some enhancers show activity specific to certain lymphoma subtypes, suggesting that these profiles may identify lymphoma subtype-specific gene dysregulation. Because enhancer acetylation represents a promising therapeutic target in
lymphoma, a better understanding of enhancer dysregulation in specific lymphoma
subtypes may play a role in targeted therapeutic strategies.
with CD21 and CD23. The comparison of the NOTCH wild-type cases with the seven
NOTCH mutated FL revealed the t(14;18) to occur less frequently in the mutated
cases than in wild-type tumors (14% vs. 69%, P=0.01), whereas no significant differences were identified in immunohistochemical findings (CD10, BCL2 and BCL6).
A DLBCL component associated with the FL at the moment of diagnosis was identified in 4 of the 7 (57%) NOTCH mutated cases but only in 12 of the 64 (18%) wildtype tumors (P=0.03). One mutated FL and 5 out of 64 NOTCH wild-type FL cases
transformed to DLBCL subsequently. These findings suggest that association with
DLBCL is significantly more common in NOTCH mutated than wild-type FL (71%
vs. 23%, P=0.02).
ously been associated with MCL. Other MCL-specific enhancers were located in
association with genes involved in cell trafficking and apoptosis.
1
Department of Clinical Pathology, Robert-Bosch-Krankenhaus’, and ‘Dr. Margarete Fischer-BoschInstitute of Clinical Pharmacology’, Stuttgart, Germany
2
Department of Internal Medicine II, Hematology and Oncology, Robert-Bosch-Krankenhaus,
Stuttgart, Germany
3
Department of Ear, Nose and Throat Surgery, Marienhospital, Stuttgart, Germany
4
Hematopathology Unit, Pathology Department, Hospital Clínic and University of Barcelona, Institute
of Biomedical Research August Pi i Sunyer (IDIBAPS), Barcelona, Spain
5
Institute of Pathology, University of Würzburg, Würzburg, Germany and Comprehensive Cancer
Center Mainfranken (CCCM)
6
Institute of Pathology, Caritas-Hospital, Bad Mergentheim, Germany
We have recently shown that immunoblastic (IB) morphology in diffuse large B-cell
lymphoma (DLBCL) indicates a significant risk for shorter survival (Ott et al. 2010,
Blood). At the same time, the presence of MYC rearrangements in DLBCL has been
shown to indicate shorter survival in R-CHOP treated patients. Since we repeatedly
detected the presence of a MYC rearrangement in DLBCL with IB morphology in our
daily diagnostic experience we conducted a systematic study to determine the frequency of 8q24/MYC rearrangements in DLBCL of immunoblastic type (IB-DLBCL)
versus data obtained from a group of non-IB DLBCL.
Paraffin-embedded tumor samples of 108 DLBCLs, 39 of them with IB- and 69 with
non-IB morphology were investigated by fluorescence in situ hybridization (FISH)
with a break-apart probe to detect a translocation of the MYC-gene and by immunohistochemistry for protein expression of CD10, MUM1/IRF4, BCL2, BCL6, Ki67, and
MYC. Additionally, cases with a signal constellation indicating MYC-translocation
were hybridized with break-apart probes for BCL2, and BCL6, as well as with a
MYC-IgH fusion probe.
Signal constellations indicative of a translocated MYC gene were found in 13/39 IB
DLBCL (33.3%). In contrast, in the non-IB DLBCL samples only 5/69 (7.2%) MYC rearrangements (MYCR) were observed (p<0.01). 12/13 MYCR IB-DLBCL harbored a
fusion with the immunoglobulin heavy chain gene (MYC-IgH), while one sample was
not interpretable. Moreover, MYCR was observed in association with BCL6 breakage
in 2/13 (15.4%), with a dual BCL2 and BCL6 breakage in 1/13 (7.7%), and with an
amplification of the BCL2 gene (1/13; 7.7%).
Interestingly, MYCR IB-DLBCL samples showed CD10 positivity more often
(8/13; 62%) in contrast to their MYC non-rearranged counterparts [4/26 (15%);
p<0.01]. A significantly higher proportion of MYC break-positive IB had a Ki67
index >80% in comparison to MYC break-negative tumors [11/13 (84.6%) vs.
13/26 (50%), p<0.05]. 16 IB-DLBCLs showed strong overexpression (>= 80%
cells) of MYC protein (MYChigh). In the MYCR group 11/13 (85%) cases were
MYChigh whereas only 5/26 (19%) were MYChigh in the translocation-negative
group [p<0.01]. No significant differences regarding protein expression of BCL2,
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BCL6, and IRF4/MUM1 between the MYC translocated and the MYC germline
cases were observed.
In conclusion, MYC-IgH rearrangements frequently occur in IB-DLBCL without additional BCL2- and/or BCL6-translocations. The activation of MYC, therefore, may be
an important pathogenic feature in IB-DLBCL.
Keywords: MYC; Immunoblastic B-Cell Lymphoma; FISH
[OP-LYMP-19]
CD23-POSITIVE DIFFUSE NODAL FOLLICULAR LYMPHOMA:
A DISTINCT VARIANT OF FOLLICULAR LYMPHOMA
MIMICKING NODAL MARGINAL ZONE LYMPHOMA
Keegan Barry Holson1, Charles Ma2, Lizalynn Dias2,
Jane Houldsworth2, Russell K Brynes1, Imran N. Siddiqi1
1
Department of Pathology, University of Southern California Keck School of Medicine, Los Angeles,
California, USA
2
Cancer Genetics, Inc., Rutherford, New Jersey, USA
CD23 co-expression and diffuse/ interfollicular growth pattern are uncommon
findings in nodal follicular lymphoma (FL) and can create diagnostic challenges. Similar cases are rarely described in the literature, most notably by
Katzenberger et al. (Blood, 2009) who proposed a unique FL variant, characterized by diffuse growth pattern, lack of IGH/BCL2 translocation, presence of
1p36 deletion, frequent inguinal lymph node involvement, CD23 co-expression, and low clinical stage. Other studies on CD23+ FL, while associating
inguinal location, have not specifically described this architecture. In addition, no follow-up studies have correlated pathologic findings with clinical and
genetic features, and these cases remain a diagnostic problem, mimicking
marginal zone lymphoma, small lymphocytic lymphoma or rarely even reactive
hyperplasias.
Figure 1. CD23+ diffuse follicular lymphoma (case 2). A diffuse/interfollicular
growth pattern of lymphoma is observed, focally surrounding well-preserved
lymphoid follicles, with reactive germinal centers and intact mantle zones (A,
H&E x40) or, more commonly, surrounding very ill-defined and partially colonized
germinal centers (B, H&E x40). The lymphoma (diffuse/ interfollicular component)
is composed of mixed centrocytes and centroblasts (A, inset, H&E x1000), and is
diffusely positive for CD20 (C, x40) and CD23 (D, x40). CD21 highlights follicular
dendritic cell networks associated with rare residual follicles, while the lymphoma
is devoid of follicular dendritic cell networks (E, x40). CD10 and BCL-6 stain
residual germinal centers, with more variable staining of the lymphoma (F and
G, respectively, x40). BCL2 is positive in the lymphoma and negatively highlights
residual germinal centers (H, x40). Ki67 demonstrates high proliferation limited to
residual germinal centers, with lower proliferation in the lymphoma (I, x40).
Ten cases of diffuse, CD23+ FL were identified along with pertinent clinical
information. Cases were analyzed by histomorphology, immunohistochemistry
(including CD3, CD10, CD20, CD21, CD23, CD43, BCL2, BCL6, LMO2, Ki67), FISH
(IGH/BCL2 translocation), and array CGH on FFPE tissues (MatBATMarray, Cancer
Genetics, Inc.).
All cases were CD23+, and 8 of 10 involved inguinal lymph nodes. Patient ages
ranged from 35 to 89 (mean 56.7 years), without a gender predilection. Three
patients were low stage (I-II), while six presented with high stage disease (IIIIV). Histologically (Figure 1), a predominantly diffuse and interfollicular pattern of
mixed centrocytic/centroblastic cells was evident. Residual lymphoid follicles with
reactive-appearing germinal centers were present in all cases; most often, these
were extremely ill-defined, partially colonized and only appreciable by immunohistochemistry (CD10+, Bcl-6+, Bcl-2-, high Ki67), but in some cases they were
histologically intact with preserved mantle zones. Neoplastic B-cells diffusely surrounded the remnant follicles and were uniformly CD20+, CD23+, Bcl-2+, and
CD43-, with variable Bcl-6 expression. CD10 was at least partially expressed in 7
of 10 cases. In the three CD10- cases, distinction from marginal zone lymphoma
was particularly difficult, but the cytological features and Bcl-6 and/or LMO2
expression were helpful for classification as FL. Relative proportions of centrocytes/ centroblasts (i.e. cytologic grade) did not correlate with immunophenotype
or clinical stage. By FISH analysis, the vast majority of cases lacked IGH/BCL2
translocation (7 of 9 cases tested). However, genomic gains/losses were detected
in 8 of 9 cases analyzed by array CGH (Figure 2). Deletion of 1p36 was observed
in 4 cases and included TNFRSF14, 2 with concurrent IGH/BCL2 translocation.
Other recurrent aberrations included loss of 4q34-q35 and gains of 3p, 3q, and/
or 12q, changes more frequently reported in nodal marginal zone lymphoma than
in conventional FL. Gains of 2p (including focal gains of REL), 9q and 11p were
also recurrently observed.
Figure 2. Ideogram showing loci of chromosomal gains/losses. DNA was
extracted from 5x10 micron sections of each specimen, and when necessary
fragmented to 400-1200bp prior to labeling. Reference DNA (equimixture of control
male:female DNA) was similarly fragmented and labeled. Labeled DNAs were
hybridized to a custom-designed microarray targeted to represent genomic regions
commonly altered in mature B-cell neoplasms. Nexus (Version 7.0, Biodiscovery,
Inc.) was used for aberration detection and visualization. Blue bars denote gains,
red bars denote losses, and each bar represents one case. Of note, some focal
gains/losses were identified as common CNVs.
CD23+ diffuse FL has a characteristic histopathology and immunophenotype,
frequently involves the inguinal region, is often negative for IGH/BCL2 translocation, and displays patterns of genomic gain and loss dissimilar to conventional FL, features supportive of a distinct clinicopathologic variant. The peculiar
diffuse/interfollicular growth pattern is important to recognize, as it can mimic
other small B-cell lymphomas, nodal marginal zone lymphoma in particular, and
may have clinical relevance.
Keywords: Diffuse follicular lymphoma, BCL2/IGH negative follicular lymphoma, array CGH
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89/f
35/f
40/m
63/m
62/m
51/f
5
6
7
8
9
10
NOVEL MARKERS FOR THE DIAGNOSIS OF PAEDIATRIC
FOLLICULAR LYMPHOMA
Ayse U. Akarca1, Hasan Rizvi2, Claudio Agostinelli3, Alan Ramsay4,
Maria Pane Foix4, Joan Somja4, James Wilton1, Vishvesh H. Shende1,
Brunangelo Falini5, Stefano A. Pileri3, David Linch6,
Stephen Daw7, Teresa Marafioti8
1
Department of Pathology, University College London, UK
Department of Cellular Pathology, Barts Health NHS Trust, London, UK
3
Section of Haematopathology, Department of Haematology and Oncological Sciences “Seràgnoli”,
S. Orsola-Malpighi Hospital, University of Bologna, Italy
4
Department of Cellular Pathology, University College Hospital London, UK
5
6
7
Children and Young People’s Cancer Service, University College Hospital London, UK
8
NIHR UCLH/UCL Biomedical Research Centre London, UK ‘;’ Department of Pathology, University
College, London, UK
2
Paediatric follicular lymphoma (PFL) accounts for 1-2% of all paediatric lymphomas. It occurs under 18 yrs of age and presents as localized disease. The prognosis is generally excellent. PFL shows distinct features from adult FL. The follicles
contain numerous blast-like cells with a “starry-sky” appearance correlating with
high proliferation and histological grade. The distinction between PFL and follicular
hyperplasia (FH) is sometimes difficult. PFLs usually lack t(14;18) translocation are
BCL-2-negative and often IRF-4/MUM1-positive. IgH/IRF4/MUM1 translocation and
1p36 deletion are recurrent events. We undertook a histopathological review of 15
PFLs and assessed by immunohistochemistry the diagnostic value of molecules like
Stathmin, FOXP-1 and TNFRSF14/HVEM and their utility to distinguish PFL from FH.
Paraffin-blocks were obtained from the Department of Pathology at UCLH. The diagnosis was performed by expert haematopathologists. Clinical-pathological data are
summarised in Table 1 and 2. The normal architecture was replaced by a nodular
proliferation of neoplastic cells. Three patterns were observed: a) large follicles filled
with centroblasts, scattered macrophages with a “starry-sky” appearance and thin
mantle-zone; b) irregularly-shaped sometimes coalescent follicles with centroblasts
and centrocytes and ill-defined mantle-zone; c) follicles with variable number of
centroblasts with a peripheral rim of marginal zone-like B-cells extending into the
extrafollicular area. Immunohistochemistry highlighted preserved BCL-6 and CD10
positivity (13/13 and 14/14 respectively); predominance of K light-chain restriction (9/12=75%) and lack of BCL-2 (clones 124 and E17) expression (12/15=80%).
Stathmin and a new GC-associated protein (called GACP) were widely positive
(11/11=100% and 10/11=91% respectively) as was FOXP-1 (10/11=91%) labelling scattered cells in reactive GC. IRF-4/MUM-1 was found in 3/14 =21% cases.
Additionally, 6 cases revealed IRF-4/MUM-1 expression in a proportion of neoplastic
cells with marginal zone differentiation. Similarly, 2 out of 8 investigated cases were
IRTA-1-positive (the 2 cases were IRF-4/MUM-1-positive). Furthermore, 4 samples
showed IRTA-1 in neoplastic cells at the periphery of GCs. TNFRSF14/HVEM was detectable in only 2/11 (18%) samples with 2 additional cases highlighting occasional
neoplastic cells in a few follicles. TNFRSF14/HVEM gene is located in 1p36, a region
frequently lost in PFLs; nonsense-mutations in TNFRSF14/HVEM have been detected in 41% of PFLs. FISH revealed no evidence of bcl-2 an/or bcl-6 rearrangements
in the examined cases (0/12) with the exception of 4 samples in which increased
copy numbers were seen. By PCR, Ig genes rearrangement were detected in 67%
(6/9) of PFLs. In summary, we found that a) PFLs present different histopathological
patterns; b) Stathmin, GACP and FOXP-1 merit inclusion in the diagnostic panel; c)
FOXP-1, IRF-4/MUM-1 and IRTA-1 are useful to differentiate PFLs from FH (usually
negative) and valuable to identify PFLs with marginal zone differentiation; d) lack of
TNFRSF14/HVEM expression suggests it as a possible surrogate of the 1p36 deletion commonly seen in PFLs. Whether, other chromosomal alterations involving increased copy numbers of bcl-2, bcl-6 and FOXP-1 alone or together with the already
described IgH/IRF-4/MUM-1 rearrangement and 1p36 del are the genetic events
responsible for the malignant transformation is under investigation.
Lymph node site: I = inguinal; C = cervical, SC = supraclavicular; A = axillary; M = mediastinal; Ab = abdominal/ pelvic; SM = submandibular; P = parotid; R = retroperitoneal; EN = extranodal Residual germinal centers: WD = well-defined (intact germinal centers and mantle zones); ID = ill-defined (no mantle zones, partially
colonized germinal centers). N/A = not available
Gain: 1q, 11p, 12q
Loss: 6q, 11q
Focal WD, ID
61/f
4
I, SM
IA
3
+
+
+
N/A
-
N/A
-
Gain: 9q (partial)
N/A
+
+
+
+
+
1
2
ID
Focal WD, ID
IIB
61/f
3
I, Ab, R
IIIB
I, C, A, M
+
N/A
NO CHANGES
ID
IA
I
2
+
+
+
N/A
-
Gain: 3q (partial)
ID
IIIB
C, A, SC, Ab, EN
(breast)
2
+
+
+
+
-
Gain: 2p (focal), 3, 6p. Loss:
4q (partial), 8q (focal), 10p
(focal), 10q (focal), 20q (focal)
+
focal ID
N/A
I
2
+
+
+
+
-
Gain: 9q
Loss: 9p
+
focal ID
IVB
C, A, M, Ab
2
+
-
+
-
-
Gain: 2p (focal), 11p, 12q
Loss: 4q (partial)
+
+
+
+
55/m
2
ID
IIIA
I, A, M, Ab
2
+/+
+
focal WD, ID
IIIA
I, C, P
3
+
+
Gain: 2p (focal), 12q, 13q
Partial
+
Gain: 2p, 3, 12
-
+
1p36 deletion
+
+
Bcl-6
CD10
+
CD23
ID
IVA
Cytologic Grade
Residual Germinal Centers
Stage
Site
50/m
1
I, SC, A
Age/sex
Case
Table 1. Summary of CD23+ diffuse follicular lymphoma cases
1
LMO2
BCL2/IGH FISH
Other Gains and Losses
[OP-LYMP-20]
Keywords: Pediatric Follicular Lymphoma, BCL-2 negative, Stathmin
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[OP-LYMP-21]
THE ROLE OF AKT/MTOR CASCADE IN HAIRY CELL LEUKAEMIA:
INTERACTION WITH BRAF SIGNALLING PATHWAY AND
PROGNOSTIC SIGNIFICANCE
Georgia Levidou1, Eleftheria Lakiotaki1, Maria K. Angelopoulou2,
Gerasimos A. Pangalis3, Theodoros Vassilakopoulos2,
Angelica A. Saetta1, Athanasia Sepsa1, Ilenia Chatziandreou1,
Gabriella Gainaru2, Pagona Flevari2, Sotirios Sachanas3,
Maria Moschogiannis3, Christina Kalpadakis4, Vasilis Milionis1,
Panayiotis Panayiotidis5, Maria Dimopoulou2, Eleni Plata2,
Konstantinos Konstantopoulos2, Efstratios Patsouris1,
Penelope Korkolopoulou1
1
Department of Hematology and Bone Marrow Transplantation, University of Athens, Medical
School, Greece
3
Department of Hematology, Athens Medical Center, Psychikon Branch, Greece
4
Department of Hematology, University of Crete, Heraklion, Greece
5
1st Department of Propedeutic Internal Medicine, University of Athens, Medical School, Greece
2
Background: The molecular hallmark of Hairy cell leukaemia (HCL) is BRAFV600E
mutation, which results in constitutive activation of the corresponding signalling
pathway. In this study we focused on any possible interrelations between BRAF and
several key molecules belonging to Raf-MEK-ERK and AKT/mTOR pathway as well
as on their associations with clinicopathological and bone marrow microvascular
characteristics.
Methods: 77 patients with HCL for whom bone marrow biopsy at diagnosis and
clinical data were available were enrolled in this investigation. Strict selection criteria were used for patient inclusion. All patients had been followed-up for a median
of 9.97 years and the majority of them had been treated with IFN-. All cases were
investigated immunohistochemically for the expression of phosphorylated (p-) AKT,
p-m-TOR, BRAFV600E, p-ERK1/2, p-p70S6K and p-4EB-P1. Information regarding
the microvascular characteristics highlighted by anti-CD34 was available in 36 cases. The presence of BRAFV600E mutation was investigated by HRMA in 35 cases.
th
Table 1. Clinical pathological data of 15 paediaric follicular lymphomas
Table 2. Age and sex distribution in 15 cases of paediatric follicular
lymphoma
Results: p-mTOR expression was present only in 10/77 cases. Cytoplasmic pp70S6K and p-4E-BP1 immunoexpression was seen in 72/77 and 75/77 cases.
The 10 cases being positive for p-mTOR coexpressed p-p70S6K and p-4E-BP1,
thus denoting that full activation mTOR cascade does occur in HCL, but appears
uncommon. 59 cases being p-p70S6K(+) but p-mTOR(-) were p-ERK1/2(+), consistent with activation of 4EBP1 directly by ERK. All 65 cytoplasmic p-4E-BP1 positive
cases which were p-mTOR(-) displayed cytoplasmic p-AKT, indicating that AKT may
activate p70S6K independently from m-TOR. p-AKT immunoreactivity was observed
in 96.1% cases. p-ERK1/2 cytoplasmic expression was recorded in all cases, with
18 cases displaying also nuclear expression. BRAFV600E immunoreactivity was
present in all cases. BRAFV600E mutation was detected in 30/35 cases. p-ERK1/2
was positively correlated with p-mTOR, cytoplasmic p-p70S6K and cytoplasmic
p-4E-BP1, corroborating the well-known interactions between these two pathways.
Regarding microvascular characteristics only nuclear p-ERK1/2 expression was
positively correlated with shape factor of bone marrow microvessels. In univariate
survival analysis increased p-AKT, cytoplasmic p-ERK1/2, cytoplasmic p-4E-BP1,
p-mTOR and BRAFV600E were correlated with shorter time to second treatment.
The same applied to the presence of simultaneous overexpression of p-AKT/cytoplasmic p-mTOR/ cytoplasmic p-4E-BP. In multivariate survival analysis the combination of p-AKT/p-mTOR/p-4E-BP1 coexpression was proven to be an independent
predictor of adverse prognosis, along with degree of bone marrow infiltration by
hairy cells and the presence of hepatomegaly. When p-AKT, p-mTOR and p-4E-BP1
were entered separately in Cox’s model, only the latter emerged as the independent
prognosticator.
Conclusion: Our results confirm the presence of BRAFV600E mutant protein and the
consequent MAPK pathway activation culminating in ERK phosphorylation. We also
provide evidence for the first time about the activation of AKT/mTOR pathway in a
small subset of HCL cases and illustrate the interaction between the two pathways.
mTOR downstream targets, p-p70S6K and p-4E-BP1 seem to be constitutively expressed obviously due to independent inputs from AKT and MAPK. p-4E-BP1, on its
own or combined with p-AKT and p-mTOR is brought forward as an independent
adverse prognosticator of time to second treatment.
Keywords: Hairy cell leukemia, BRAFV600E, AKT/mTOR pathway
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| 17-22 October 2014
Julia Gonzalez Rincon1, Sagrario Gomez1, Carol Lozano1,
Miguel Piris Villaespesa1, Marcos Gonzalez2, Eduardo Anguita3,
Rosa Collado4, Felix Carbonell4, Raul Cordoba5, Antonia Rodriguez6,
Nerea Martinez7, Miguel Ángel Piris8, José Antonio Garcia Marco9,
Margarita Sanchez Beato1
1
Instituto Investigación Sanitaria Puerta de Hierro, Madrid, Spain
Hospital Universitario Salamanca-Centro Investigación Cancer, Salamanca, Spain
Hospital Clinico de Madrid, Spain
4
Consorcio Hospital General Universitario de Valencia, Spain
5
Hospital Universitario Infanta Sofia, Madrid, Spain
6
Hospital Universitario 12 de Octubre, Madrid, Spain
7
Instituto de Investigación Marqués de Valdecilla, Santander, Spain
8
Hospital Universitario Marques de Valdecilla- Instituto de Investigación Marqués de Valdecilla,
Santander, Spain
9
Hospital Universitario Puerta de Hierro-Majadahonda, Instituto Investigación Sanitaria Puerta de
Hierro,Madrid, Spain
2
3
Background: Chronic Lymphocytic Leukemia (CLL) is the most common adult leukemia in the western world. Although the mutational status of IgHv and of TP53 have
been the most relevant prognostic molecular markers in CLL for a long time, next
generation sequencing data are revealing new genes whose alterations are relevant
for the pathogenesis and progression of CLL. Although an important number of studies have been done with NOTCH1, SF3B1, BIRC3 and others, little is known about
the impact and frequency of other genes found recurrently mutated in CLL. Our aim
was to study the frequency, association and impact of these new genetic markers
mutations in untreated progressed CLL patients.
Design: We have done massive parallel sequencing after target enrichment with
HaloPlex system from Agilent. A total of 25 genes, previously found as mutated in
CLL were included in the design (NOTCH1, SF3B1, TP53, ATM, BIRC3, MYD88, XPO1,
POT1 and others). We have analyzed 100 samples from CLL patients with clinical
and molecular data. Eighty of these 100 patients are enrolled in the REM study evaluating first-line therapy with Rituximab, fludarabine and cyclophosphamide (RFC).
Results: The most frequently mutated genes were SF3B1, ATM, NOTCH1, LRP1B and
CSMD3 in around 20%, 15%, 13%, 10% and 9%, respectively. Other genes mutated in
2 or more samples were POT1, TP53, SI, PLEKHG5, BIRC3, FBXW7, BRAF, CHD2, KLHL6,
MYD88, NFKBIE, SAMHD1, TGM7 and ZMYM3. Around 50% of the analyzed samples
showed 2 or more mutations, whereas 24% showed no mutation in any of the genes.
Methods: Patients with mantle cell lymphoma were identified via archive search of
institutional databases. Clinical information was collected from electronic medical
records. Genomic DNA was extracted from formalin-fixed paraffin embedded tissue using a kit (Qiagen). IGHV mutation assay was performed using IGH Somatic
Hypermutation Assay v2.0 (InVivoScribe Technologies) per manufacturer’s instructions. DNA sequence data was compared to published germline sequence using
IgBLAST (http://www.ncbi.nlm.nih.gov/igblast/). Cases were categorized as unmutated (UN, 100% identity), minimally/borderline mutated (MIN, 99.9%-97%) or significantly mutated (SIG, 96.9-95%). Immunohistochemical stains for Sox11 (clone
MRQ-58, CellMarque, Rocklin, CA) and Ki67 (clone 30-9, Ventana, Tucson, AZ) were
performed on automated stainers and nuclear staining was visually scored. This
study was approved by local Institutional Review Boards.
Results: A total of 47 cases were sequenced. One case (2% of all cases) was UN, 36
(77%) were MIN, and 10 (21%) were SIG. There was no significant difference in age
between the groups. Overall survival data was available for the one UN case (alive at
119 months), 34 MIN cases, and 6 SIG cases. Although the MIN cases had a longer
median survival (41.5 months) than the SIG cases (30.5 months) the difference was
not significant (p=0.74). Survival analysis with log-rank test (figure 1, MIN=red line,
SIG =green line) also did not show a significant difference in survival (log-rank test
p=0.238) although there was a trend for worse survival in the SIG group.
26 MIN and 4 SIG cases had tissue available for SOX11 IHC; four MIN cases lacked
expression of SOX11 (less than 30% of tumor cells positive) and all SIG cases were
positive. There was no association between SOX11 positivity and mutation status
(Fisher’s exact test p=1.00). 20 MIN and 4 SIG cases had material for Ki67 staining. There was no significant difference in % Ki67-positive cells between the two
groups (p=0.21).
RECURRENT GENE MUTATIONS IN CHRONIC LYMPHOCYTIC
LEUKEMIA: RESULTS FROM THE REM CLINICAL TRIAL
suggested that MCL which have undergone significant mutation of IGHV genes
(<97% identity) may correspond to those having a more indolent course with a nonnodal presentation and negativity for SOX11. We investigated these associations in
our cohort of MCL cases from three different institutions.
[OP-LYMP-22]
MIPI scores were available for 21 MIN cases and 2 SIG cases; there was no significant difference in MIPI between these two groups (p=0.11). Across all patients
with MIPI data available, there was a significantly worse survival for patients with
a high risk score (>6.1) compared to low and intermediate risk patients (log-rank
test p=0.03).
Conclusions: Our current data does not confirm recent findings that MCL with significant IGHV mutation are associated with more indolent behavior or lack of Sox11
expression. The proportion of MIN cases (77%) was higher than has previously been
reported. Thus analysis is limited by the small numbers of UN and SIG patients.
Keywords: Mantle Cell Lymphoma, IGHV, SOX11
ATM and TP53 seem to be mutually exclusive and, although SF3B1 and NOTCH1
tend to be also mutually exclusive, two samples showed mutation in both genes.
There are no clear concurrent or exclusive mutations for the rest of the genes.
Associations with clinical, molecular and laboratory characteristics are currently being analyzed.
Conclusion: In our group of untreated, progressed CLL patients, SF3B1, followed by
ATM, NOTCH1, LRP1B and CSMD3, are the most frequently mutated genes, showing
frequencies from 9 to 20%. The impact of these mutations on the clinical course is
currently being analyzed.
Keywords: Chronic Lymphocytic Leukemia, massive sequencing, gene mutation
[OP-LYMP-23]
IGHV MUTATIONAL STATUS AND POTENTIAL ASSOCIATIONS
WITH SOX11 EXPRESSION AND SURVIVAL IN MANTLE CELL
LYMPHOMA
Megan O. Nakashima1, Xiaoxian Zhao1, Xianqian Li1, Lisa Durkin1,
Brian T. Hill2, Kai Fu3, Raymond Lai4, Eric D. Hsi1
Figure 1. Survival curves for minimally/borderline mutated cases (red) and
significantly mutated cases (green).
1
Laboratory Medicine, Cleveland Clinic, Cleveland, USA
Hematology Oncology, Cleveland Clinic, Cleveland, USA
3
Pathology and Microbiology, University of Nebraska, Omaha, USA
4
Laboratory Medicine and Pathology, University of Alberta, Edmonton, Canada
2
Introduction: Mantle cell lymphoma (MCL) is characterized by t(11;14) and usually
has an aggressive course, although occasional cases behave indolently. Published
data is somewhat conflicting regarding whether degree of somatic IGHV mutation is
associated with prognosis. However a recent study (Navarro et al, Cancer Res 2012)
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th
[OP-LYMP-24]
DETECTION OF MYD88 L265P AND CXCR4 MUTATIONS IS A
VALUABLE TOOL FOR DIAGNOSIS OF LYMPHOPLASMACYTIC
LYMPHOMA AND IDENTIFIES CASES WITH HIGH DISEASE
ACTIVITY
Janine Schmidt, Natalie Schindler, Irina Bonzheim,
Birgit Federmann, Falko Fend, Leticia Quintanilla Martinez
Institute of Pathology and Neuropathology, University Hospital Tübingen, Tübingen, Germany
Background: Lymphoplasmacytic lymphoma (LPL) is a small B-cell lymphoma usually presenting with bone marrow (BM) involvement and serum monoclonal IgM
protein, also termed Waldenström’s macroglobulinemia. Differential diagnosis from
other small B-cell lymphomas can be difficult due to lack of specific immunophenotype. Recurrent mutations in MYD88 have been identified in >90% of LPL cases but
are rare or absent in other small B-cell lymphomas. Recently, WHIM syndrome-like
mutations in CXCR4 have been identified in a fraction of LPL cases, and seem to impact clinical presentation and response to therapy. We investigated the presence of
MYD88 L265P and CXCR4 mutations in decalcified paraffin-embedded BM biopsies.
Additionally, we investigated potential differences in clinical presentation between
CXCR4WHIM vs. CXCR4WT cases.
Design: 90 decalcified FFPE BM biopsies were analyzed including 51 cases of LPL,
14 cases of B-cell chronic lymphocytic leukemia (CLL), 13 cases of marginal zone
lymphoma (MZL) and 12 normal BM samples. For identification of MYD88 L265P a
LNA-clamped PCR with subsequent melting curve analysis was developed. A subset
of cases was validated with Sanger sequencing. In addition, the C-terminal domain
of CXCR4 was sequenced in LPL cases.
Results: The median age of patients with LPL was 71 years (range 46 – 89) with a
slight male predominance. The mean IgM level at diagnosis was 2952 mg/dl (range
95 – 7800). The extent of BM infiltration ranged from 5 – 100% (mean 42.5%).
Only one case showed an IgG paraprotein. The age and sex ratio at presentation
was similar among LPL, MZL and CLL patients. However, patients with CLL tended
to have higher BM tumor burden (mean 62.5%) followed by LPL (mean 42.5%)
and MZL (mean 35%) cases. MYD88 L265P was found in 49/51 (96%) LPL cases,
and 1/13 (7.7%) splenic MZL, whereas all CLL samples and reactive controls remained negative. The two MYD88WT LPL cases had low BM infiltration, low IgM
levels and a history of cold agglutinin disease (CAD) associated to autoimmune hemolytic anemia. Mutations in CXCR4 were detected in 20/46 (43.5%) analyzed LPL
cases. CXCR4WHIM mutated LPL cases showed a significantly higher BM infiltration
(p=0.023), and lower counts of hemoglobin (p=0.008), platelets (p=0.016) and leukocytes (p=0.029). In contrast, CXCR4WT cases tended to have higher incidence of
lymphadenopathy (41% vs. 22%). CXCR4nonsense mutated cases showed significantly
lower platelet counts and higher IgM levels at diagnosis, whereas CXCR4frameshift mutated cases had a significantly lower hemoglobin level when compared to CXCR4WT
cases.
Conclusions: 1) LNA-clamped PCR with melting curve analysis is a fast and reliable
method for detecting the MYD88 L265P mutation in FFPE samples and is useful for
subtyping small B-cell lymphomas in BM biopsies. 2) We confirm that LPL patients
with cold agglutinin disease are negative for the MYD88 L265P mutation suggesting
a separate disease entity. 3) The incidence of CXCR4 mutation in our study is higher
than previously described and might reflect a selection towards cases with higher
tumor burden in BM biopsies, as compared to peripheral blood and BM aspirates.
4) CXCR4 mutations identify a subgroup of LPL patients with higher disease activity.
Keywords: LPL, MYD88 L265P, CXCR4
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POSTER PRESENTATIONS
Table 1. The value of PDGFRβ score for the identification of progressive
bone marrow fibrosis. Specificity and sensitivity was calculated considering all
evaluated cases for the full length of follow-up, for the short-term follow-up and
separately for pre-fibrotic cases only. End-stage (MF-3) fibrosis was excluded
as these cases were no subjects of further progression. TP: true positive, TN:
true negative, FP: false positive, FN: false negative.
IMMUNOHISTOLOGICAL EVALUATION OF PDGFRSS
EXPRESSION IN BONE MARROW IS POTENTIALLY
PREDICTIVE FOR FIBROSIS PROGRESSION IN PREFIBROTIC
MYELOPROLIFERATIVE NEOPLASIA: AN EUROPEAN BONE
MARROW WORKING GROUP STUDY
Gábor Méhes1, Alexandar Tzankov2, Konnie Hebeda3,
Ioannis Anagnostopoulos4, László Krenács5, Judit Bedekovics1
1
Department of Pathology, University of Debrecen Clinical Centre, Debrecen, Hungary
Institute of Pathology, University Hospital, Basel, Switzerland
Department of Pathology, Radbound University Nijmegen Medical Centre, Nijmegen, The
Netherlands
4
Institute of Pathology, Charité-University Medicine Berlin, Campus Charité Mitte, Berlin, Germany
5
Laboratory of Tumor Pathology and Molecular Diagnostics, Szeged, Hungary
2
3
Aim: Myelofibrosis is the result of stromal reaction of the bone marrow which is
measured histologically by the reticulin staining in the daily practice. As fibroblast
activation is required for fiber synthesis, the aim of the study was to evaluate the
amount of cellular activation in progressive myelofibrosis.
Methods: 193 initial and follow-up bone marrow biopsy samples from 84 patients
with myeloproliferative neoplasia were provided by five hematopathology centers.
Estimation of the fiber mass (MF grade, 0-3) by reticulin silver staining and fiber producing PDGFR positive stromal cells (PDGFR score, 0-3) by immunohistochemistry were performed in each sample and correlated for prediction of progression.
Results: Strong correlation between the MF grade and PDGFR score (Spearman
r was 0.83, p<0.0001) could be verified. In general, PDGFR scores higher than
MF grade predicted the myelofibrosis progression with 43% sensitivity and 57%
specificity. The short-term analysis resulted in 82% sensitivity but only 53% specificity. Most interestingly, pre-fibrotic cases (MF-0) with elevated PDGFR expression
turned out to be progressive (90% sensitivity and 75% specificity).
Conclusions: In summary, PDGFR highlights the cellular aspects of marrow fibrosis in close relation with stromal matrix accumulation. Our data indicate to a direct
prognostic impact in prefibrotic myeloproliferative disorders.
The work was supported by the TÁMOP- 4.2.2.A-11/1/KOV-2012- 0045 “Research
network on vascular biology/medicine” project grant.
Keywords: Bone Marrow, Primary Myelofibrosis, Platelet-Derived Growth Factor
Receptor
[PP-BM-02]
PROGNOSTIC SIGNIFICANCE OF THE IGVH MUTATION
STATUS AND IMMUNOHISTOCHEMICAL ANALYSIS OF ZAP70
AND NFK-B IN BONE MARROW BIOPSIES IN CHRONIC
LYMPHOCYTIC LEUKEMIA
Furuzan Kacar Doger1, Ibrahim Meteoglu1, Nesibe Kahraman Cetin2,
Gokhan Pektas1, Irfan Yavasoglu1
1
2
P OS T ER P RES EN TAT I ON S
[PP-BM-01]
Department of Pathology, Faculty of Medicine, Adnan Menderes University, Aydın, Turkey
Yozgat Government Hospital, Yozgat, Turkey
Objective: Chronic lymphocytic leukemia (CLL) is one of the most common types of
leukemias among adults in industrialized countries. It is important to recognize patients with a more rapid course of disease due to the nature of CLL. The goal of our
study is to validate ZAP70 and NFK-B antibodies along with İmmunglobulin heavy
chain variable region (IgVH) mutation status which have been associated with rapid
progression and aggressive clinical course in CLL and to correlate the expression of
these molecules with patterns of bone marrow infiltration.
Materials-Methods: We have included 84 bone marrow biopsy samples into
the study to determine ZAP70 and NFK-B status using immunohistochemistry.
Expressionpatterns for both antibodies were then correlated with the bone marrow infiltration patterns. We have also analyzed by IgVH mutational status from 20
patients using DNA obtained from paraffin embedded formalin fixed bone marrow
biopsies. These findings were then correlated with immunohistochemical results.
Results: We have identified a positive correlation between the expression patterns
of ZAP70 and NFK-B, factors that have been previously identified as poor prognostic
factors(p<0.001). However there was no correlation between these two markers
and the IgVH mutation status in these patients (p=1.000, p=0.931). In addition, we
have shown a statistically significant oppositive correlation with ZAP70 immunostaining and the necessity for early intervention (p=0.046).There was a statistically
significant ZAP70 and NFK-B expressions in the diffuse pattern of bone marrow
infiltration. (p<0.001 and p<0.001, respectively).
Figure 1. Microscopic images of follow-up samples from a case diagnosed with
PMF. Hypercellularity (H&E staining), no fiber accumulation (MF-0, reticulin staining) and
a mild stromal cell activation (PDGFRβ score-1, immunohistochemistry) was visible in
the initial biopsy sample (upper row). Same hypercellularity with mild increase of fibrosis
(MF-1) accompanied with extended stromal cell activation (PDGFRβ score-2) highlighted
in the follow-up sample obtained seven months later (lower row).
Conclusion: Despite the limited numbers, the findings of our study suggests that
ZAP70 and CD38 expression patterns as well as IgVH mutation status might be
helpful to determinethe course of the disease and risk of progression. Specifically
ZAP70 immuno positive patients appear to have a faster progression of their disease and may require earlier intervention and closer follow up.
Key Words: Chronic lypmhocytic leukemia, IgVH mutation, ZAP70, NFK-B
Keywords: leukemia, IgVH, NFK-B
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[PP-BM-03]
A COMPARISON BETWEEN IMMUNOFIXATION
ELECTROPHORESIS, MULTICOLOR FLOW CYTOMETRY AND
IMMUNOHISTOCHEMISTRY IN DETECTING MINIMAL RESIDUAL
DISEASE IN PLASMA CELL MYELOMA
Tariq N. Aladily1, Xiaohong I. Wang2, Beverly Handy3,
Denai R. Delton4, Pei Lin2
1
Department of Hematopathology, The University of Texas-MD Anderson Cancer Center, Houston,
Texas, USA; Department of Pathology, The University of Jordan, Amman, Jordan
Department of Hematopathology, The University of Texas-MD Anderson Cancer Center, Houston,
Texas, USA
3
Department of Laboratory Medicine, The University of Texas-MD Anderson Cancer Center,
Houston, Texas, USA
4
Department of Biostatistics, The University of Texas-MD Anderson Cancer Center, Houston, Texas,
USA
2
Objectives: To compare sensitivity of different testing methods for detection of minimal
residual diseases (MRD) in plasma cell myeloma.
Methods: We analyzed the results of immunofixation electrophoresis (IFE), multicolor
flow cytometry (FC) acquiring 100K events and immunohistochemistry/ in situ hybridization for immunoglobulin light chain and CD138 (IHC/ISH) from 100 consecutive cases of myeloma patients after therapy between June and September 2011.
Treatment modalities included induction chemotherapy or stem cell transplant. Only
cases with plasma cell count <= 5% in bone marrow aspirate were included in the
study. Residual disease was diagnosed if FC and/ or IHC demonstrated monoclonal
and/ or aberrant plasma cells phenotype, or if a positive result of IFE persisted for more
than four weeks. Flow cytometry analysis is based on a panel of antibodies including,
CD19, CD20, CD28, CD45, CD38, CD117, CD138, cytoplasmic K and cytoplasmic L,
acquiring 100K events. IHC was performed on the biopsy or/and clot sections.
Results: The percentage of PCs on aspirate smears ranged from 0.0-5.0% (median:
1.0%) of all cells. Residual disease was identified and confirmed in 80/100 (80%)
patients, while the remaining 20 cases were negative for all tests. FC and IHC tests
were available in all cases and IFE in 97%. Residual disease was detected in 75/80
(94%) by IFE, in 60/80 (75%) by FC, and in 55/80 (69%) by IHC. Discordant results
between IFE and FC were identified in 15 cases: all were positive by IFE only, and
between FC and IHC in 15 cases: 10 positive by FC only and 5 positive by IHC only,
while IFE was positive in all of these cases again. In cases that were positive by IHC
only, the plasma cells were focally involving bone marrow.
Conclusions: Caution is required in interpreting a positive IFE result without evidence of residual disease by FC or IHC. In this setting, a serial IFE testing to exclude
delayed clearance of M-protein, and a radiologic study to detect active disease in
other sites would help clarify the situation. Although FC is more sensitive than IHC
(Figure 1), it still might miss focal neoplastic plasma cells, especially when they are
localized to the paracortical area (Figure 2). IFE remains the most sensitive screening test for MRD in this cohort. However, each test method had advantages and
they complemented each other. Sensitivity of FC may be increased by acquiring and
analyzing more events.
Keywords: plasma cell myeloma
Figure 2. A bone marrow biopsy from a patient with a history of plasma cell
myeloma after treatment. In this case, flow cytometry did not find any aberrant or
monotypic plasma cells, however, a focus of neoplastic plasma cells with Dutcher
bodies was identified. The cells were positive for CD138 and were kappa restricted
by ISH.
[PP-BM-04]
ACTIVATING B-RAF V600E MUTATION IN AGGRESSIVE
PEDIATRIC LANGERHANS CELL HISTIOCYTOSIS:
DEMONSTRATION BY ALLELE-SPECIFIC PCR/DIRECT
SEQUENCING AND IMMUNOHISTOCHEMISTRY
Gábor Méhes1, Gábor Irsai1, Judit Bedekovics1, Lívia Beke1,
Ferenc Fazekas1, Tímea Rózsa2, Csongor Kiss2
1
Department of Pathology, University of Debrecen Clinical Centre, Debrecen, Hungary
Department of Pediatric Hematology-Oncology, Institute of Pediatrics, University of Debrecen
Clinical Centre, Debrecen, Hungary
2
Langerhans cell histiocytosis (LCH) is a rare neoplastic disease originating from cells
characterized by antigen presenting Langerhans cell phenotype. The clinical spectrum of LCH is highly variable including localized and disseminated forms mostly
occurring in children. Recently, about 60% of LCHs were reported to carry the activating BRAF mutation V600E.
In our retrospective study we evaluated the occurrence and prognostic impact
of the V600E mutation in formaldehyde-fixed, paraffin embedded samples from
15 pediatric LCH cases treated at our institution. Allele-specific PCR and direct
sequencing was used to demonstrate the presence of V600E mutation and immunohistochemistry (IHC) using the mutant protein specific VE1 antibody clone was
performed to confirm mutant BRAF protein expression. 8/15 (53.3%) cases proved
to be BRAF mutant by any of the methods applied with a single case showing a
discrepancy (PCR negative/IHC positive). Four of the BRAF mutant cases (50.0%)
showed early dissemination and progressed to death within 43 months, while the
remaining mutant cases were stable and responded well to therapy. Wild type
BRAF cases (7/15, 46.6%) with generally comparable initial presentation were all
treated successfully.
In conclusion, activating V600E BRAF mutation can be frequently demonstrated in
pediatric LCH by both allele-specific PCR and IHC. Unfavourable risk cases potentially also responding to BRAF-inhibitory therapy can be identified by mutation testing using archival FFPE tumor samples.
Keywords: histiocytosis, molecular pathology, BRAF mutation
Figure 1. Flow cytometry identified aberrant plasma cells that were positive for
CD56 and CD117 while negative for CD19, CD45 expression. Note also the normal
plasma cells in the background. In contrast, IHC and ISH are inconclusive in this case.
72
| EAHP - 2014
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Conclusion: Our results showed that majority of CLL patients have at least one
chromosomal aberration. Hence CLL shows genetic heterogeneity, more studies
investigating the clinical importance of these abnormalities are needed to manage
the patients.
Keywords: Chronic lymphocytic leukemia, Fluorescence in situ hybridization, chromosomal aberration
[PP-BM-06]
STROMAL MACROPHAGES IN BONE MARROW
MICROENVIRONMENT IN REFRACTORY ANAEMIA AND
REACTIVE DYSERYTHROPOIESIS
Tomas Balharek, Juraj Marcinek, Peter Szepe, Lukas Plank
Department of Pathology, Jessenius Medical Faculty, Comenius University, Martin, Slovakia
Figure 1. Conventional H&E (left) and IHC staining (right) of BRAF-mutant (A)
and wild-type (B) LCH in bone marrow trephine biopsy serial sections. IHC was
performed using the V600E specific antibody clone VE1.
Refractory anaemia (RA) is a myelodysplastic syndrome (MDS) with unilinear dysplasia in erythroid lineage, leading to ineffective and insufficient erythropoiesis presenting in peripheral blood with anaemia refractory to conventional treatment. RA
is frequently considered when anaemia of unexplained cause is present in elderly
patients, representing one of the most common indications for bone marrow (BM)
trephine biopsy.
Results: The male–female ratio of 226 patients was 3.2/1. Sixty-five percent
(148/226) of patients had at least one abnormal karyotype by FISH. The following
abnormalities were noted with FISH: 13q14.3, 148/226 (65.4%); 17p13.1, 90/226
(39%); 11q22 ATM, 43/226 (19%); trisomy 12, 36/226 (15.9%); 13q34.3, 0/226
(0%). One hundred and sixty seven patients had two, 22 patients had three, 3 patients had four chromosomal abnormalities. None of the patients showed 13q34.3
aberration. 17p13.1 and 13q14.3 abnormalities were the most frequently paired
chromosomal aberrations.
Bioptic diagnostics of RA is difficult and subjective. We pointed our interest on the
BM stroma changes, because BM microenvironment play an active role in MDS
pathogenesis, supporting proliferation, survival and differentiation of transformed
haematopoietic precursors. Only limited and sometimes controversial data are
available about BM stromal changes in MDS.
Figure 2. Kaplan-Meier curves representing the overall survival of LCH (n=15)
respective the BRAF mutational status. The difference in overall survival between
wild-type and BRAF mutant cases was statistically significant (p=0.03).
[PP-BM-05]
CHRONIC LYMPHOCYTIC LEUKEMIA/SMALL LYMPHOCYTIC
LYMPHOMA: PREVALENCE OF CYTOGENETIC ABNORMALITIES
BY FLUORESCENCE IN SITU HYBRIDIZATION IN THE SOUTHERN
PART OF TURKEY
Melek Ergin1, Emine Kılıc Bagır1, Arbil Acıkalın1, Perihan Alsancak1,
Gulfiliz Gonlusen1, Semra Paydas2
1
2
Department of Pathology, Medical School, Çukurova University, Adana, Turkey
Department of Oncology, Medical School, Çukurova University, Adana, Turkey
Background and Aim: Chronic lymphocytic leukemia (CLL) is the most common
adult leukemia with survival times ranging from a few months to many decades.
Prognosis is related to clinical staging and cytogenetic findings. Conventional cytogenetic analysis of CLL reveals abnormalities in approximately one third of patients.
Fluorescence in situ hybridization (FISH) is more sensitive than conventional cytogenetics for specific chromosomal abnormalities. In present study our aim was to
evaluate the frequency and clinical significance of recurrent genetic abnormalities
in CLL patients.
Erythropoiesis is organized in erytroid island, closely related to function of macrophages within haematopoietic niches, important as for physiological as dysplastic
erythropoiesis. But macrophages represent a very heterogenous and plastic group
of cells. Many data were published about prognostic role of differentiation and
polarisation of tumour associated macrophages (TAM), including M1 - classically
activated (CD68+/HLA-DR+) and M2 - alternativelly activated (CD68+/CD163+)
TAM, in different types of heamatopoietic neoplasms and non-heamatopoietic solid
tumours.
We tried to analyse proportion and macrophage polarisation in RA type of MDS,
compared with changes present in cases showing reactive dyserythropoiesis. We
analyzed 20 cases (11 women, 9 men, age 52-87 years) of bioptically verified
RA and 20 cases of anaemia with clinically unexplained cause, showing reactive
non-dysplastic BM morphology in trephine biopsies (12 women, 8 men, age 45-85
years). Macrophages and their subtypes were detected immunohistochemically using antibodies CD68 (clone PG-M1), CD163 and HLA-DR. Their proportion was estimated semiquantitativelly and assessed histomorphometrically by point-counting
using an eyepiece with an graticule and using computerized image analysis.
All cases showed quantitative expansion of the BM stromal macrophage compartment with proportion of CD68+ macrophages ranging from 10 to 30%, but without
any statistically significant differences between studied groups. These macrophages
showed CD68+/CD163+ phenotype, usually with lower proportion of HLA-DR+ elements, but without significant differences between studied categories.
Our preliminary results show that accented proliferation and apoptosis of erythroid
precursors in both, dysplastic and reactive dyserythropoiesis, may lead to increase
of stromal macrophage quantity and these stromal changes should not be used
for discrimination of myelodysplasia and non-dysplastic erythropoiesis. Also it is
not clear, if applied symptomatic therapy of anaemia could influence macrophage
recruitment and activity in those cases. Supported by Grant VEGA 1/0209/13.
Keywords: myelodysplasia, dyserythropoiesis, macrophages
Materials-Methods: The present study included 226 CLL patients. FISH technique
was applied to bone marrow aspirations or peripheral blood materials of CLL
patients by using CLL LSI probes for del ATM (11q22.3), del 13q14.3, p 53 (del
17p13.1), trisomy 12, del 13q34.
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73
th
[PP-BM-07]
ISOLATED CONJUNCTIVAL MYELOID SARCOMA,
A CASE REPORT
1
2
Keywords: acute erythroid leukemia, Ewing sarcoma, Infant
1
Ibrahim Sari , Nurhan Sahin , Zehra Bozdag ,
Aysegul Kucukosmanoglu1
[PP-BM-09]
1
Gaziantep University, Gaziantep, Turkey
İnönü University, Malatya, Turkey
2
IMMUNOCITOCHEMICAL DIAGNOSIS OF THE SMALL B CELL
NEOPLASMS
Myeloid sarcoma is known as a tumor mass of myeloblasts or immature myeloid
cells occurring in an extramedullary site. When ophthalmic areas are involved, it is
usually located in the orbits and noted at or after the diagnosis of an underlying leukemia. We report a 54 year-old woman who had isolated right conjunctival myeloid
sarcoma without any other clinical signs and symptoms to emphasize the unusual
presentation of a conjunctival nodule of uncertain origin.
Keywords: conjunctiva, myeloid sarcoma
[PP-BM-08]
ACUTE ERYTHROID LEUKEMIA MIMICKING EWING SARCOMA/
PRIMITIVE NEUROECTODERMAL TUMOR IN AN INFANT
Razvan Lapadat1, Leslie J. Mortland2, Richard L. Tower2,
Wayne Tam3, Attilio Orazi3, Gabriela Gheorghe4
1
Department of Pathology, Medical College of Wisconsin, Milwaukee, WI
MACC Fund Center, Division of Pediatric Hematology/Oncology/Blood and Marrow Transplant,
Medical College of Wisconsin, Milwaukee, WI
3
Department of Pathology & Laboratory Medicine, Weill Cornell Medical College, New York, NY
4
Department of Pathology and Laboratory Medicine, Children’s Hospital of Wisconsin,
Milwaukee, WI
2
Background: Acute erythroid leukemia is a rare type of AML with a very aggressive clinical course. De novo cases represent only 1% of all of the AML cases. Pure
erythroid leukemia (no significant myeloblast component) is exceedingly rare in the
pediatric population. A high degree of suspicion is necessary to reach the diagnosis, particularly in small children. Ewing sarcoma (EWS)/Primitive Neuroectodermal
Tumor (PNET) is a type of small round blue cell tumor that frequently involves the
bone; it is encountered mostly in children and young adults and can mimic hematopoietic malignancies such as lymphoblastic lymphomas, diffuse large B cell
lymphomas and extramedullary AML/myeloid sarcomas.
Design: A three month old boy presented with left eye proptosis. Imaging studies
showed a prominent left orbital mass, multiple dural based enhancing lesions and a
widespread osteolytic process of the skull. In addition, the patient was found to have
multiple lytic lesions involving vertebral bodies as well as liver lesions.
Results: A biopsy of the orbital mass showed a malignant small round blue cell
tumor that was favored to represent EWS/PNET. The diagnosis was largely based on
the EM findings showing tumor cells with a glycogen distribution pattern consistent
with Ewing/PNET. FISH for t(11;22) was negative. Molecular studies (RT-PCR) failed
to detect EWSR1/FLI1 and EWSR1/ERG fusion transcripts.
Marrow showed extensive involvement by undifferentiated blast-like cells with fine
chromatin, inconspicuous nucleoli and vacuolated cytoplasm. Immunohistochemistry
showed the malignant cells to be positive for CD99 and PAS, largely positive for CD71
and focally positive for CD43 and CD45. In addition CD34, TdT,glycophorin A, hemoglobin A, megakaryocytic, monocytic and most lymphoid antigens tested were negative.
Flow cytometry showed a large CD45 negative population of cells that was positive for
CD36 and CD13. Cytogenetics showed deletion of the distal portion of chromosome 6q
and additional material attached to the long arm of chromosome 19.
In light of the molecular studies and cytogenetics (altogether not supportive of the
diagnosis of EWS/PNET) additional immunostains were performed and revealed
weak positivity for CD117, weak and focal positivity for MPO and negative glycophorin C. At this point, the diagnosis of pure erythroid leukemia/erythroblastic sarcoma
was established. Patient died 8 weeks after presentation, due to widespread metastatic disease.
Conclusion: We report an extremely rare case of pure erythroid leukemia presenting
as a multifocal myeloid/erythroblastic sarcoma, clinically and pathologically mimicking EWS/PNET family of tumors. A meticulous workup including immunohistochemical studies, flow cytometry, molecular studies and cytogenetics was necessary to reach the diagnosis. Our case illustrates that pure erythroid leukemia needs
to be considered in the differential diagnosis of small round blue cell tumors in
74
infancy. The erythroid lineage may be difficult to prove by immunophenotype since
erythroid markers can be negative in pure erythroid leukemia, due to its degree of
immaturity and lack of differentiation.
| EAHP - 2014
| 17-22 October 2014
Maja Perunicic Jovanovic1, Andrija Bogdanovic2, Ljubomir Jakovic3,
Marija Havelka4, Svetislav Tatic4, Nada Kraguljac3, Vesna Knezevic3,
Tijana Dragovic Ivancevic3, Tatjana Terzic4
1
Department of Pathology, Clinical Center of Serbia, Belgrade, Serbia
Clinic for Hematology, Clinical Center of Serbia, Medical Faculty, University of Belgrade,
Belgrade,Serbia
3
Clinic for Hematology, Clinical Center of Serbia, Belgrade, Serbia
4
Institute for Pathology, Medical faculty, University of Belgrade, Belgrade, Serbia
2
Aims: To evaluate the diagnostic value of immunocytohemistry (ICH) of the bone
marrow aspirates, flow cytometric immunophenotyping (FCIP), bone marrow histology and immunohistochemistry (IHC), in the assessment of bone marrow infiltration
in chronic lymphoid disorders and to predict the B-cell lymphoma type.
Methods: We studied the immunocytohemistry, FCIP, tissue histopathology and
IHC results from 44 patients with B-cell lymphoma seen at the Department of
Histopathology and the Clinic of Hemathology, Clinical Center of Serbia, between
January 1, 2010, and December 1, 2011, who had positive results on PB, bone
marrow, or body fluid and a corresponding diagnostic tissue biopsy specimen. A
selected panel of monoclonal antibodies was used both for immunocytohemistry,
FCIP and immunohistochemistry.
Results: The most common type of the B-cell lymphomas studied was chronic lymphocytic leukemia (CLL)/small lymphocytic lymphoma (CLL; 20) followed by nonHodgkin’s lymphoma (NHL; 22), hairy cell leukaemia (HCL; 1) and Diffuse Large B
cell Lymphoma (DLBCL; 1). Correlation between immunocytohemistry and immunohistochemistry was significant (p<0,05). There was agreement between the citomorphology, bone marrow biopsy (BMB), IHC and immunocytohemistry in 83% of
samples and in 70% when all methods were compared including FCIP. FCIP showed
the highest specificity in CLL diagnosis. IHH on BMB was the most powerful method
in diagnosis and subtipisation of small B cell neoplasms compared to FCIP and ICH.
Conclusion: ICH analysis of the BM aspirate can be used for the detection of infiltration and subtipisation of the small B cell neoplasms. FCIP is the best predictor of
involvement in CLL, whereas biopsy is superior to FCIP, ICH and bone marrow aspirates in subtipisation of the small B cell neoplasms. All methods seem to complement each other in most cases.
Keywords: non-Hodgkin lymphoma, immunocitochemistry
[PP-BM-10]
DETECTION OF POSSIBLE MULTIPLE CLONES IN PLASMA CELL
MYELOMA BY IMMUNOHISTOCHEMISTRY
Sabine Pomplun1, Ayse U. Akarca2, Linch David3, Marafioti Teresa2
1
Department of Histopathology, Kings College Hospital, London, UK
Department of Histopathology, University College Hospital, London, UK
3
2
Heterogeneity of tumour cells forming a single neoplastic cell infiltrate has been well
recognised previously and might be responsible for variations in therapy response,
relapse rate and therefore affect overall survival. It is further possible that the socalled ‘myeloma stem cells’ form part of this heterogenic group of cells. We present
a 79 year old lady with paraproteinemia and neuropathy. Bone marrow examination
showed an infiltrate of 30% lambda light chain restricted plasma cells. We were
able to demonstrate at least 3 different populations of myeloma cells by their different expression patterns by double, triple and quadruple immunostaining.
At present it is unclear whether this represents true heterogeneity or clonal evolution
as this is a semi-quantitative assessment and not a temporal one.
Keywords: Plasma cell myeloma, heterogeneity, immunohistochemistry
RELATIONSHIP OF PSGL-1 WITH PROGNOSIS IN MULTIPLE
MYELOMA PATIENTS
Figen Atalay1, Semsi Yildiz2, Elif Birtas Atesoglu3, Sema Karakus4,
Mahmut Bayık5
1
Bașkent University School of Medicine, Department of Hematology, İstanbul, Turkey
Bașkent University School of Medicine, Department of Pathology, İstanbul, Turkey
Kocaeli University School of Medicine, Department of Hematology, Kocaeli, Turkey
4
Bașkent University School of Medicine, Department of Hematology, Ankara, Turkey
5
Academic Hospital, Department of Hematology, İstanbul, Turkey
2
3
Figure 1. Quadruple staining x10 in myeloma. CD20 (blue), CD138 (red), cyclin
d1(brown) and CD3 (green) similtaneous staining in plasma cell myeloma showing
different populations with different expression profiles
[PP-BM-11]
A CLINICOPATHOLOGICAL ANALYSIS OF 8 CASES OF SPLENIC
B-CELL LYMPHOMA, UNCLASSIFIABLE (SBL-U)
Maja Perunicic Jovanovic1, Darko Antic2, Ljubomir Jakovic3,
Nada Kraguljac Kurtovic3, Tijana Dragovic Ivancevic3,
Violeta Milosevic3, Biljana Mihaljevic2
1
Department of Pathology, Clinical Center of Serbia, Belgrade, Serbia
Clinic for Hematology, Clinical Center of Serbia, Medical Faculty, University of Belgrade, Belgrade,
Serbia
3
Clinic for Hematology, Clinical Center of Serbia, Belgrade, Serbia
2
Introduction: We present a 8 cases of splenic B-cell lymphoma, unclassifiable
(SBL-U) and problems in distinction between Splenic Diffuse Red Pulp Lymphomas
(SDRPL), HCL-Variant (HCL-V) and from other splenic lymphomas.
Methods: Morphological, immunohistochemical and clinical features of all splenectomy specimens between the years 2010 and 2014 were analyzed and the lymphomas were sub-typed in accordance to 2008 WHO Classification of Hematolymphoid
Neoplasms. The medical charts and laboratory data of all SBL-U patients were reviewed. Bone marrow biopsy (BMB) histopathology, immunohistochemistry and flow
cytometry immunophenotyping available was correlated.
Results: A total of 70 cases were studied. The most common subtype of lymphoid
neoplasm was splenic marginal zone lymphoma (31 cases), followed by diffuse
large B-cell lymphoma (20 cases), splenic B-cell lymphoma, unclassifiable (8
cases), Hodgkin lymphoma (4 cases), mantle cell lymphoma (3 cases), hairy cell
leukemia (3 cases) and follicular lymphoma (1 case). All (SBL-U) patients at diagnosis had splenomegaly, lymphocytosis, thrombocytopenia, bone marrow involvement,
and were in advanced stage of disease. A predominant type of lymphoid infiltration
in bone marrow was intrasinusoidal, accompanied by interstitial or nodular infiltration in some cases.
Conclusion: Differential diagnosis of these disorders can be difficult, due to overlap
between some of these entities. Therefore, morphological and immunophenotypic
(immunohistochemistry and flow cytometry) features of BMB and splenectomy
specimen need to be correlated for a precise diagnosis and adequate therapeutic
approach.
Keywords: splenic lymphoma, unclassifiable
Introduction: Changes occur in adhesion molecules in the disease course of multiple myeloma. P-selectin glycoprotein ligand-1 (PSGL-1, CD162) works as the ligand of selectines-neutrophil adhesion molecules. The aim of this study was to
investigate the relationship between PSGL-1 expression in the bone marrow and the
known prognostic factors of multiple myeloma disease, disease stage, and survival.
This research included 63 multiple myeloma patients (females: 26, 41.3%; males:
37, 58.7%). Bone marrow biopsy samples obtained at disease diagnosis for each
patient were stained immunohistochemically in terms of CD162 expression using
standard diagnostic immunohistochemical staining methods. Laboratory results,
CD162 expression, overall survival, demographic characteristics of the disease, and
the relationship between the amount of CD162 expression and the disease stage
were evaluated.
Results: Among the 63 patients included in the study, survival rate was 82.3% for
one year; 73.2% for two years; 63.4% for 3 years; 51.7% for 4 years; 40.3% for
5 years, and 33.6% for 6 and 7 years. A statistically significant difference was not
detected between the CD162 staining ratio and disease survival (p = 0.232). A statistically significant difference was not detected between CD162 staining degree
and survival rates (p = 0.184). However, overall survival of the patients with no
CD162 expression in the bone marrow was lower than the patients whose CD162
was stained 1, 2 and 3 degrees (12.33 ± 11.49; 28.65 ± 31.44; 37.25 ± 29.32;
47.92 ± 45.29, respectively) (p < 0.001).
[PP-BM-12]
Discussion: In this study, CD162 staining and staining degree, together with the
other standard immunohistochemical stains, were shown to be beneficial in the
diagnosis of multiple myeloma disease. However, the results did not provide information about the disease course. Studies including a higher number of patients to
examine P-selectin and IL-6 levels are needed to investigate disease course.
Keywords: P-selectin ligand protein, Multiple Myeloma, Prognosis
[PP-BM-13]
BLASTIC PLASMACYTOID DENDRITIC CELL NEOPLASM: TWO
CASE
Figen Atalay1, Semsi Yildiz2, Ebru Demiralay2
1
2
Bașkent University School of Medicine, Department of Hematology, İstanbul
Bașkent University School of Medicine, Department of Pathology, İstanbul
Introduction: Blastic plasmacytoid dendritic cell neoplasm (BPDCN)was classiffied
as neoplastic disorder in the WHO 2008 (1).BPDCN originates from precursors
of plasmocytoid dentiritic cells.In pathological examination the neoplastic cells
expressed typically that CD4, CD56 and CD123 (2).BPDCN presented usually as
cutaneous and non-cutaneos involvements (3).Skin lesions may be in the forms
of maculopapullary, nodules, plaques. Non-cutaneos involvements can cause leukemic infiltration of the bone marrow or enlargement of the lypmh nodes (3,4). We
here present two patient who had both of skin and bone marrow infiltration.
Case 1: 57 years old man who was presented as multiple cutaneous lesions in his
whole body to our hematology clinic in december 2010. The lesions appears about
1 month before diagnosis. He had not other B symptoms. His laboratoary values was
normal.Bone marrow examination was revealed that only 6% neoplastic infiltaration
and lymph node enlargement was not seen in computurized tomography scaning.
Hyper-CVAD chemoterapy (CT) protocol was applied to him. His skin lesions improved after one course of CT and bone marrow infiltration was disappear after two
course of CT. He was rejected receieved further CT march 2011. In februrary 2014
he was admitted to clinic of the hematology because of severe weakness, pallor and
dsypnea.He had 80 percent of the cells was blastic cells in peripheral blood and diffuse bone marrow infiltration with neoplastic cells. After one course of Hyper-CVAD
and high dose methotrexate and cytarabine he is accepted as refractory. Unrelated
| 17-22 October 2014
| EAHP - 2014
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75
Case 2: 62 years old woman was referred as acute myelomonocytic leukemia to
our hematology clinic in june 2013. She had multiple skin lesions on her head and
extremities and hyperleukocytosis. Her bone marrow biopsy and skin biopsies was
done and diffuse neoplastic cell infiltration, which was expressed that CD4,CD56
and CD123 was seen both in the material., She was diagnosed as BPDCN. Her disease didin’t improve with four course of azacitydine, one course 2+5 CT protocole
and one course FLAG CT protocole, respectively. She was accepted as refractory
after this CT’s. She died after 2 months after the last CT. She was survial only 6
months after diagnosis.
associated clonal hematologic non-mast cell lineage disease (SM-AHNMD) showed
best responses and MCL were the worst. Therefore, in MCL patients, further evaluation of this strategy is needed. Additionally, new combination therapies including
KIT inhibitors should be improved to obtain long-term survival and better life-quality.
In conclusion, the findings of this case will be helpful for early diagnosis MCL and
intervention when the peripheral blood smears came first to the lab.
Keywords: Mast cell, Mast cell leukemia
Discussion: BPDCN is a difficult disease because of diagnosis and manegement of
treatment for clinicians. (5).The differential diagnosis includes primarily cutaneous
infiltrates composed of immature hematopoietic cells, some mature T/NK-cell lymphomas with predominant dermal involvement, Langerhans cell histiocytosis and
another myeloid neoplasm, usually chronic myelomonocytic leukemia. Best markers
for PDC identification on paraffin sections: CD68, BDCA-2, CD123, CD2AP, TCL1.
Chemotherapy protocoles of acute lymphoblastic leukemia have a little more long
survival than CT protocols for acut myeloblastic leukemia. Patients who have isolated cutaneous lesions survive longer as our first patient.
Conclusion: BPDCN is a difficult disease because of diagnosis for pathologists and
manegement of treatment for clinicians. Complete immunocytochemical staining
must be performed all patients who have cutaneous lesions with/without abnormalities in blood counts.
Keywords: Blastic plasmacytoid dendritic cell neoplasm
[PP-BM-14]
th
donor screaning is started because of he has not any sibling. We applied to him
FLAG CT protocole and he is refractory this CT.
DE-NOVO MAST CELL LEUKEMIA WITH KIT D816V
Hae In Bang, Rojin Park
Department of Laboratory Medicine, Soonchunhyang University College of Medicine, Seoul, Korea
Mast cell leukemia (MCL) is a rare and highly aggressive form of systemic mastocytosis (SM). MCL diagnosis is based on the meeting criteria for systemic mastocytosis
and presence of >=20% atypical mast cells in the bone marrow and/or >=10% in
the peripheral blood. MCL is divided into de novo and secondary forms with the
former comprised of leukemic and aleukemic variants. According to the 2013 MCL
review article, de-novo MCL are more aggressive and proliferate rapidly. Here, we
describe the clinical and laboratory features of de-novo MCL which is not an aleukemic variant. Leukemic form of MCL is the first report in Republic of Korea.
A 47-year-old male presented with fever, cough, sputum, diarrhea and abdominal
pain of one month duration. There was maculopapular rash on chest. Abdomen
computed tomography (CT) showed hepatosplenomegaly. A blood count showed a
haemoglobin of 114 g/l, leukocyte count of 3.4 x 109/l, absolute neutrophil count
(ANC) of 1.2 x 109/land platelet count of 44 x 109/l. The blood film revealed leukoerythroblastic feature with 16% atypical mast cells. Atypical mast cells display
morphologic abnormalities such as an eccentric oval nucleus, elongated cytoplasmic extensions, hypogranulated with coalescent granules, focal granule accumulations, undifferentiated immature cells with metachromatic granules and ungranulated blast. Bone marrow aspiration and trephine biopsy were performed. A bone
marrow aspirate smears showed blasts (23% of nucleated cells) and atypical mast
cells (21.4% of nucleated cells) (Fig1). The biopsied marrow was hypercellular (approximately 80%), consisting diffuse and interstitial infiltration of spindle-shaped
immature cells (Fig1). Immunohistochemically, these mast cells were positive for toluidine blue. Immunophenotyping of blasts in the bone marrow using flow cytometry
revealed positive results for CD13 (83%), CD33 (51%), CD117 (34%), CD25 (86%)
and CD203c (80%) but, negative for CD2. The patient’s karyotype was normal with
46,XY[20]. The KIT D816V mutation was identified by PCR and direct sequencing.
The diagnosis of SM can be made by satisfying one major criterion and three minor
criteria of 2008 WHO. In addition, bone marrow aspirate smears showed 20% or
more and peripheral blood smears showed 10% or more mast cells, the patient was
diagnosed with de-novo leukemic MCL. He received chemotherapies (fludarabine,
cytarabin) three times and received allogenic peripheral blood stem cell transplantation (allo-PBSCT). Remission was never achieved. He is currently being followed
up in our hospital for about 8 months and recently he was admitted for treating
neutropenic fever.
MCL express usually immature markers such as CD123, CD34, HLA-DR and display
reduced expression of CD117 and FceRI, in contrast to the more indolent forms.
KIT mutation is correlated with that immature immunophenotyping and poor prognosis. Previous study reported 23 SM patients who received allo-SCT. SM with an
76
| EAHP - 2014
| 17-22 October 2014
Figure 1. Bone marrow aspiration (Wright-giemsa stain x1000), bone marrow
biopsy (H&E stain, x200). (A) Note that the mast cells found in bone marrow
aspiration exhibited atypical morphology characterized as elongated nuclei and
hypogranular cytoplasm; (B) Hypercellular marrow with interstitial infiltration of mast
cells in a bone marrow biopsy.
[PP-BM-15]
CYTOLOGICAL AND HISTOLOGICAL BONE MARROW
EXAMINATION, A DUAL APPROACH
Veronika Kloboves Prevodnik, Mojca Velikonja, Ulrika Klopcic,
Barbara Gazic, Gorana Gasljevic
Institute of Oncology, Ljubljana, Slovenia
Background: Until 2009, at the Institute of Oncology Ljubljana, Slovenia only trephine biopsy was used as a diagnostic tool for the diagnosing of bone marrow (BM)
disorders. Since comprehensive diagnosis of BM disorders requires the integration
of various diagnostic approaches, cytological examination with flow cytometric immunophenotyping of BM aspirates was introduced into routine diagnostics according to ICSH guidelines for the standardization of BM specimens and reports.
Aims: We tried to determine the consistency of histological and cytological diagnoses
and explore the efficiency of dual examination.
Results: 985 BM aspirates and biopsies were submitted to both cytological and
histological examination. In 588 (59.7%) only non-specific, reactive changes have
been found, in 361 (36.6%) cases the lymphomatous infiltration was present, in 5
(0.5%) cases the diagnosis of chronic myeloproliferative neoplasm was suggested,
in 3 (0.3%) cases acute myeloid leukemia and in 19 (1.9%) cases dysplastic-like
changes were recognized. B cell monoclonal lymphocytic proliferations of undetermined significance were identified in 4 (0.4%) of cases, in 1 (0.1%) case metastatic
adenocarcinoma was present and in 4 (0.4%) cases the biopsy was not representative. Among cases with lymphomatous infiltration 296 (82.0%) were mature B-cell
lymphomas, 43 (11.9%) mature T-cell lymphomas, 16 (4.4%) Hodgkin lymphomas
and 6 (1.7%) precursor T-cell lymphomas.
Cytological and histological diagnoses were consistent in 713 (72.4%) cases, partly
consistent in 111 (11.3%) and inconsistent in 161 (16.3%). In 92 out of 161 (57.1%)
inconsistent cases the final diagnosis achieved at hematopathology meeting was
the same as histological diagnosis and in 69 (42.9%) cases the same as cytological
diagnosis.
Conclusions: Cytological and histological diagnoses were consistent in 72.4 % patients. In patients with partially consistent or inconsistent findings dual approach improved sensitivity and specificity of BM examination, regarding the lymphomatous
infiltration.
Keywords: bone marrow, lymphoma, diagnostics
[PP-BM-16]
LANGERHANS CELL HISTIOCYTOSIS OF TONSILLLA:
A CASE REPORT
Figen Atalay1, Eltaf Ayca Ozbal Koc2, Semsi Yildiz3
1
Bașkent University School of Medicine, Deparment of Hematology, İstanbul,Turkey
Bașkent University School of Medicine, Deparment of Otorhinolaryngology, İstanbul,Turkey
2
3
Bașkent University School of Medicine, Deparment of Pathology, İstanbul,Turkey
Introduction: Langerhans cell histiocytosis (LCH) is a rare disease and infiltration
of various organs with Langerhans cells. LCH affects usually the bone, the skin and
the ptiutary gland, occasionaly the hematopoetic system, lymph nodes, lungs. LCH
is seen mostly under 10 years of age. We here reported an adult patient who has
isolated tonsilla infiltration of LCH.
Case: 74 year old man admitted to the hematology clinic for swelling in his neck
and difficulty in swallowing. There was no pathological findigs except submandibullary lymph node enlargement about 3cm and tonsilla enlargement in physical
examination. Lymph node and excional biopsy and tonsillectomy was applied to
him. Histiocyte like cell infiltration was seen in the biopsy of tonsilla.CD3, CD20,
CD15, CD30, CD5, CD138, Lambda, Kappa, Bcl-2, ALK, CD23, CD10, Bcl-6, Keratin,
EMA, HMB-45, Cyl D1 were negative, CD68, S-100,CD1a and fascin were positive in
immunocyochemical staining. Reactive lmphocytic proliferation was seen in lymph
node biopsy. For the patients risk stratification we applied to him whole body compturize tomography (CT) scaning and boen marrow biopsy. There was no abnormal
cell infiltration in the bone marrow biopsy and lmyph node enlargement in thoracoabdominal CT scanning. His laboratory values ara mild normochromic normocyter
anemia with mild high sedimentation rate. There was no. We gave him only prednisolon 1mg/day for 6 weeks. After this treatment schedule he has no complaints
and his all laboratory valueas are normal.
Discussion: LCH is different disease and ıts diagnosis can be diffucult for pathologist. Cases of Langerhans cell histiocytosis exhibiting an immature morphology and
few associated eosinophils should also be excluded using appropriate markers
Conclusion: Isolated tonsilla involvement in adult Langerhans’ cell histiocytosis is
not reported yet. This is the first case report in the literature. There is no standard
treatment. Our patient responded well to steroid therapy
Keywords: Langerhans Cell Histiocytosis, tonsilla
[PP-BM-17]
IGG PLASMA CELL LEUKEMIA CONCOMITANT WITH
IGA HYPERGAMMAGLOBULINEMIA AND CUTANEOUS
LYMPHOPROLIFERATIVE DISORDER
Nives Jonjic1, Dora Fuckar Cupic1, Irena Seili Bekafigo2,
Ksenija Lucin1, Toni Valkovic3
1
Department of Pathology, School of Medicine, University of Rijeka, Rijeka, Croatia
Department of Cytology, Rijeka University Hospital Center, Rijeka, Croatia
3
Department of Hematology, Rijeka University Hospital Center, Rijeka, Croatia
2
Background: Primary plasma cell leukemia (PCL) is a rare and aggressive variant
of plasma cell myeloma characterized by high levels of circulating plasma cells.
Clinical presentation includes extramedullary infiltration of various tissues and
organs as a frequent complication. The disease has a fulminant course and poor
prognosis as it was the case in patient presented herein.
Patient and Methods: Peripheral blood, bone marrow and lymph node FNA and
biopsy were examined by conventional morphology and imunocytochemical and
immunohistochemical analysis at the time of hospitalization, as well as CT bone
scan, while visceral organs including skin specimen were examined post mortem.
According to the cytological and histological findings the analysis results were classified into three groups: consistent, partially consistent or inconsistent. If cytological
and histological findings coincided the diagnoses was found consistent. If cytology
and histology gave somewhat different information but still both of them favoured
either BM disease, either reactive changes the diagnoses were declared partially
consistent. In inconsistent cases the findings of one diagnostic procedure were
negative while the other suggested the presence of BM disease.
(S-100, CD1a, langerin). Notably, tumoral Langerhans cells regularly express CD4
and CD5670 and can be positive for CLA and CD123. LCH usually a systemic infiltrating disease. Isolated lymph node infiltration or hematopoietic system infiltration
is rare.
Materials and Methods: Our study included patients with BM aspirates and biopsies
obtained at the Institute of Oncology, Ljubljana Slovenia between May 2009 and
December 2013. Indications for BM sampling were mainly lymphoma staging and
prolonged, unexplained cytopenias. For each patient with BM sample cytological,
histological and final diagnoses were obtained. The final diagnosis was achieved
as consensus obtained at hematopathology meeting based on integration of clinical
data, laboratory results and the results of cytological, histological, immunophenotypic and molecular BM examination.
Results: A 66-year-old women was admitted to the hospital because of thrombocytopenia, hemopthysis, lymphadenopathy, and skin rush. Laboratory findings revealed normocytic anemia (Hb 84 g/L) and decreased platelet count
(26x109/L). Blood smear showed 26% of atypical plasma cells. Immunofixationelectrophoresis detected a monoclonal band defined as IgG-lambda light chains,
with a broad band polyclonal IgA. A bone marrow aspirate and biopsy showed an
excess of atypical plasma cells (57%) with restriction of lambda light chain. There
was no evidence of osteolytic lesions. Patient died due to splenic rupture, before
the diagnostic work-up was finished. Post-mortem examination revealed that
lymph nodes, spleen, liver, and kidney were infiltrated with atypical plasma cells
while skin biopsy revealed scattered large CD30+ lymphocytes in a background
of neutrophils and rare eosinophils. Based on all these findings, PCL (IgG-lambda
type) and lymphomatoid papulosis were diagnosed.
Conclusions: Present case is intriguing and challenging since PCL is a rare form of
myeloma. In addition in this case PCL was associated with CD30+ lymphoproliferative skin disorder and polyclonal IgA hypergammaglobulinemia.
Keywords: plasma cell leukemia, cutaneous lymphoproliferative disorder
[PP-LYMP-001]
AN IMMUNOHISTOCHEMICAL ALGORITHM RELIABLY
DISTINGUISHES BETWEEN NODAL MARGINAL ZONE
LYMPHOMA AND FOLLICULAR LYMPHOMA
Michiel Van Den Brand1, Janneke Mathijssen1, Mar Garcia Garcia2,
Konnie Hebeda1, Patricia Groenen1, Sergio Serrano2,
Han Van Krieken1
1
Department of Pathology, Radboud University Medical Center, Nijmegen, the Netherlands
Department of Pathology, Hospital del Mar-IMIM, Universitat Autònoma de Barcelona, Barcelona,
Spain
2
Differentiation of nodal marginal zone lymphoma (NMZL) from follicular lymphoma
(FL) remains a diagnostic problem in daily hematopathological practice. Multiple
new markers have become available, but they have only been tested in single
studies.
We tested the germinal center markers BCL6, CD10, LMO2, and HGAL and the marginal zone markers MNDA and IRTA-1 on a group of 50 NMZLs and 47 FLs. All FLs
| 17-22 October 2014
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77
The algorithm was subsequently validated on a second group of 13 NMZLs and 21
FLs. In this validation series, most NMZLs (85%) were classified as NMZL, whereas
most FLs (86%) were classified as FL by the algorithm. No FLs and only one NMZL
were misclassified as NMZL or FL, respectively.
Finally, the algorithm was applied to 6 cases of FL without a translocation involving
BCL2. The algorithm classified all these cases correctly as FL.
Keywords: nodal marginal zone lymphoma, follicular lymphoma, low-grade B-cell
lymphoma
th
were required to have a break involving BCL2. With the results, a diagnostic algorithm was constructed that reliably differentiated between FL and NMZL. With this
algorithm, 91% of FLs in the initial series were correctly classified as FL and only
one case was misclassified as NMZL. Of the NMZLs, 68% was correctly classified
as NMZL and only 12% was misclassified as FL. Ten cases (20%) in the NMZL group
were considered unclassifiable by the algorithm.
mutation. A common feature of these genetic changes is their ability to enhance
NF-kB activation. However, these genetic changes occur at variable frequencies in
MALT lymphoma of different anatomic sites, and the majority of MALT lymphomas
lack any of the above genetic changes. Recent somatic mutation profiling in other
B-cell lymphomas characterised by constitutive NF-kB activity, particularly activated
B-cell like diffuse large B-cell lymphoma, has identified a range of mutations in
the NF-kB pathway and other molecular processes with cooperative oncogenic
properties. In the present study, we investigated the recurrent mutations in these
molecular pathways and processes including BCR (CD79A, CD79B, CARD11), NF-kB
(TNFRSF11A, TNFAIP3, TRAF3), TLR (MYD88), plasma cell differentiation (PRDM1),
histone modifiers (CREBBP, EP300, EZH2, MLL2, MEF2B, KDM2B), antigen presentation and recognition (B2M, CD58), and TP53 in a total of 201 MALT lymphomas from
various anatomic sites. Mutation in these genes was screened by massive parallel Fluidigm Access Array PCR and Illumina MiSeq sequencing using DNA samples
from formalin-fixed paraffin-embedded lymphoma tissues. Among the 17 genes
investigated, TNFAIP3 mutation was the most common (41/201=20%), but variable in MALT lymphoma of different sites, being frequent in those from the ocular
adnexa (31/102=30%), salivary gland (6/49=12%), lung (2/10=20%) and thyroid
(2/8=25%), but absent from the stomach (0/15), skin (0/10), and small intestine
(0/7). Mutation in the remaining 16 genes was infrequent or absent in MALT lymphoma, with CARD11 in 2/201 (1%), CD58 in 2/201 (1%), CD79A in 1/201 (0.5%)
CD79B in 4/201 (2%), CREBBP in 9/201 (4%), EP300 in 4/201 (2%), EZH2 in 1/201
(0.5%), MEF2B in 3/201 (1%), MLL2 in 11/201 (5%), MYD88 in 6/201 (3%), PRDM1
in 11/201 (5%), TNFRSF11A in 6/201 (3%), TP53 in 2/201 (1%) and TRAF3 in 2/201
(1%). No mutations were observed in B2M or KDM2B. These results demonstrate a
distinct difference in the mutation profile between MALT lymphoma and other B-cell
lymphomas characterised by constitutive NF-kB activities despite their common
share of frequent TNFAIP3 mutations. The absence of the above known mutations in
a large proportion of MALT lymphoma strongly indicates that there are novel genetic
changes to be discovered in MALT lymphoma.
Keywords: MALT Lymphoma, NF-kB, Mutation
[PP-LYMP-003]
PARALLELS OF HUMAN ANGIOIMMUNOBLASTIC T-CELL
LYMPHOMAS (AITL) AND THE SJL MOUSE
Figure 1. Algorithm. Immunohistochemical algorithm for separation of nodal
marginal zone lymphoma (NMZL) from follicular lymphoma (FL). The algorithm
starts at the top with a lymphoma that is considered to be either FL or NMZL. If all
four germinal center markers (BCL6, CD10, LMO2, HGAL) are positive, a diagnosis
of FL is made. If not, IRTA-1 expression is determined. If IRTA-1 is positive, a
diagnosis of NMZL is made. If IRTA-1 is negative, MNDA and germinal center
markers are used to divide the remaining cases in three categories: NMZL for MNDA
positive cases with positivity for none or only one germinal center marker, FL for
MNDA negative cases with expression of 2 or 3 germinal center markers and lowgrade B-cell lymphoma, unclassifiable for cases that do not fit into these other two
categories.
[PP-LYMP-002]
DISTINCT MUTATION PROFILE OF MALT LYMPHOMAS
COMPARED TO OTHER B-CELL LYMPHOMAS CHARACTERISED
BY NF-B ACTIVATION
Sarah Moody1, Leire Escudero Ibarz1, Ming Wang1, Xuemin Xue1,
Naiyan Zeng1, Kim Brugger2, Yingwen Bi1, Alistair Robson3,
Shih Sung Chuang4, Sergio Cogliatti5, Hongxiang Li1, John Goodlad6,
Ming Qing Du1
1
Division of Molecular Histopathology, Department of Pathology, University of Cambridge,
Cambridge, UK
2
3
Department of Dermatopathology, St John’s Institute of Dermatology, London, UK
4
Department of Pathology, Chi-Mei Medical Centre, Tainan, Taiwan
5
Institute of Pathology, State Hospital St. Gallen, St. Gallen, Switzerland
6
Department of Pathology, Western General Hospital, NHS Lothian University Hospitals Trust,
Edinburgh, UK
MALT lymphoma is characterised by several recurrent genetic changes including t(11;18)/API2-MALT1, t(1;14)/BCL10-IGH, t(14;18)/IGH-MALT1 and TNFAIP3
78
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| 17-22 October 2014
Alina Nicolae1, Shweta Jain2, Jing Chen3, Hongsheng Wang2,
Dong Mi Shin4, Elizabeth B. Adkins6, Tomomi Sakai2,
Alexander L. Kovalchuk2, Mark Raffeld1, Jerrold M. Ward2,
Thomas A. Waldmann3, Derry Roopenian5, Elaine S. Jaffe1,
Herbert C. Morse2
1
Laboratory of Pathology, Center for Cancer Research, NCI, Bethesda, MD, USA
Virology and Cellular Immunology Section, Laboratory of Immunogenetics, NIAID, MD, USA
Lymphoid Malignancies Branch, Center for Cancer Research, NCI, Bethesda, MD, USA
4
Department of Food and Nutrition, Seoul National University, Seoul, Korea
5
Jackson Laboratory, Bar Harbor ME; Genetics Program, Tufts University Sackler School of
Graduate Biomedical Sciences, Boston, MA, USA
2
3
AITL is a TFH neoplasm commonly associated with hypergammaglobulinemia and
autoimmune manifestations. Clonal B-cell expansions occur in 10-50% of cases
and are often EBV positive. Additionally, EBV-negative clonal plasma cell proliferations have been described. Although molecularly, recurrent mutations in IDH2, TET2,
RHOA, and DNMT3A have been identified in many cases, the mechanisms that drive
and sustain the proliferation of TFH, as well as the expansion of EBV negative B-cells
are unclear. Furthermore, murine models that fully recapitulate the spectrum of human disease have not been described. SJL mice spontaneously develop lethal hematopoietic tumors, which morphologically show a confusing mixture of histiocytes,
plasma cells, centroblasts, immunoblasts and dendritic cells.
We show that TFH are expanded in the SJL mouse, along with germinal center (GC)
B cells, clonal plasma cell proliferations and serum paraproteinemia. We investigated both human AITL and murine SJL disease to analyze the importance of IL21
signaling in both entities, taking into account that TFH are a main producer of IL21.
We employed gene expression profiling (GEP) by NanoString to study IL21 and the
TFH signature in 40 cases of FFPE AITL. The GEP of 47 target genes was compared
with 10 reactive lymph nodes.
TFH and GC B-cell expansions in SJL mice were compared with C57BL/6J mice
3 months old. We used CXCR5, ICOS and PD1 as TFH markers and FAS and GL-7
as markers of GC B-cells. The dynamics of serum paraproteinemia in 60 SJL
female mice between 6 and 14 months of age were studied by serial monthly
bleeds. We compared the GEP of SJL spleens from mice 6 weeks, 6 months and
SJL mice developed expansion of TFH, B-cells, increased level of IL21 and serum
paraproteinemia. Rarely T-cell clones were also observed. The GEP identified 113
genes differentially expressed between the SJL mouse and the controls, from which
the IL21 transcript showed striking age-related upregulation. IL21 receptor-deficient mice had markedly reduced populations of CD4+ T cells and B cells, significantly lower levels of serum immunoglobulins, and reduced IL21. The proportional
representation of TFH among total CD4+ T cells was not significantly changed.
Finally, IL21R-deficient SJL mice did not develop lymphomas until they were older
than 12 months.
Rearrangements of MYC and, of high interest, BCL2 are relevant prognostic factors
in the MegaCHOEP trial. These data confirm the importance of MYC testing also in
young high-risk patients. Remarkably, breaks at BCL2 emerged as a significantly
negative prognostic indicator in younger high-risk patients in contrast to elderly
patients and elderly high-risk patients. Therefore, modifications of the spectrum of
biological risk factor testing across patient age groups are advised. The prognostic
role of BCL2, BCL6, and MYC, as well as of other biological risk factors, therefore,
must be validated in independent and biologically distinct data sets.
IL21 was the most highly expressed gene in AITL and also plays a critical role in the
development of B-cell tumors in the SJL mouse. These findings suggest that similar
mechanisms are likely to operate in AITL, especially in driving the EBV negative
B-cell proliferations. MMTV has been implicated in the SJL mouse, similar to the
frequent occurrence of EBV in AITL. SJL disease may provide a useful preclinical
model to evaluate targeted therapeutic agents that block the IL21 signaling pathway
for treatment of AITL.
Keywords: MYC, FISH, DLBCL
Keywords: Angioimmunoblastic T-cell lymphoma, Mouse model, IL21
Seung Eun Lee, So Young Kang, Young Hyeh Ko
[PP-LYMP-005]
CLONALITY ANALYSIS AND IGH MUTATIONAL STATUS IN
RECURRENT B-CELL LYMPHOMAS
Department of Pathology and Translational Genomics, Samsung Medical Center, Sungkyunkwan
University School of Medicine, Seoul, Korea
[PP-LYMP-004]
ANALYSIS OF THE PROGNOSTIC RELEVANCE OF MYC,
BCL2 AND BCL6 GENE REARRANGEMENTS AND PROTEIN
EXPRESSION IN YOUNG HIGH-RISK DLBCL PATIENTS IN THE
MEGACHOEP TRIAL OF THE DSHNHL
Heike Horn1, Marita Ziepert2, Annette M. Staiger1,
Martin Wartenberg3, Peter Möller4, Alfred C. Feller5,
Wolfram Klapper6, Michael Hummel7, Harald Stein8,
Martin Leo Hansmann9, Markus Loeffler2, Norbert Schmitz10,
German Ott1, Andreas Rosenwald3
1
Institute for Medical Informatics, Statistics, and Epidemiology, Universität Leipzig, Leipzig, Germany
3
Institute of Pathology, Universität Würzburg, Würzburg, Germany
4
Institute of Pathology, Universitätsklinikum Ulm, Ulm, Germany
5
Institute of Pathology, Universitätsklinikum Schleswig-Holstein, Campus Lübeck, Lübeck, Germany
6
Institute of Pathology, Hematopathology Section and Lymph Node Registry, Universitätsklinikum
Schleswig-Holstein, Campus Kiel, Kiel, Germany
7
Institute of Pathology, Campus Benjamin Franklin, Charité Universitätsmedizin, Berlin, Germany
8
Pathodiagnostik, Berlin, Germany
9
Institute of Pathology, Universitätsklinikum Frankfurt, Frankfurt, Germany
10
Department of Hematology, Asklepios Klinik St. Georg, Hamburg, Germany
2
Rearrangements of MYC occur in 5-10% of diffuse large B-cell lymphomas (DLBCL)
and – in contrast to BCL2 and BCL6 translocations - confer an increased clinical risk
to CHOP and R-CHOP treated patients. In addition, simultaneous protein overexpression of MYC and BCL2 has more recently been identified as a robust and reproducible marker of increased clinical risk across studies. The larger part of these studies,
however, have enrolled only small patient cohorts and/or, of importance, only older
patients. The prognostic impact of MYC, BCL2 and BCL6 translocations and protein
expression was, therefore, assessed in a clinical trial of younger high-risk patients
with DLBCL for the first time.
Tumor samples from 112 patients with de novo DLBCL enrolled in the MegaCHOEP
trial (young high-risk patients 18-60 years, aaIPI 2 or 3; Schmitz et al. 2012, Lancet
Oncol) of the DSHNHL were immunostained with antibodies directed against
MYC, BCL2, and BCL6, and hybridized by FISH with DNA probes suitable to detect
MYC-, BCL2- and BCL6- breaks. The MegaCHOEP study randomized patients to
8xCHOEP-14 or sequential high-dose therapy supported by repeated infusions of
autologous stem cells. The median patient age was 48 years, and 27% of patients
scored an aaIPI of 3.
GEP was successful in 39/40 cases. High expression of IL21 was observed in all
cases of AITL but one. Furthermore, overexpression of IL21R, BATF, IL4 and ICOS,
genes that promote the expression of IL21 by TFH, was identified. A significant number of cases also exhibited increased expression of STAT5 and JAK3, both of which
are activated downstream of the IL21R.
10/14 (71%) MYC rearranged DLBCL. Presence of BCL2 breaks (RR=4.7, 95%
CI: 1.8-12.2) and MYC breaks (RR=2.4, 95% CI: 0.8-7.5) but not of BCL6 breaks,
were associated with inferior overall survival in univariate and multivariate analyses adjusted for aaIPI and treatment arm. Protein overexpression of MYC >=30%
(RR=2.4, 95% CI: 0.9-6.5), but not of BCL2 (>=60%) or BCL6 (>=30%) indicated inferior overall survival. BCL2 overexpression was associated with inferior
EFS (RR=2.2, 95% CI: 0.9-5.5) and PFS (RR=2.8, 95% CI: 1.0-8.2). If the same
cutpoint for BCL2 used for a similar analysis in elderly patients treated on the
RICOVER-60 trial (Horn et al. Blood 2013) was applied, no differences in EFS or
PFS were observed.
12 months of age with those of spleens from young, healthy NFS.V+ mice, an
inbred subline of NIH Swiss outbred mice. To further prove that IL21 produced by
TFH plays a central role in the pathogenesis of SJL disease, a knockout (IL21r-/-)
was generated.
Background: Distinction between recurrent B-cell lymphoma and second primary
lymphoma has clinical importance because the management of both entities was
different. However, it may not be possible to distinguish both components by histological characteristics alone. Immunoglobulin (Ig) gene rearrangements remain
largely unmodified during clonal proliferation of neoplastic B-cells irrespective of
cellular phenotype. On the basis of this phenomenon, the analysis of Ig gene rearrangements help to determine whether or not both lymphoma components share a
common clonal origin.
Materials and Methods: The clonal relationship of the both lymphoma components
was analyzed using BIOMED-2 multiplex polymerase chain reaction (PCR) assays
in formalin fixed, paraffin embedded tissues of 30 patients with recurrent B-cell
lymphoma at the single institute from January 1995 through December 2013.
Furthermore, sequencing of the amplified immunoglobulin heavy chain (IgH) gene
products was performed in cases which difference of monoclonal PCR products size
was less than 5bp. Because:
Results: In 30 cases of primary B-cell lymphoma, monoclonal rearrangements of Ig
genes were detectable in all 30 cases (100%) and in the corresponding recurrent
B-cell lymphoma, clonal Ig rearrangements were detectable in 28 cases (93.3%) using complete set of BIOMED-2 reactions (3 IgH, 1IGK and 1IGL). After the PCR-based
Genescan assay, the exact size of the CDR1 or CDR2 or CDR3 fragments of Ig genes
were compared with the both components in each case. The same size was observed in 18/30 cases (60.0%), whereas size was different in 12/30 cases (40.0%).
Out of the 12 cases showing different PCR product size, nucleotide sequencing of
IgH gene was performed in 7 cases. Nucleotide sequencing of the amplified IgH
gene CDR1, CDR2, and CDR3 region products was carried out in 1, 3, and 5 cases,
respectively. In case 3, sequencing of all regions was performed. The same VDJ segments were observed in 3 out of 7 cases. Out of 3 clonally related cases, one case
carried unmutated. Out of 4 clonally unrelated cases, 3 cases carried unmutated.
Finally, by both comparison of Ig gene fragment length and IgH gene sequencing
analysis, 21/30 cases (70.0%) clonally related, whereas 9 cases (30.0%) were clonally unrelated irrespective of phenotypic change.
Conclusion: To our knowledge, this is the largest study to date assessing the clonal
relationship of recurrent B-cell lymphoma. Analyzing IgH rearrangements seems
to be the most accurate way to determine the clonal relationship of a recurrent
lymphoma.
Keywords: Recurrent B-cell lymphoma; Clonality; Ig gene rearragements
Rearrangements of MYC, BCL2 and BCL6 were detected in 13.6%, 20.7% and
30.9% of DLBCL, respectively. A double or triple hit constellation occurred in
| 17-22 October 2014
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79
Primary/
Secondary
site
Primary/Secondary
diagnosis
Time to
relapse
(month)
Death
Clonality
Duodenum/
Oral cavity
MZBCL/DLBCL
9
No
Unrelated
No.
Age at
diagnosis
3
54
4
63
LN/LN
SLL/DLBCL
100
Yes
Unrelated
26
71
Brain/BM
DLBCL/
Lymphoplasmacytic
31
No
Related
Table 2. Results of clonal identity analysis according to histologic type in 27
cases showing same phenotype
Primary/Secondary diagnosis
DLBCL/DLBCL
Clonality
No. of case
Related
6
Unrelated
5
FL/FL
Related
4
(n=4)
Unrelated
0
(n=11)
MCL/MCL
(n=8)
MZBCL/MZBCL
(n=3)
SLL/SLL
(n=1)
Related
7
Unrelated
1
Related
2
Unrelated
1
Related
1
Unrelated
0
independent of the IPI. Among 31 TNF--positive DLBCL cases, 27 (87%) were
positive and 4 (13%) were negative for TNFR1. Both TNF-- and TNFR1-positive
cases were significantly correlated with a poorer OS than the TNF--positive but
TNFR1-negative cases (P = 0.0191, log-rank test). Twenty-seven cases (45%)
with both TNF-- and TNFR1-positive subtype of DLBCL, NOS had a poorer prognosis for OS (P < 0.0001, log-rank test) and PFS (P = 0.0050, log-rank test) than
the 33 cases (55%) with the remaining subtypes (Figure 2C), and both TNF-- and
TNFR1-positive subtype of DLBCL, NOS was also shown to be a significant prognostic factor for OS (P < 0.0001) and PFS (P = 0.0060) independent of the IPI.
Conclusion: TNF--positive DLBCL, NOS constitute a unique, clinically aggressive
type of DLBCL, NOS. In addition, both TNF-- and TNFR1-positive subtype of DLBCL,
NOS was correlated with poorer prognosis. In addition to the IPI, the prognosis of
patients can be more accurately identified by evaluating both TNF- and TNFR1
expression. TNF--targeting biological agents may be somewhat effective for TNF-positive patients.
Keywords: diffuse large B-cell lymphoma not otherwised specified, TNF-, TNF-
receptor
th
Table 1. Results of clonal identity analysis in 3 cases of different histologic
type of recurrent B-cell lymphoma
[PP-LYMP-006]
TNF- AND TNF- RECEPTOR 1 EXPRESSION PREDICTS POOR
PROGNOSIS OF DIFFUSE LARGE B-CELL LYMPHOMA, NOT
OTHERWISE SPECIFIED
Shoko Nakayama, Taiji Yokote, Motomu Tsuji, Toshikazu Akioka,
Takuji Miyoshi, Yuji Hirata, Nobuya Hiraoka, Kazuki Iwaki,
Ayami Takayama, Uta Nishiwaki, Yuki Masuda, Toshiaki Hanafusa
Figure 1. Immunohistochemistry for tumor necrosis factor (TNF-α), TNF-α receptor
1 (TNFR1), and TNF-α receptor 2 (TNFR2) expression in DLBCL, NOS (original
objective magnification, 40×). (A) Representative TNF-α-positive DLBCL, NOS; (B)
Representative TNF-α-negative DLBCL, NOS; (C) Representative TNFR1-positive
DLBCL, NOS; (D) Representative TNFR1-negative DLBCL, NOS; (E) Representative
TNFR2-positive DLBCL, NOS; (F) Representative TNFR2-negative DLBCL, NOS.
Osaka Medical College, Osaka, Japan
Background: Tumor necrosis factor- (TNF-) is a key cytokine involved in inflammation, immunity, cellular homeostasis, and tumor progression. Circulating
tumor necrosis factor- (TNF-), TNF- receptor 1 (TNFR1) and TNF- receptor
2 (TNFR2) have recently been reported to be a significant predictor of treatment
outcomes in diffuse large B-cell lymphoma, not otherwise specified (DLBCL,
NOS) patients, respectively. However, until now, there have been no reports regarding the expression of TNF- and TNFRs in DLBCL, NOS cells in situ. (Aims)
To examine the expression of TNF-, TNFR1 and TNFR2 in DLBCL, NOS cells and
analyze the relationship between the expression of TNF- and TNFRs and the
prognosis of the patients.
Methods: Sixty lymphoma tissue specimens from patients with DLBCL, NOS were
immunostained with antibodies against TNF-, TNFR1, and TNFR2. The relationship
between the expression of TNF- and TNFRs and the prognosis of the patients were
analyzed using the Kaplan–Meier method the Cox proportional hazards regression
model.
Results: Among the 60 cases, 38 (63%) were immunohistochemically positive
and 22 (37%) were negative for TNF-. Further, 31 cases (52%) were positive
and 29 (48%) were negative for TNFR1, and 49 (82%) were positive and 11 (18%)
were negative for TNFR2 (Figure 1). The TNF--positive cases had poorer OS
(P = 0.0019, log-rank test) and PFS (P = 0.0078, log-rank test) than the TNF-negative cases (Figure 2A). The TNFR1-positive cases were significantly correlated with a poorer OS (P = 0.0006, log-rank test) than the TNFR1-negative
cases. The TNFR1-positive patients tended to have a poorer PFS than the TNFR1negative patients, although the difference between the 2 groups was not significant (Figure 2B). The TNFR2-positive cases tended to have a poorer OS than the
TNFR2-negative cases, although the difference was not significant. No difference
in PFS was observed between the 2 groups. TNF- expression in tumor cells was
a significant prognostic factor for OS (P = 0.0006) and PFS (P = 0.0053) and
was independent of the International Prognostic Index (IPI). TNFR1 expression in
tumor cells was also a significant prognostic factor for OS (P = 0.0005) and was
80
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Keywords: lymphoid enhancer-binding factor-1 (LEF-1), SOX11, immunoglobulin
superfamily receptor translocation associated-1 (IRTA-1)
Table 1. Positive immunohistochemical staining of three antibodies in small
B-cell lymphomas
LEF-1
SOX11
IRTA-1
24/26
SLL
22/26
0/26
MCL
0/12
11/12
7/12
MZL
1/28
0/28
28/28
FL
0/24
0/24
8/24
LPL
0/2
0/2
0/2
HCL
0/13
0/13
0/13
[PP-LYMP-008]
Figure 2. (A) Prognosis of DLBCL, NOS patients, according to their TNF-α
expression. (B) Prognosis of DLBCL, NOS patients, according to their TNFR1
expression. (C) Prognosis of TNF-α-positive DLBCL, NOS patients, according to
their TNF-α and TNFR1 expressions.
MOLECULAR CHARACTERIZATION OF DIFFUSE LARGE B-CELL
LYMPHOMA RECURRENCES: CLONAL RELATIONSHIP AND
DIFFERENT MODES OF TUMOR EVOLUTION
Conclusions: SOX11 nuclear expression is a specific marker for diagnosis of MCL
including the cases with unusual immunophenotype or gene translocation. LEF-1
nuclear expression can differentiate SLL from other small B-cell lymphomas especially from MCL. IRTA-1 cytoplasmic and membranous staining is helpful to differentiate MZL from LPL and HCL; IRTA-1 also shows positivity in some cases of SLL,
MCL and FL. These three now immunohistochemical antibodies increase our ability
to diagnosis small B-cell lymphoma.
Darius Juskevicius, Joel Gsponer, Alexander Rufle,
Stephan Dirnhofer, Alexandar Tzankov
Institute of Pathology, University Hospital Basel, Basel, Switzerland
[PP-LYMP-007]
EXPRESSION OF THREE NEW IMMUNOHISTOCHEMICAL
MARKERS IN DIFFERENT SMALL B-CELL LYMPHOMAS
Xiaohong Mary Zhang
Geisinger Medical Laboratories, Geisinger Health System, Department of Pathology, Bloomsburg,
PA 17815, USA
Background: Small B-cell lymphoma is a heterogeneous group of lymphoproliferative malignancies which have different clinical behaviors and treatments. It is important to differentiate individual B-cell lymphoma in order to apply the best treatment
and management. Morphology and immunohistochemistry are the primary tools
used for diagnosing lymphoma. There is a characteristic pattern of expression with
immunohistochemical antibodies in most well defined small B-cell lymphomas, but
new and sometimes more specific antibodies are developed, which enhance the
probability for diagnosis or can act as an alternate marker in unusual cases in which
a small B-cell lymphoma does not present with characteristic immunohistochemical
staining.
Introduction: Diffuse large B-cell lymphoma (DLBCL) is the most common lymphoma in adults. It is a heterogenous disease with diverse clinical courses, moprhologic
appearances, pheno- and genotypes. More than half of the patients can be cured
with standard therapeutic regimens. However, a substantial number of patients experience a recurrence. In general, lymphoma recurrences are considered to represent a relapse of the original neoplasm, but this concept has recently been challenged by demonstration of clonally unrelated relapses by immunoglobulin (heavy
chain) gene (IGH) rearrangement analysis. Although being the most widely used
method for clonality testing, IGH fragment length analysis has a very limited output
of information not giving any insight about further genetic tumour heterogeneity and
molecular evolution in time. Moreover, the result is based only on a single gene in
the whole genome and, therefore, it is error-prone.
We applied a genome-wide copy number aberration testing and targeted deep sequencing on 16 paired primary and relapsed DLBCL aiming to provide an unambigous answer about the existence of clonally unrelated recurrences and molecular
changes at relapse. Additionally, we profiled 10 cases of non-relapsing DLBCL and
compared their molecular profiles to the former searching for genetic markers at
diagnosis, which could predict lymhoma relapse.
Design and Methods: The expression of three new antibodies, lymphoid enhancerbinding factor-1 (LEF-1), SOX11 and immunoglobulin superfamily receptor translocation associated-1 (IRTA-1), were examined in 105 cases of different small B-cell
lymphomas including 26 cases of small lymphocytic lymphoma (SLL), 12 cases of
mantle cell lymphoma (MCL), 28 cases of marginal zone lymphoma (MZL), 24 cases
of follicular lymphoma (FL), 2 cases of lymphoplasmacytic lymphoma (LPL) and 13
cases of hairy cell leukemia (HCL) in the bone marrow biopsy.
Methods: Genomic DNA extracted from formalin-fixed paraffin-embedded tissue
with at least 70% tumor content was used for the analysis. Array-comparative genomic hybridization was utilized to detect chromosomal copy number aberrations
and to determine clonal relationship status of each primary-relapse pair. This result
was then verified by IGH gene fragment length analysis. Directed deep-sequencing
(IonTorrent) was performed on a custom hot-spot panel consisting of the most frequently mutated genes in DLBCL.
Results: Nuclear expression of LEF-1 was detected in 85% of SLL cases (22/26),
4% of MZL cases (1/28) and none of the cases of MCL, FL, LPL and HCL. Nuclear
staining of SOX11 was found in 92% of MCL cases (11/12) including a case of
negative t(11;14) by FISH study. The case of MCL with negative SOX11 nuclear
staining had positive t(11;14) by FISH and positive for BCL-1 by immunohistochemical staining but negative for CD5 by flow cytometry study and immunohistochemical
staining. SOX11 was negative in all cases of SLL including one case with weakly
positive BCL-1 immunochemical stain, MZL, FL, LPL, and HCL. Cytoplasmic and
membranous expression of IRTA-1 was detected in all cases of MZL (28/28), and
also in 92% of SLL cases (24/26), 58% of MCL cases (7/12) and 33% of FL cases
(8/24), although most of positive stain was weak in cases of MCL and FL. IRTA-1
was negative in cases of LPL (0/2) and HCL (0/13).
Results: Among 16 cases of recurring DLBCL, 2 relapses (13%) were clonally unrelated to the primary disease by copy number aberration or/and IGH fragment lenght
analysis. Different modes of tumor progression could be detected within the clonally related set of relapses, providing evidence of a divergent evolution from the
common early progenitor cell population in 4/16 (25%) and linear progression in
10/16 (62%) of the cases. Also, in our study gain of the short arm of chromosome
7 was significantly associated with DLBCL relapse. This chromosomal region contains the genes CARD11, PDGF and IL6 - all previously associated with lymphomas.
Moreover, our cohort of non-relapsing DLBCL showed more complex karyotypes
and signicantly more cytogenetic abnormalities compared to genomes of relasping
DLBCL. Thus, comparison of copy number aberration and point mutation profiles
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between relapsing and non-relapsing tumors yielded significant differences, which
could be useful for understanding the genetic basis of DLBCL recurrence.
[PP-LYMP-010]
Conclusions: We provide clear evidence for the existence of clonally unrelated relapses in DLBCL. These findings might be of clinical importance since relapses are
usually treated more aggressively than primary neoplasms with a significant treatment-related morbidity. Moreover, the high definition clonal analysis provides new
insights into the molecular profiles of tumors in vivo and, therefore, can contribute
to the development of more personalized approaches for cancer prognostication
and treatment.
LYMPHOID ENHANCER-BINDING FACTOR 1 (LEF1) IN
DIAGNOSIS OF CHRONIC LYMPHOCYTIC LEUKEMIA/SMALL
LYMPHOCYTIC LYMPHOMA
Catalina Amador Ortiz, Kristy L. Wolniak, Loann C. Peterson,
Charles L. Goolsby, Juehua Gao, Yi Hua Chen
Department of Pathology, Northwestern University Feinberg School of Medicine, Chicago, IL, USA
Keywords: lymphoma, DLBCL, clonal relationship
[PP-LYMP-009]
HIGH PREVALENCE OF ONCOGENIC MYD88 AND CD79B
MUTATIONS IN DIFFUSE LARGE B-CELL LYMPHOMAS
PRESENTING AT IMMUNE-PRIVILEGED SITES
Willem Kraan1, Hugo Horlings1, Martine Van Keimpema1,
Esther Schilder Tol1, Monique Oud1, Arnold Noorduyn2,
Cornelis Scheepstra3, Philip Kluin4, Marie Jose Kersten1,
Marcel Spaargaren1, Steven Pals1
1
Department of Pathology, Academic Medical Center, University of Amsterdam, Amsterdam,
The Netherlands
2
Department of Pathology, Dordrecht, The Netherlands
3
Department of Pathology, OLVG, Amsterdam, The Netherlands
4
Department of Pathology, UMCG, Groningen, The Netherlands
Activating mutations in CD79 and MYD88 have recently been found in a subset of
diffuse large B-cell lymphoma (DLBCL), identifying B-cell receptor and MYD88 signalling as potential therapeutic targets for personalized treatment. Here, we report
the prevalence of CD79B and MYD88 mutations and their relation to established
clinical, phenotypic and molecular parameters in a large panel of DLBCLs. We show
that these mutations often coexist and demonstrate that their presence is almost
mutually exclusive with translocations of BCL2, BCL6 and cMYC, or Epstein-Bar virus infection. Intriguingly, MYD88 mutations were by far most prevalent in immuneprivileged site-associated DLBCL (IP-DLBCL), presenting in central nervous system
(75%) or testis (71%) and relatively uncommon in nodal (17%) and gastrointestinal
tract lymphomas (11%). Our results suggest that MYD88 and CD79B mutations are
important drivers of IP-DLBCLs and endow lymphoma-initiating cells with tissuespecific homing properties or a growth advantage in these barrier-protected tissues.
Keywords: DLBCL, testis, CNS
Objective: The diagnosis of chronic lymphocytic leukemia and small lymphocytic
lymphoma (CLL/SLL) can be established in most cases based on the morphologic
and immunophenotypic characteristics. However, the differential diagnosis of CLL/
SLL from its mimickers can be challenging in some cases due to overlapping morphologic and immunophenotypic features. Lymphoid-enhancer-binding factor 1
(LEF1) is a nuclear protein expressed in mature T cells and pro-B cells but not
mature B cells, which by gene expression profiling was found to be overexpressed
in cases of CLL/SLL. We evaluated the utility of LEF1 in the diagnosis of CLL/SLL, by
(1) assessing LEF1 by immunohistochemistry (IHC) in a series of B-cell lymphomas
and (2) developing a flow cytometry assay for the detection of LEF1.
Methods: Immunohistochemical staining of LEF1 was performed on paraffin-embedded bone marrow (BM) and lymph node (LN) sections of 310 B cell lymphomas, including 102 CLL/SLL cases and 208 other B-cell lymphomas (a subset of the IHC data was
previously published in: Modern Pathology. 2011 Nov;24(11)). Antigen retrieval and
immunohistochemical staining were performed on an automated immunostainer, using monoclonal anti-LEF1 (clone: EPR2029Y; Epitomics, Burlingame, CA, USA).
Subsequently, flow cytometric analysis of LEF1 was performed in 64 patient samples (18 BM, 22 LN and 24 peripheral blood samples) by two independent reviewers blinded to the morphologic findings. A qualitative and quantitative analysis was
performed by comparing the staining intensity and the ratios of the median fluorescence intensities (MFI) of LEF1 in B cells of interest to internal reference cell
populations. The results were correlated with the pathologic diagnosis.
Results: By IHC, nuclear staining of LEF1 was observed in all 102 CLL/SLL cases
(Figure 1). LEF1 also highlighted the morphologically inconspicuous SLL component
in three composite lymphomas and was positive in two CD5- SLL cases and a CD5-/
CD20- SLL case. LEF-1 was found to be negative in all other low-grade lymphomas,
including 56 mantle cell lymphomas (MCL), 36 marginal zone lymphomas (MZL)
and 33 low-grade follicular lymphomas (FL). A subset of high grade FL (6 of 12) and
diffuse large cell lymphomas (27 of 71) showed variable staining for LEF1.
The flow cytometry assay for LEF1 effectively distinguished positive LEF1 staining
in T cells from negative staining in NK and B cells in normal PB samples (Figure 2).
In patient samples, all 25 cases of CLL/SLL and all 5 monoclonal B lymphocytosis
of CLL phenotype were LEF1 positive by flow cytometry (Figure 3). No LEF1-positive
B cells were identified in the remaining cases including 21 cases of other types of
small B cell lymphomas (6 MCL, 4 FL, 4 MZL, 3 lymphoplasmacytic lymphoma and
4 unclassifiable low-grade B-cell lymphomas) and 13 cases with no evidence of
lymphoma. Quantitative analysis using a B to NK MFI ratio of 1.50 and B to T MFI
ratio of 0.45 separated CLL/SLL cases from non-CLL lymphomas.
Conclusions: LEF1 is a highly sensitive and specific marker for CLL/SLL among
small B-cell lymphomas. Both IHC and flow cytometric analysis of LEF1 distinguish
CLL/SLL from other small B-cell lymphomas, and may serve as useful tools in the
differential diagnosis of small B-cell lymphomas.
Keywords: chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL),
flow cytometry, LEF1
Figure 1. MYD88 and CD79B by tumour localization. Prevalence of mutations
in MYD88 and CD79B by tumour localization. Percentage of ABC DLBCL with
MYD88 and/or CD79B mutations at different anatomical sites. Prevalence of MYD88
mutations in central nervous system (CNS) and testis was significantly different from
that in the lymph node and gastrointestinal (GI) tract (***P<0.001 by Fisher’s exact
test).
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Figure 1. LEF1 immunohistochemistry (IHC) in chronic lymphocytic leukemia/
small lymphocytic lymphoma (CLL/SLL) and mantle cell lymphoma (MCL). Upper
panel: A CLL/SLL case shows uniform strong nuclear staining for LEF1 in nearly
100% of neoplastic cells (right). Lower panel: A MCL case is negative for LEF1 with
only rare admixed LEF1+ T cells (right).
TARGETING B CELL RECEPTOR- AND CHEMOKINECONTROLLED ADHESION AND MIGRATION IN LYMPHOMA
Martin F. De Rooij1, Annemieke Kuil1, Steven P. Treon2,
Marcel Spaargaren1, Steven T. Pals1
1
Department of Pathology, Academic Medical Center, University of Amsterdam and Lymphoma and
Myeloma Center Amsterdam-LYMMCARE, Amsterdam, the Netherlands
2
Bing Center for Waldestrom’s Macroglobulinemia, Dana Farber Cancer Institute, Harvard Medical
School, Boston, MA, USA
Figure 2. Flow cytometric analysis of LEF1 in a normal peripheral blood sample.
A: CD19 vs. CD5 dot plot shows the gating of B cells (bright CD45+/low side scatter/
CD19+/CD5- or bright CD45+/low side scatter/CD19+/CD5+), T cells (bright CD45+/
low side scatter/CD19-/CD 5+), and NK cells (bright CD45+/low side scatter/CD19-/
CD 5-). B: B cells show negative LEF1 staining as compared to the T cells. A very
small subpopulation of T cells shows negative LEF1 staining which overlaps with B
cells (arrow). C: NK cells show negative LEF1 staining as compared to the T cells.
D: NK cells show negative staining for LEF1, which overlaps with B cells.
The pathogenesis of most types of B-cell malignancies is dependent on signaling by
the B cell antigen receptor (BCR) and/or other growth and survival signals provided by
the tumor microenvironment. In patients suffering from various B-cell malignancies,
in particular chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), and
Waldenström’s macroglobulinemia (WM), high objective response rates were recently
obtained in clinical trials with the BCR signalosome inhibitors Ibrutinib (Bruton’s tyrosine kinase-BTK-inhibitor) and Idelalisib (PI3K inhibitor), which both result in a rapid
and sustained reduction in lymphadenopathy accompanied by transient lymphocytosis. We have previously demonstrated that BTK plays a key role in regulating BCRand chemokine-controlled adhesion and migration in B cells by controlling integrin
activity. We now investigated the effects of BTK and PI3K inhibition in CLL, MCL, and
WM. We show that Idelalisib, but not Ibrutinib, strongly reduces proliferation while no
direct cytotoxicity was not observed at clinically achievable plasma concentrations.
Furthermore, we demonstrate that both drugs inhibit BCR-controlled signaling and
integrin 41-mediated adhesion to fibronectin and VCAM-1, and that Ibrutinib, but
not Idelalisib, also inhibits CXCL12-controlled integrin-mediated adhesion and migration. Taken together, our data indicate that inhibition of BTK and PI3K overcomes
BCR-controlled integrin-mediated retention of malignant B cells in their growth- and
survival-supporting bone marrow microenvironment, which results in lymphoma regression. These findings indicate disrupted homing to and retention within lymphoid
organs as a key mechanism of action of these novel drugs.
[PP-LYMP-011]
Keywords: adhesion, migration, integrin
Figure 1. BTKi. Inhibition of BTK (or PI3Kδ) impairs BCR-controlled integrinmediated adhesion and chemokine (CXCL12, CXCL13, and CCL19)–induced
adhesion and migration of lymphoma cells. Consequently, BTKi overcomes BCRand chemokine-controlled integrin-mediated retention in their growth- and survivalsupporting LN and BM microenvironment, which results in their egress from these
protective niches into the circulation (peripheral blood), and will prevent chemokinedriven homing into these niches, resulting in lymphoma regression.
[PP-LYMP-012]
ANALYSIS OF MICROSATELLITE INSTABILITY IN GASTRIC MALT
LYMPHOMA
Figure 3. Flow cytometric analysis of LEF1 in a chronic lymphocytic leukemia/
small lymphocytic lymphoma (CLL/SLL). A: CD19 vs. CD5 dot plot shows a CD5+
and a CD5- B cell population, as well as a T cell and a NK cell population. B: CD5+
B cells show positive LEF1 staining as compared to the NK cells. C. CD5+ B cells
show positive LEF1 staining as compared to the internal CD5- B cells. D. T cells in
this case show a distinct bimodal staining for LEF1. The LEF1 staining in CD5+ B
cells overlaps with the LEF1+ T-cell peak. E: Bone marrow aspirate showing sheets of
small lymphocytes with condensed chromatin and scant cytoplasm. F. Bone marrow
core biopsy showing abnormal lymphoid infiltrate with a proliferation center. The
lymphocytes are positive for LEF1 by immunohistochemical stain (inset).
Annemarie Degroote1, Knippenberg Lies1, Vanderborght Sara2,
Spaepen Marijke4, Matthijs Gert4, Schaeffer David5, Owen David5,
Libbrecht Louis3, Kathleen Lambein3, De Hertogh Gert1,
Tousseyn Thomas2, Sagaert Xavier2
1
KU Leuven, Dept of Imaging and Pathology, Translational Cell and Tissue Research, Leuven,
Belgium
2
UZ Leuven, Dept of Pathology, Leuven, Belgium
3
UGent, Dept of Pathology, Ghent, Belgium
4
UZ Leuven, Dept of Human Genetics, Leuven, Belgium
5
University of British Columbia, Dept of Pathology, Canada
In Helicobacter pylori gastritis, continuous antigenic stimulation triggers a sustained
B-cell proliferation. Errors made during this continuous DNA replication are generally corrected by the DNA mismatch repair mechanism. Failure of this mismatch
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repair mechanism has been described in hereditary non-polyposis colorectal cancer (HNPCC) and results in a replication error phenotype. Inherent to their instability during replication, microsatellites are the best markers of this replication error
phenotype. We aimed to evaluate the role of defects in the DNA mismatch repair
(MMR) mechanism and microsatellite instability (MSI) in relation to the most frequent genetic anomaly, translocation t(11;18)(q21;q21), in gastric mucosa-associated lymphoid tissue (MALT) lymphoma. Therefore, we examined 10 microsatellite
loci (BAT25, BAT26, D5S346, D17S250, D2S123, TGFB, BAT40, D18S58, D17S787
and D18S69) for instability in 28 patients with MALT lymphomas. In addition, these
tumors were also immunostained for MLH1, MSH2, MSH6 and PMS2, as well as
screened for the presence of t(11;18)(q21;q21) by real-time polymerase chain reaction (RT-PCR).
We found MSI in 5/28 (18%) lymphomas, with MSI occurring in both t(11;18)
(q21;q21)-positive and -negative tumors. One tumor displayed high levels of instability, and, remarkably, this was the only case displaying features of a diffuse large B-cell
lymphoma. All microsatellite unstable lymphomas showed a loss of MSH6 expression.
In conclusion, our data suggest that a MMR-defect may be involved in the development of gastric MALT lymphomas, and that a defect of MSH6 might be associated
with those MSI-driven gastric lymphomas.
Keywords: gastric MALT lymphoma, microsatellite instability
[PP-LYMP-013]
BCL2 TRANSLOCATION IN PRIMARY CUTANEOUS FOLLICULAR
CELL LYMPHOMA (PCFCL) AND ITS PROGNOSTIC
SIGNIFICANCE: A MULTICENTER STUDY ON BEHALF OF
GRUPPO ITALIANO LINFOMI CUTANEI (GILC)
Marco Lucioni1, Emilio Berti2, Aldo Maffi1, Luca Arcaini3,
Carlo Tomasini8, Pietro Quaglino8, Gaia Goteri4, Emanuela Boveri1,
Giorgio Croci1, Marta Nicola1, Antonio Ramponi9, Francesco Onida10,
Mauro Alaibac11, Stefano Ascani6, Roberta Riboni1, Sara Rattotti3,
Marcello Gambacorta7, Nicola Pimpinelli5, Marco Santucci5,
Marco Paulli1
remaining 20 (25%) had multiple lesions. PCFCL presented on the trunk (39 cases,
49%), the head and the neck (30 cases, 37%) and less frequently on the upper (7
cases, 9%) and the lower limbs (4 cases, 5%). Histologically, lymphomatous infiltrate consisted of follicular center cells, growing in a nodular (30/80), noduar and
diffuse (29/80) and purely diffuse (21/80) pattern. Immunophenotype was consistent with follicular origin in all the cases. FISH analysis documented the presence
of BCL2 translocation in 15/80 cases (18%), with a predominance of male patients
(14/15). All the cases carrying BCL2 translocation were also positive for bcl-2 by
immunohistochemistry.
Cutaneous relapse occurred in 23/80 patients (5 cases carrying BCL2 translocation). A single patient (BCL2 translocated) died of disease following systemic spread
53 months after diagnosis; Two patients died for other causes. At the last follow-up
62/77 (80%) patients were alive, disease-free; 15/77 (20%) patients were alive with
cutaneous disease.
Statystical analysis by Wilcoxon test, revealed that the presence of BCL2 translocation was associated with lower overall survival (p=0,020), lower event free survival
(p=0,004) and lower disease free survival (p<0,001).
Conclusions: At the best of our knowledge this is the widest series of PCFCL tested
for BCL2 translocation. Our data confirm that a fraction (18%) of PCFCL carry BCL2
translocation. Moreover the presence of this cytogenetic abnormality appears to
be associated with male sex, higher incidence of cutaneous relapses and worse
prognosis. These findings suggest that investigation of BCL2 translocation may be
relevant in the prognostic assessment of patients affected by PCFCL.
Keywords: Primary cutaneous follicular lymphoma; BCL2 translocation; FISH
analysis
[PP-LYMP-014]
EBV+ B CELL LYMPHOMAS (BCL) IN YOUNG PATIENTS
WITHOUT IMMUNODEFICIENCY
Alina Nicolae, Shahed Abdullah, Theresa Davies Hill,
Stefania Pittaluga, Elaine S. Jaffe
Hematopathology Section, National Cancer Institute, Bethesda, MD, USA
1
Anatomic Pathology Unit, Department of Molecular Medicine, University of Pavia, and Pathology
Unit, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy
Dermatology Section, Bicocca University Milan, Italy
3
Haemathology unit, University of Pavia/ IRCCS Policlinico San Matteo, Pavia
4
Pathology Department, Ospedali Riuniti, Ancona, Italy
5
Department of Surgery and Translational Medicine, Divisions of Dermatology and Anatomic
Pathology, University of Florence, Florence, Italy
6
Hematology Unit, IRCSS Santa Maria Nuova Hospital, Reggio Emilia, Italy
7
Department of Pathology, Niguarda Ca’ Granda Hospital, Milan, Italy
8
Dermatologic Clinic, Department of Medical Sciences, University of Turin; Anatomic Pathology Unit,
Hospital City of the Health and Science, Turin, Italy
9
Anatomic Pathology unit, Azienda Ospedaliera-Universitaria “Maggiore della carità”, Novara
10
Bone Marrow Transplant Center Hematology, Ospedale Maggiore IRCCS, Milan, Italy
11
Dermatologic Unit, University of Padova, Padova, Italy
2
Background: Primary Cutaneous Follicular Cell Lymphoma (PCFCL) is the most
common cutaneous B cell lymphoma and usually has an indolent clinical course (5
year overall survival 90%). PCFCL is a follicular centre cells neoplasm mainly consisting of centrocytic cells, characterized by expression of germinal center markers
(CD10 and Bcl6). In contrast to nodal follicular lymphoma, PCFCL usually lacks bcl-2
protein overexpression and the t(14;18)(q32;q21). However BCL2 gene translocation has been reported in a minority of cases (10-40%). Our aim was to investigate
the presence of t(14;18) by means of FISH in a large multicentric series of PCFCLs
and to assess possible differences in clinico-pathologic features and outcome between translocated and non translocated PCFCLs.
Material and methods: Eighty cases of PCFCL were collected from 9 different centres, members of Gruppo Italiano Linfomi Cutanei (GILC). Diagnostic biopsies of all
cases were reviewed by a collegial meeting of ten pathologists and dermatopathologists. Information about site, type and number of lesions was collected, as well as
follow-up data, including cutaneous relapse and extracutaneous spread.
Search for BCL2 gene translocation was performed by FISH analysis, using both
“dual color split signal” probes and “dual color dual fusion” probes, targeting
t(14;18)( q32;q21) translocation.
The presence of BCL2 translocation was correlated with Overall Survival (OS), Event
Free Survival (EFS) and Disease Free Survival (DFS) by statistical analysis.
Results: We collected 80 patients (M/F: 49/31) with a median age of 58 years (range
29-86). Sixty patients (75%) presented with single cutaneous lesions, whereas the
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The 2008 WHO classification included EBV+ diffuse large B-cell lymphoma (DLBCL)
of the elderly as a provisional entity. Rare EBV+ DLBCL cases in young pts have
been described, but it is uncertain if they resemble age related EBV+ DLBCL either
clinically or pathologically.
43 EBV+ BCL in young patients (age <=45) were identified in the authors’ files
from 2002 to date. Pts with immune deficiency were excluded. Other entities known
to be EBV+ were excluded: Burkitt, classical Hodgkin lymphoma, etc. Clinical features and outcome data were obtained. FFPE sections were stained for CD20, CD3,
CD15, CD30, CD79a, PAX5, Oct-2, Bob1, Bcl6, CD10, MUM1, EMA, IgD, CD21, LPM1,
EBNA2 and BZLF1, and EBER.
EBV+ BCL affected predominantly males (M:F=3.8:1), med age 23 (range 4-45).
93% presented with lymphadenopathy. 3 pts (7%) had only mediastinal disease.
Extranodal sites included lung (1), liver (3) and bone (3). 17/36 pts had B symptoms, 24/33 high LDH; BM+ in 9/32. Clinical stage in 32 cases was: I (3); II (12);
III (7) and IV (10). 53% had advanced stage. Available EBV serology indicated prior
infection (16/16) and/or virus reactivation (5 cases). 3 histological patterns were
identified: DLBCL-like (2); T/histiocyte-rich large B-cell lymphoma-like (THRBCL)
(35) and mediastinal gray zone lymphoma-like (GZL)(6). Except for DLBCL-like, all
others were polymorphic, rich in tumor cells mimicking HRS cells and/or LP cells
embedded in a prominent inflammatory background. None resembled polymorphic
posttransplant lymphoproliferative disorder (PTLD). 16/43 cases had prominent
sclerosis and 17/43 cases had focal or extensive areas of necrosis. By IHC, all but 1
expressed strong CD20. CD79a was + in 25/28, Oct-2 in 23/23 and Bob1 in 13/15
cases studied. PAX5 was strong in 16/31 and weak/variable in the remainder. CD30
was variably + in 36/43; only 3/43 were CD15 focally + (GZL-like cases). All cases
studied for MUM1 (39) were +, although 14/26 were Bcl6+. CD10 was neg. EBER
was present in >90% of tumor cells in all cases but 1. 40 cases studied for LMP1
were +, including the EBER neg one; 36/38 were EBNA2 neg (EBV latency type II);
2 cases were EBNA2+ (EBV latency type III). 3/39 had rare EBV+ tumor cells in
lytic phase, BZLF1+. Treatment and follow-up data were available for 33/43 pts.
All received chemotherapy, with R-CHOP the most common regimen (17/33), with
6 also receiving radiation. Two pts underwent autologous stem cell transplantation.
The median follow-up was 18 months (range 2-101 months). At the time of last
follow-up, 26/33 (78.8%) were in clinical remission with no evidence of disease, 4
pts were alive with disease (one recent case untreated) and only 3 pts (9%) died
EBV+ B cell lymphomas are not restricted to older pts, and occur in young pts without evident immunodeficiency. In contrast to the elderly they are more often nodal.
The most common histological pattern is THRBCL-like and rarely DLBCL-like. In
contrast to the elderly, PTLD-like lesions are not seen. Some resemble mediastinal
GZL, but are EBV+. Regardless of pathological features, they show an activated
B-cell immunophenotype, commonly express CD30 and have EBV latency type II.
Most of the pts respond to treatment and have a more favorable outcome than EBV+
DLBCL in the elderly.
INCOMPLETE CYTOKINESIS AND RE-FUSION OF SMALL
MONONUCLEATED HODGKIN CELLS LEADS TO GIANT
MULTINUCLEATED REED-STERNBERG CELLS
Keywords: EBV, diffuse large B-cell lymphoma, pediatric
Benjamin Rengstl1, Sebastian Newrzela1, Tim Heinrich1,
Christian Weiser1, Frederic B. Thalheimer2, Frederike Schmid1,
Kathrin Warner3, Sylvia Hartmann1, Timm Schroeder4, Ralf Küppers5,
Michael A. Rieger2, Martin Leo Hansmann1
1
Dr. Senckenberg Institute of Pathology, Medical School, Goethe-University of Frankfurt, 60590
Frankfurt am Main, Germany
2
LOEWE Center for Cell and Gene Therapy Frankfurt, Department of Hematology/Oncology, Medical
School, Goethe-University of Frankfurt, 60590 Frankfurt am Main, Germany
3
Department I of Internal Medicine, Medical School, University of Cologne, 50937 Cologne,
Germany
4
Stem Cell Dynamics Unit, Helmholtz Zentrum Munich, 85764 Neuherberg, Germany
5
Institute of Cell Biology (Cancer Research), Medical School, University of Duisburg-Essen, 45122
Essen, Germany
[PP-LYMP-015]
COPY NUMBER GAINS OF THE TNFRSF8 LOCUS IN
CD30-POSITIVE PERIPHERAL T-CELL LYMPHOMAS
Rebecca L. Boddicker, Andrew L. Feldman
Department of Laboratory Medicine, Mayo Clinic, Rochester, MN, USA
Background: CD30 is a transmembrane tumor necrosis factor receptor superfamily member encoded by the TNFRSF8 gene on 1p36.22. CD30 is constitutively expressed in a subset of peripheral T-cell lymphomas (PTCLs), particularly anaplastic
large T-cell lymphomas (ALCLs). While CD30 expression is induced physiologically
upon activation of normal T-cells, the mechanisms for its constitutive expression
in PTCLs remain incompletely understood. Recently, we identified a novel positive
feedback loop in PTCL cells, whereby the transcription factor interferon regulatory
factor-4 (IRF4/MUM1) induced CD30 expression, which then led to increased IRF4
expression via NF-B. Interestingly, we found that a subset of PTCLs co-expressing
IRF4 and CD30 had copy number gains of the IRF4 locus. In this study, we examined
the frequency and distribution of gains of TNFRSF8 in PTCLs.
Methods: Tissue microarrays containing paraffin-embedded PTCLs from 126 patients diagnosed by WHO criteria were analyzed by fluorescence in situ hybridization
(FISH) using probes that hybridized to TNFRSF8 (red) and the PBX1 locus on 1q23.3
(green). Cases included 43 ALCLs (24 ALK-negative, 13 ALK-positive, and 6 primary
cutaneous); 41 PTCLs, not otherwise specified (NOS); 20 angioimmunoblastic T-cell
lymphomas (AITLs), and 22 other PTCLs. Most cases had been previously analyzed
by FISH for IRF4 and by immunohistochemistry for CD30, IRF4, and Ki67.
Results: Of 126 cases analyzed, 18 (14%) had gains of TNFRSF8 (copy number
range, 3-15; mean, 5.8; median, 4.5). Copies of TNFRSF8 outnumbered copies of
PBX1 in 11/18 cases, and were associated with corresponding gains of PBX1 in
7. Gains of TNFRSF8 were seen in 29% of CD30-positive PTCLs (>=30% tumor
cells positive) versus 8% of CD30-negative PTCLs (p=0.0066, Fisher’s exact test).
PTCLs with gains of TNFRSF8 also had more CD30-positive cells (mean ± S.D., 62
± 46% versus 30 ± 44%; p=0.0087, Wilcoxon test). Gains of TNFRSF8 were most
frequent in ALK-negative and primary cutaneous ALCLs (33% each, versus 8% in
ALK-positive ALCLs; p=0.12). They were seen in 12% of PTCLs, NOS, and were
absent in all AITLs and most other PTCLs tested. PTCLs with gains of TNFRSF8 also
had more tumor cells positive for IRF4 (58 ± 44% versus 23 ± 35%; p=0.0029)
and for Ki67 (55 ± 29% versus 37 ± 30%; p=0.024). Among 23 cases with gains of
either TNFRSF8 or IRF4, only one case had gains of both loci.
Conclusions: Copy number gains of the TNFRSF8 locus are associated with CD30
expression in PTCLs. Cases with gains of TNFRSF8 also showed increased expression of both IRF4 and the proliferation marker, Ki67. Gains of TNFRSF8 showed
minimal overlap with gains of IRF4. Presence of these copy number abnormalities
may contribute to constitutive co-expression of CD30 and IRF4 and, based on data
by us and others, to proliferation in some PTCLs. Further inquiry into mechanisms
regulating CD30 expression in PTCLs lacking these abnormalities is merited.
Multinucleated giant tumor cells can be frequently observed in tissue sections of
lymphoma patients. In classical Hodgkin lymphoma (HL) as well as HL of T-cell
origin, these so-called Reed-Sternberg (RS) cells are pathognomonic for the
disease. Despite the well-described disease-promoting functions of multinucleated RS cells, their development still remains obscure. It was postulated that RS
cells arise from mononucleated Hodgkin cells via endomitosis. Conversely, continuous single cell tracking of HL cell lines by long-term time-lapse microscopy
showed that cell fusion is the main route of RS cell formation (Fig. 1). In contrast
to growth-induced formation of giant Hodgkin cells, fusion of small mononuclear
cells followed by size increase gives rise to giant RS cells. Importantly, we nearly
exclusively observed fusion of cells originating from the same ancestor, termed
re-fusion. In the majority of cases, re-fusion of daughter cells was preceded by
an incomplete cytokinesis, visualized by a microtubule bond between the cells
(Fig. 2). We confirm at the level of individual tracked cells that giant Hodgkin and
RS cells have little proliferative capacity, further specifying small mononuclear
Hodgkin cells as the proliferative compartment of the HL tumor clone. In addition,
sister cells showed a shared propensity for re-fusion, which provides evidence
of early RS cell fate commitment. Thus, RS cell generation is neither due to cell
fusion of unrelated Hodgkin cells nor to endomitosis, but is mediated by re-fusion
of daughter cells that underwent mitosis. This surprising finding indicates the
existence of a novel mechanism for the generation of multinuclear RS cells, which
might have implications beyond HL, as RS-like cells are abundant in several other
lymphoproliferative diseases.
[PP-LYMP-016]
of disease. The pts with refractory disease showed either systemic disease and/or
mediastinal involvement.
Keywords: Hodgkin lymphoma, incomplete cytokinesis, cell fusion
Figure 1. RS cells primarily arise from re-fusion of daughter cells.
Keywords: CD30, copy number abnormalities, T-cell lymphoma
Figure 2. Incomplete cytokinesis precedes re-fusion of HRS daughter cells.
| 17-22 October 2014
| EAHP - 2014
|
85
th
[PP-LYMP-017]
THE ROLE OF ICOS IN THE PATHOGENESIS OF AITL
Frederike Schmid1, Benjamin Rengstl1, Anna Janton1,
Christian Weiser1, Sylvia Hartmann1, Kathrin Warner2,
Martin Leo Hansmann1, Sebastian Newrzela1
1
Dr. Senckenberg Institute of Pathology, University Medical Center Frankfurt, Frankfurt am Main,
Germany
2
Department of Medicine I, Cologne University, Cologne, Germany
Angioimmunoblastic T-cell lymphoma (AITL) is one of the most frequent mature
T-cell lymphomas. In the recent years much effort has been made to elucidate the
molecular mechanisms that drive the pathogenesis of AITL. One major step was the
discovery that the neoplastic cells are most likely derived from follicular T helper
cells (TFH) (de Leval L, et al. Blood 2007; Piccaluga PP, et al. Cancer Research
2007). These cells are characterized by the expression of CD4, ICOS, CXCR5 and
Bcl6. Expression of ICOS is pivotal for the formation of TFH (Akiba H, et al. JI 2005).
Furthermore, ICOS can protect the cells against apoptosis (Hutloff A, et al. Nature
1999), which might be a pathway promoting transformation. Currently, there is no
ICOS overexpressing mouse model available, but overexpression of ICOS induces an
AITL-like phenotype in the Roquinsan mouse model (Ellyard JI, et al. Blood 2012).
This suggests a tumor-promoting role of ICOS. The aim of the presented study was
to assess a potential oncogenic role of ICOS in the generation of T-cell lymphoma.
To address the role of ICOS in promoting lymphoma formation ICOS and GFP control
gene expressing murine hematopoietic stem cells were transplanted into immunedeficient recipients. However, no malignancy development was observed in recipient animals. Analysis of sacrificed animals revealed that ICOS overexpression solely
resulted in a weak increase in TFH cellularity. On the contrary, the population of
germinal center B cells (GCBC) was strongly enlarged. This leads to the interpretation that even a slight increase in ICOS expression can strongly promote proliferation
of GCBC and is inline with the fact that AILT is often accompanied by strong BC
proliferation in the lymph node.
Under the experimental conditions in our mouse model ICOS did not show oncogenic potential. However, ICOS could be a factor contributing to lymphoma formation by acting as a co-oncogene. In a new setting CD4+ T cells were enriched or
depleted of the ICOS+ fraction and then transduced with the oncogene NPM-ALK.
The latency of tumor onset was strongly reduced in animals transplanted with the
ICOS+ T-cell fraction, thus ICOS+ T cells seem to be more susceptible to transformation than ICOS- T cells.
Our study suggests that overexpression of ICOS cannot induce lymphoma in our
mouse models. However, ICOS+ T cells seem to be transformed more easily and can
accelerate tumor formation. Understanding the co-oncogenic role of ICOS might improve our understanding of the molecular pathways involved in the generation of AITL.
Keywords: Angioimmunoblastic T cell lymphoma, ICOS, NPM-ALK
Besides negativity for CD10/CD23/bcl6/cyclinD1, an extended panel of MNDA1/
MUM1/IGD/ HCAM was used for diagnosis and relevant clinical details and follow
up was obtained.
The median age group of the patients included in this study was 56 years with age
ranging from 35-76 years. Out of the 67 cases of NMZL, on review, only 20 could
be reclassified as NMZL which included a case of pediatric NMZL. The other cases
were reclassified as SLL/CLL (small lymphocytic lymphoma) with aberrant immunophenotype, follicular lymphoma with marginal zone differentiation (FLMZD), splenic
marginal zone lymphoma(SMZL) and reactive pathologies(Castleman’s disease, and
IgG4 related disease). None of the other low grade lymphoma differentials other
than NMZL presented with stage I disease. The 20 cases of NMZL showed varied
morphologic and cytological features. Plasma cells were consistently identified in
all cases of NMZL giving a clue to diagnosis. The major reason of misdiagnosis of
SLL/CLL was the atypical morphology and immunohistochemistry. We observed that
flow cytometry helped in picking up these misdiagnoses cases. Failure to include
bcl6 in immunopanel and lack of follicular growth lead to misdiagnosis of FLMZD.
The immunopanel of MNDA, MUM-1, CD44, IgD was not helpful to delineate one
disease from the other. A survival analysis revealed that compared with the misdiagnosed entities NMZL had superior prognosis.
Conclusion: NMZL is indeed a wastebasket diagnosis and hence diagnosis should
not be made without a flow cytometry work up in cases with marrow involvement
and full clinical picture. Accurately defined NMZL indeed had excellent prognosis
contrary to other differential diagnosis.
Keywords: Nodal marginal lymphoma
[PP-LYMP-019]
THE PROGNOSTIC VALUE OF MYC, BCL2 AND BCL6
TRANSLOCATIONS AND PROTEIN EXPRESSIONS IN YOUNG
PATIENTS WITH HIGH-RISK DIFFUSE LARGE B-CELL
LYMPHOMA TREATED WITH R-CHOEP
Mette Ø. Pedersen1, Anne O. Gang2, Estrid Høgdall1, Helle Knudsen1,
Anne F. Lautitzen1, Signe L. Nielsen1, Tobias W. Klausen2,
Michael Pedersen3, Peter Brown3, Peter Nørgaard1
1
Department of Pathology, Herlev Hospital, Denmark
Department of Hematology, Herlev Hospital, Denmark
3
Department of Hematology, Rigshospitalet, Denmark
2
Background: In young patients with high-risk diffuse large B-cell lymphoma
(DLBCL) treatment with R-CHOEP (R-CHOP + etoposide) has been associated with
improved outcome. Whether established prognostic markers in R-CHOP treated patients are prognostic in R-CHOEP treated patients remain to be investigated. In addition predictive markers for response to R-CHOEP need to be investigated.
Methods: A Danish population based cohort of 140 young (age 18-60) patients with
high-risk (2 >= additional risk factors including advanced stage, elevated s-LDH,
and performance status >1) primary DLBCL diagnosed between 2004 and 2008
was investigated. Patients were treated with R-CHOP (n=84) or R-CHOEP (n=56).
Formalin fixed paraffin embedded tumor tissue specimens were analysed for MYC, BCL2- and BCL6- protein expression by semiquantitative immunohistochemistry
(IHC) and genetic translocations by FISH.
Figure 1. Histological section. Representative histological sections of animals
transplanted with EGFP and NPM-ALK transduced T cells.
[PP-LYMP-018]
NODAL MARGINAL ZONE LYMPHOMA-UPFRONT DIAGNOSIS
WITHOUT WORK UP IS OFTEN A WASTEBASKET DIAGNOSIS
Katha Kanthe, Tanuja Shet, Sridhar Epari, Manju Sengar, Hari Menon
Tata Memorial Hospital, Parel, Mumbai
Nodal marginal zone lymphoma is often a diagnosis of exclusion when other low
grade lymphomas are excluded. This was a retrospective analysis of 67 cases of
nodal marginal zone lymphoma (NMZL) with a view to asses accuracy of diagnosis
and define immunophenotypic features of NMZL.
86
| EAHP - 2014
| 17-22 October 2014
Results: MYC protein expression >= 40% was seen in 67/106 patients (71%), BCL2
protein expression > 0 and BCL2 protein expression >= 70% was seen in 106/138
(77%) and 81/117 patients (69%) respectively. BCL6 expression >= 30% was seen
in 96/114 patients (84%). Concurrent expression of MYC>=40% and BCL2>=70%
(IHC DH) was seen in 50/106 patients (47%). Concurrent MYC>=40%, BCL2 >0 or
BCL6<30% (TH score 2-3) was seen in 61/103 patients (59%)
MYC, BCL2 and BCL6 translocation was seen in 14/104 (13%), 31/104 (30%) and
27/104 (26%) patients respectively. Concurrent MYC BCL2/BCL6 translocation (DH)
was seen in 8/102 (8%) patients.
MYC over-expression was not associated with reduced progression free survival
(PFS) in either R-CHOP or R-CHOEP treated patients. BCL2 expression (>0%) and
BCL2 overexpression (>=70%) was associated with reduced PFS in R-CHOP (HR:
0.3; 95%CI:0.1-0.9; p=0.03 and HR: 0.4; 95%CI: 0.2-0.9; p=0.02) - but not in
R-CHOEP treated patients (HR: 0.9; 95%CI:0.3-3.2; p=0.9 and HR: 0.5; 95%CI: 0.11.9; p=0.3). IHC DH was associated with a trend towards reduced PFS in R-CHOP
- but not in R-CHOEP treated patients (HR: 0.6; 95%CI:0.3-1.1; p=0.08 and HR: 1.2;
95%CI: 0.4-4.0; p=0.7 respectively). TH score 2-3 was associated with reduced PFS
in R-CHOP - but not in R-CHOEP treated patients (HR: 0.4; 95%CI:0.2-0.9; p=0.015
MYC, BCL2 and BCL6 translocations were not prognostic markers with respect to
PFS in either R-CHOP or R-CHOP treated patients. DH translocations were too few to
perform meaningful statistical analyses.
THE SURVIVAL OF PATIENTS WITH T(14;18)-NEGATIVE
FOLLICULAR LYMPHOMAS IS NOT DIFFERENT AS COMPARED
TO PATIENTS WITH T(14;18)-POSITIVE FOLLICULAR
LYMPHOMAS
Conclusions: BCL2 expression, IHC DH and TH score 2-3 had prognostic value in
R-CHOP treated patients but this was not seen in R-CHOEP treated patients in this
study suggesting the need for novel prognostic markers in this group of patients.
Neither BCL2 expression, IHC DH nor TH score 2-3 were however predictive markers
for response to treatment with R-CHOEP in this cohort of patients. This could possibly be due to the quite small patient-cohort investigated in this study and examination of larger patient cohorts could be advantageous in future studies.
MYC BCL2/BCL6 DH translocations were too few to perform meaningful statistical
analyses and whether DH translocation will retain a prognostic value in R-CHOEP
treated patients also remain to be investigated in larger patient cohorts. A possible
predictive value of DH translocation also needs further investigation.
Keywords: diffuse large B-cell lymphoma, MYC, BCL2
[PP-LYMP-020]
DEFINING HODGKIN LYMPHOMA ON A MOLECULAR BASIS
Sylvia Hartmann1, Claudia Döring1, Christina Jakobus1,
Benjamin Rengstl1, Sebastian Newrzela1, Thomas Tousseyn2,
Xavier Sagaert2, Maurilio Ponzoni3, Fabio Facchetti4,
Christiane De Wolf Peeters2, Christian Steidl5, Randy Gascoyne5,
Ralf Küppers6, Claudio Agostinelli7, Pier Paolo Piccaluga7,
Stefano Pileri7, Martin Leo Hansmann1
1
Dr. Senckenberg Institute of Pathology, Goethe University, Frankfurt am Main, Germany
Department of Pathology, University Hospitals K.U.Leuven, Leuven, Belgium
3
Unit of lymphoid malignancies, Department of Pathology, Scientific Institute San Raffaele, Milan,
Italy
4
Department of Pathology, University of Brescia, Brescia, Italy
5
Department of Pathology and Laboratory Medicine and the Centre for Lymphoid Cancer, British
Columbia Cancer Agency, University of British Columbia, Vancouver, Canada
6
Institute of Cell Biology (Cancer Research), Faculty of Medicine, University of Duisburg-Essen,
Essen, Germany
7
Department of Experimental, Diagnostic and Specialty Medicine, Haematopathology Section,
S Orsola-Malpighi Hospital, University of Bologna, Bologna, Italy
2
Hodgkin lymphoma is characterized by a low tumor cell content in the infiltrate.
Differential diagnoses include ALK- anaplastic large cell lymphoma (ALCL) for classical Hodgkin Lymphoma (cHL) and T cell/histiocyte rich large B cell lymphoma
(THRLBCL) for nodular lymphocyte predominant Hodgkin Lymphoma (NLPHL). To
be able to differentiate these cases we reevaluated existing and generated new
gene expression data of microdissected tumor cells of the respective lymphoma
types. ALK- ALCL and cHL could be well differentiated applying immunohistochemical stainings for four differentially expressed genes (CD83, MDC/CCL22, STAT3
and TUBB2B) overexpressed in cHL. These genes have also been found to be expressed in antigen-presenting cells, indicating that Hodgkin-Reed-Sternberg (HRS)
cells maintain functions usually observed in reactive B cells. In contrast, tumor
cells of NLPHL and THRLBCL showed no consistent differences in gene expression. Overexpressed genes, which were validated on protein level, were detected
in a subset of tumor cells of typical and variant NLPHL as well as in THRLBCL. We
therefore conclude, that HRS cells in cHL show fundamental functional differences
compared to tumor cells of ALK- ALCL, whereas NLPHL and THRLBCL may represent
a true continuum and endpoints of a spectrum of the same disease.
Keywords: Hodgkin lymphoma, anaplastic large cell lymphoma, T cell/histiocyte rich
large B cell lymphoma
Ellen Leich1, Martin Wartenberg1, Michael Unterhalt2,
Reiner Siebert3, Heike Horn4, Wolfram Klapper5,
Heinz Wolfram Bernd6, Michael Hummel7, Harald Stein8,
Sylvia Hartmann9, Peter Möller10, Wolfgang Hiddemann2,
German Ott4, Andreas Rosenwald1
1
Institute of Pathology, University of Würzburg, Germany
Department of Internal Medicine III, University of Munich, Germany
Department of Human Genetics, University of Kiel, Germany
4
5
Institute of Pathology, Hematopathology Section and Lymph Node Registry, University Hospital
Schleswig-Holstein, Campus Kiel, Germany
6
Institute of Pathology, University Hospital Schleswig-Holstein, Campus Lübeck, Germany
7
Institute of Pathology, Campus Benjamin Franklin, Charité Universitätsmedizin, Berlin, Germany
8
Pathodiagnostik Berlin, Germany
9
Institute of Pathology, University Hospital Frankfurt, Germany
10
Institute of Pathology, University Hospital Ulm, Germany
2
3
Approximately 85% of follicular lymphomas (FL) carry the translocation t(14;18)
that contributes to FL pathogenesis. In contrast, pathogenetic features and relevant
clinical parameters of ~15% of FL that lack the t(14;18) are largely unknown. The
aim of this study was therefore to better define molecular and clinical features of
t(14;18)-negative FL.
In a tissue microarray (TMA) format, we studied the presence or absence of the
t(14;18) as well as breaks in the BCL6 and MYC loci by fluorescence in situ hybridization (FISH) in a large cohort of FL stages III/IV patients treated within the German
Low Grade Lymphoma Study Group (GLSG). The protein expression status of BCL2,
BCL6, MUM1, CD10, p53 and Ki-67 was determined by immunohistochemistry as
well. Altogether, 539 FL patients (mostly grade 1/2 tumors) treated with either MCP,
CHOP or R-CHOP were investigated. The analysis focused on the comparison of
biological and clinical features between t(14;18)-positive and t(14;18)-negative FL.
[PP-LYMP-021]
and HR: 0.8; 95%CI: 0.2-2.8; p=0.74). There was no statistical significant interaction between treatment modality and BCL2 expression, IHC DH or TH score 2-3.
Breaks affecting the BCL2 locus were detected in 363 of 422 evaluable FL specimens (86%), which is in line with previous findings. BCL6 breaks were observed
in 45/443 FL (10%) and MYC breaks in 10/424 FL (2%). There was no enrichment
of BCL6 or MYC breaks in the t(14;18)-positive or t(14;18)-negative FL subgroup.
Comparing immunohistochemical features between the two subgroups, BCL2
(measured with three different BCL2 antibodies) was expressed in 99% of t(14;18)positive FL, but also in 77% of t(14;18)-negative FL, a number which is significantly
higher than that reported in previous studies. This relatively high number of BCL2
expression in the group of t(14;18)-negative FL was observed irrespective of the
BCL2 antibody used. Of note, a genomic gain of the BCL2 locus that could account
for this finding was detected in only a small subset of analyzed cases and was
not restricted to FL without t(14;18). 17 of 51 tumors (33%) among the t(14;18)negative FL were CD10-negative, while only 27 out of 352 FL (8%) showed this
feature within the t(14;18)-positive group (p<0.05) demonstrating a clear enrichment of CD10-negative cases within t(14;18)-negative FL. No significant differences
in the expression of other immunohistochemical markers employed were detected
between the subgroups.
Since different treatment arms and FLIPI scores were evenly distributed between
t(14;18)-positive and t(14;18)-negative FL, we included all patients for the analysis
of clinical parameters. Most importantly, the median overall survival after diagnosis
did not differ between the group of t(14;18)-negative and t(14;18)-positive FL patients (~9.4 vs. ~8.6 years). Likewise, no significant difference for time to treatment
failure, FLIPI score and ECOG status was observed. There was only a slight tendency
towards an inferior initial response to therapy in the group of t(14;18)-negative FL.
In conclusion, the results of the current study in a large cohort of FL patients treated
within prospective clinical trials of the GLSG suggest a high degree of similarity
between t(14;18)-positive and t(14;18)-negative FL with regard to chromosomal
breaks in the BCL6 and MYC loci as well as to the immunohistochemical marker
profile with the exception of an enrichment of CD10-negative FL among t(14;18)negative FL. Moreover, FL with and without t(14;18) do not appear to show differences in their clinical behaviour supporting the notion that they represent two
subgroups within the same entity.
Keywords: follicular lymphoma, t(14;18), CD10
| 17-22 October 2014
| EAHP - 2014
|
87
th
[PP-LYMP-022]
[PP-LYMP-023]
THE RELATIONSHIP BETWEEN OVERT AND IN SITU LYMPHOMA:
A RETROSPECTIVE STUDY OF CASES OF FOLLICULAR AND
MANTLE CELL LYMPHOMA
BCL-2 NEGATIVE FOLLICULAR LYMPHOMA: AN EFFORT TO
IMPROVE OUR UNDERSTANDING OF THIS SPECIAL SUBTYPE OF
FOLLICULAR LYMPHOMAS
Larissa Sena Teixeira Mendes, Ayoma Attygalle, Andrew Wotherspoon
Georgia Levidou1, Claudio Agostinelli2, Elena Sabattini2,
Simona Righi2, Simone Sabbioni2, Riccardo Panzacchi2,
Ayse U. Akarca3, Efstratios Patsouris1, Teresa Marafioti3,
Stefano A. Pileri2
Royal Marsden Hospital - Histopathology Department. London, UK
Follicular and mantle cell lymphoma in situ are characterised by the presence of
cells with the characteristic immunophenotype and genotype of the overt lymphoma in the appropriate architectural niche in asymptomatic patients. Follicular
lymphoma in situ (FLIS) is diagnosed when germinal centre cells are present with
strong expression of bcl-2 protein within a proportion of follicle centres in an otherwise unremarkable reactive lymph node. Mantle cell lymphoma in situ (MCLIS)
is characterised by cyclinD1 positive cells within the inner part of the mantle in
similar unremarkable reactive nodes. In general neither can be diagnosed without
immunohistochemical studies as the morphological changes are extremely subtle.
While it is known that only a small proportion of patients with FLIS will progress to
overt follicular lymphoma, it is unknown whether all cases of follicular lymphoma
are preceded by in situ disease. The situation with MCLIS is even less certain as
this is a very rare finding. In order to study this in more detail we have searched the
diagnostic histopathology archive at the Royal Marsden Hospital for patients with
follicular lymphoma diagnosed from 1996 to 2013 that had previously undergone
resections including lymph nodes, tonsil or appendix.
Twelve cases of follicular lymphoma were available for review. For the MCL cases, only
one of the resections included lymph nodes. These resections were retrieved from the
surgical pathological archives and stained with bcl-2 protein for patients with follicular
lymphoma and cyclinD1 in patients with mantle cell lymphoma. If initial staining for
bcl-2 was negative a second stain was performed with the bcl-2 E17 clone.
Of the 12 cases of follicular lymphoma 10 cases where shown to have pre-existing
FLIS. The single case of MCL showed in situ lymphoma in the original lymph node
resected. The average interval between original resection with FLIS and the detection of overt FL was 97.5 months (range 06-264 months). The interval between first
resection with MCLIS and overt MCL was 240 months. (Table 1)
While the numbers in this study are small our findings suggest that the vast majority of patients who develop follicular lymphoma will have had pre-existing FLIS
although some cases may develop de novo. With mantle cell lymphoma the situation
may be the same, but no definite conclusion can be inferred from our single case.
Keywords: In situ lymphoma, follicular lymphoma in situ, mantle cell lymphoma in situ
Table 1. Overt and in situ lymphoma relationship: cases overview
Follicular Lymphoma (FL)
Original Bps
Overt FL
Patient
Gender
Dx - Sample
Year
FLIS
(bcl-2)
Grade
Year
Interval
(months)
01
Female
IDC – LN
1996
+
1-2
2002
72mo
02
Female
IDC – LN
2007
+
1-2
2013
192mo
03
Female
MM – LN
2005
+
1
2011
72mo
04
Female
IDC – LN
1997
-
3B*
2001
48mo
05
Female
IDC – LN
1985
+
3B*
1997
144mo
06
Female
IDC – LN
1999
+
1
2007
96mo
07
Male
Stomach CA – LN
2005
+
1
2012
84mo
08
Male
MM – LN
1989
+
1
2011
264mo
12mo
09
Male
SCC Met – LN
2001
-
1-2
2002
10
Female
Colon CA – Apd
1995
+
1
1996
6mo
11
Male
PA – SG
2006
+
3A
2013
84mo
12
Female
Warthin Tu – SG
2005
+
1-2
2013
96mo
1
Section of Haematopathology, Department of Haematology and Oncological Sciences Seràgnoli,
S Orsola-Malpighi Hospital, University of Bologna, Bologna, Italy
3
Department of Histopathology, University College Hospital, London, UK
2
Background: The hallmark of follicular lymphoma (FL) is t(14;18)(q32;q21) chromosomal translocation, being identified in 65-85% of cases. This rearrangement leads
to aberrant expression of BCL-2 protein in the majority of cases. It is well recognised
that a minority of FL cases lack t(14;18) and are negative for BCL-2 expression.
Stathmin expression has been previously identified as a potential diagnostic marker
for FL. The paucity of information regarding BCL-2 negative (in immunohistochemical and molecular terms) FL cases prompted us to undertake this investigation in
an effort to elucidate the clinical, pathological and molecular features as well as
Stathmin expression in this subgroup of cases.
Materials-Methods: Fifty one cases with BCL-2 (-) lymph node FLs for which paraffin embedded tissue was available were included in the present investigation.
BCL2 staining (clone 124, E17 and SP66) as well as BCL-2 FISH analysis were performed in all cases before enrolment in the study. Stathmin expression was evaluated immunohistochemically in 36 cases. All cases were also investigated with a
panel of immunohistochemical markers (BCL-6, CD10, IRF-4/MUM-1, CD30, CD21,
CD23, IgM, IgG, IgD, IgA, CD68, PD-1, FOXP-3, p53, MYC and IRTA-1) and with FISH
analysis for the presence of BCL-6 and MYC rearrangement (8q24). The pattern of
Stathmin expression was compared with that observed in 19 reactive lymph nodes.
GEP analysis in silico was also performed in order to evaluate the immunohistochemical results.
Results: The patients in our cohort tended to be <60 years old (64%) whereas
there was a female predominance, which seems to be rather more prominent compared to what is reportedly observed in classical BCL-2 (+) FL. The majority of the
cases involved inguinal lymph nodes (60%) and had a higher histological grade
(59% grade IIIA, 22% grade IIIB). Five cases were CD10 negative, whereas 6 were
IRF-4/MUM-1 positive. MYC or p53 expression was not observed. 15% of the cases
displayed an alternative molecular change, namely BCL-6 translocation. Stathmin
expression was observed in all cases. In 32 cases the range of positivity was between 10 and 75%, while in reactive lymph nodes >75% of germinal centre cells
were Stathmin positive (p<0.0001). Only in 4 cases (11%) Stathmin expression was
comparable to that observed in normal lymph nodes. In those 4 cases positivity
for MUM-1/IRF-4 was also observed, a finding that could possibly indicate B cell
activation, although CD30 expression was not observed. Moreover, the remaining
immunohistochemical analysis (FOXP3, PD1, CD68) did not reveal any difference in
the microenvironment between the 4 cases displaying increased (>75%) expression
of Stathmin and the rest 32 cases. Interestingly, GEP analysis in silico showed that
Stathmin is mainly expressed in centroblasts (CB) and centrocytes (CC) in contrast
to memory or naïve B cells whereas it seems to be downregulated in follicular lymphomas when compared to normal CC and CB.
Conclusion: BCL-2 (-) FL seems to be an entity with distinctive features, i.e. younger
female patients with inguinal site prevalence and alternative molecular aberrations.
Stathmin expression is invariably observed in BCL-2 (-) FLs, though in lower levels
when compared to normal lymph nodes and could therefore serve as an additional
marker for its differential diagnosis from follicular hyperplasia. Studies on new germinal centre markers are ongoing and will be presented in the meeting.
Keywords: BCL-2 negative FL, Stathmin, differential diagnosis from follicular
hyperplasia
Mantle Cell Lymphoma (MCL)
Original Bps
Patient
Gender
Dx - Sample
Year
MCLIS
(cyc-D1)
Overt MCL
Year
Interval
(months)
01
Male
MM - LN
1988
+
2008
240mo
Bps: biopsy; FLIS: follicular lymphoma in situ, Dx: diagnosis; IDC: invasive ductal carcinoma/ Breast;
LN: lymph node; CA: Carcinoma; MM: melanoma; Apd: appendix; SCC met: metastatic squamous cell
carcinoma; PA.: pleomorphic adenoma; SG: salivary gland; Tu: tumour; MCLIS: mantle cell lymphoma
in situ.
88
| EAHP - 2014
| 17-22 October 2014
VALIDATION OF MULTIPLE IMMUNOHISTOCHEMICAL
ALGORITHMS FOR ASSIGNING DIFFUSE LARGE B-CELL
LYMPHOMA SUBTYPES USING A CLINICAL MOLECULAR DLBCL
SUBTYPING ASSAY
BCL6 REGULATION BY MICRORNA CONTROL TFH PHENOTYPE
IN PTCL
Angela M. B. Collie1, Jork Nolling2, Kiran M. Divakar2, Jeffrey J. Lin1,
Brian T. Hill3, Mitchell R. Smith3, Lilly I. Kong2, Elena A. Manilich4,
Eric D. Hsi1
1
Department of Laboratory Medicine, Cleveland Clinic, Cleveland, Ohio, USA
PrimeraDx, Mansfield, Massachusetts, USA
3
Department of Hematologic Oncology and Blood Disorders, Cleveland Clinic, Cleveland, Ohio, USA
4
Department of Colorectal Surgery, Cleveland Clinic, Cleveland, Ohio, USA
Rebeca Manso1, Nerea Martínez Magunacelaya2, Cristina Chamizo1,
Federico Rojo1, Jesús García Foncillas3, Miguel Ángel Piris2,
Socorro María Rodríguez Pinilla1
1
Pathology Department, IIS-Fundación Jiménez Díaz, UAM, Madrid, Spain
Laboratorio de Genómica del Cáncer, IDIVAL, Santander, Spain
3
Translational Oncology Division, Oncohealth Institute, IIS-Fundación Jiménez Díaz, UAM, University
Hospital “Fundación Jiménez Díaz”, Madrid, Spain
2
2
Diffuse large B-cell lymphoma (DLBCL) can be classified into two molecular subtypes based on normal developmental B-cells, the putative cell of origin, using
microarray gene expression profiling (GEP). These two molecular subtypes are
germinal center B-cell-like (GCB) and activated B-cell-like (ABC) DLBCL. Accurate
molecular subtype classification is crucial for prognostic, and potentially therapeutic, reasons. Multiple immunohistochemical (IHC) algorithms have been developed as surrogate methods for determining molecular subtype. Ideally, these
methods should be validated in clinical laboratories using a molecular test before
implementation. Therefore, we validated four IHC algorithms in DLBCL cases from
our institution against our clinical DLBCL molecular subtyping assay, which uses
formalin-fixed paraffin-embedded tissue (FFPE)(Collie AMB et al. Br J Haematol.
In press, 2014).
95 de novo DLBCL samples from patients who were subsequently treated with
R-CHOP were identified. Overall survival and International Prognostic Index (IPI)
score were collected. Immunohistochemical stains using antibodies for BCL6,
CD10, FOXP1, GCET1, LMO2, and MUM1 were performed on a tissue microarray. The cases were classified according to several published IHC algorithms into
cell of origin subtype: germinal center-B-cell-like (GCB) or non-GCB (NGC) DLBCL
(Hans CP et al. Blood. 2004; 103: 275-282. Choi WW et al. Clin Cancer Res. 2009;
15: 5494-5502. Meyer PN et al. J Clin Oncol. 2011; 29: 200-207. Visco C et al.
Leukemia. 2012; 26: 2103-13.). The multiplex PCR-based ICEPlex assay, utilizing a 14-gene panel, was performed on a single 10-μm slice of FFPE from 63 of
the DLBCL samples, and molecular subtype was assigned based on the Wright
classification.
The median age of the DLBCL cohort was 63 years, and 57% of patients had stage
III or IV disease. Median follow-up was 56 months. For all four IHC algorithms, overall
survival was significantly shorter for patients with the NGC subtype compared to
the GCB subtype (p<0.05). Compared to the DLBCL molecular subtyping assay, the
Hans algorithm correctly assigned subtype in 88.5% of GCB cases and 94.6% of
ABC cases, the Choi algorithm correctly assigned subtype in 96.2% of GCB cases
and 97.3% of ABC cases, the Tally algorithm correctly assigned subtype in 92.3%
of GCB cases and 97.3% of ABC cases, and the Visco-Young algorithm correctly assigned subtype in 84.6% of GCB cases and 100% of ABC cases.
We have successfully validated four published IHC algorithms for DLBCL subtyping
in an independent cohort from our institution. All four IHC algorithms showed correlation with overall survival and high concordance with our clinical DLBCL molecular subtyping assay using FFPE specimens. Validation of IHC algorithms for DLBCL
subtyping at any clinical institution is critical and can be effectively accomplished
using molecular-based assays.
INTROCUCTION: Peripheral T-cell lymphomas (PTCLs) are neoplasms derived from
mature post-thymic T-cell lymphocytes that account for approximately 12% of all
lymphoid neoplasms. They are subclassified as either predominantly nodal or extranodal based on their main site of involvement. Among nodal PTCLs, angioimmunoblastic T-cell lymphoma (AITL) and PTCL-NOS are the most frequent tumor
types. Gene expression profiling studies have revealed that PTCL-NOS are derived
from activated T-cells, with AITL harboring a gene-expression signature suggesting
derivation from follicular helper T-cells. Follicular Th cells (TFH) are a new characterized CD4 lineage whose major function is to help B cells from germinal centres
(GCs). TFH cells have an activated effector T cell phenotype and express elevated
levels of ICOS, PD1, CXCL13, CXCR5 and CXCR4. The BCL6 transcriptional repressor
protein is upregulated in Tfh cells and is considered a master regulator for the TFH
cell lineage.
Aim: Probe whether microRNAs could regulate the TFH phenotype among PTCLs by
regulating BCL6 expression. Both previously reported target mirs of the BCL6 gene
as well as predicted target of the gene found in several databases (pictar, Miranda,
mirbase) were tested.
[PP-LYMP-025]
[PP-LYMP-024]
Material-Methods: Twenty-seven PTCL frozen samples were hybridized to obtain
both gene and microRNA expression data using arrays. Associations between genes
and microRNAs were carried out with SPSS 12.0 (SPSS Inc., Chicago, IL) using the
Pearson correlation exact test. Significant genes and microRNAs were further validated using RT-PCR in a series of 100 paraffin-embedded PTCL-NOS.
Results: A direct statistical relationship between the presence of BCL6 and PD1,
CXCR4, CXCR5, CXCL13, TBX21 and ICOS was found; as well as an inverse relationship between BCL6 and GATA3 expression. Interestingly, a set of microRNAs
(mir212, mi93, mir302b, mir520h, mir494, mir124a, mir10a, mir200b, mir181a*
and mir181c) predicted to control BCL6 expression were found statistically inversely
correlated to the presence of both BCL6 itself and many other TFH-related signature.
Conclusions: BCL6 expression in n-PTCLs is related to a TFH phenotype and AITL
morphology. Both BCL6 expression and the TFH phenotype in neoplastic T-cells
could be regulated by the same set of microRNAs that control the phenotype of
normal germinal center T-cells.
Keywords: BCL6, microRNAs, TFH
[PP-LYMP-026]
CLONALITY ASSAY IN EXTRANODAL NK/T CELL LYMPHOMA
Mineui Hong, So Young Kang, Younh Hyeh Ko
Sungkyunkwan University, Samsung Medical Center, Department of Pathology, Seoul, South Korea
Keywords: Diffuse Large B-cell Lymphoma, Molecular Subtyping
Background: Extranodal NK/T cell lymphoma (ENKTL), nasal type is designated
NK-cell and T-cell. According to WHO classification 2008, most cases appear to
be NK-cell neoplasm, nevertheless some cases are T-cell origin. The proportion of
T-cell origin of ENKTL and the type of T-cell has not yet been fully determined. The
present study was conducted to determine the proportion and the cell type of T-cell
origin ENKTL by using immunohistochemical study and T-cell receptor (TCR) gene
rearrangement.
Methods: We investigated 136 cases of ENKTL. 114 cases ENKTL originated from
nasal cavity and 22 patients developed extranasal sites. Immunohistochemical (IHC)
for F1 and TCRCM1 and TCR gene rearrangement following BIOMED-2 protocol
were performed. Repeat experiments with formalin-fixed paraffin-embedded tissue
for BIOMED-2 protocol were available in 120 cases.
Results: Based on TCR gene rearrangement by BIOMED-2, 33 cases (24.3%)
showed monoclonality. Among them, 17 cases (51.5%) expressed F1 or TCRCM1.
Remaining 16 cases (48.9%) were regarded as silent for T-cell expression. 103
| 17-22 October 2014
| EAHP - 2014
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89
Discussion: At least 39.7% of ENKTL were considered T-cell lineage. TCR gene
rearrangement is frequently silent in T-lineage ENKTL.  T-cell ENKTLs tend to
have better prognosis than NK-ENKTL or TCR-silent T-cell ENKTL. TCR-silent T-cell
ENKTLs which may include  T-cell ENKTL tend to have poorer prognosis.
Keywords: extranodal NK/T cell lymphoma, T-cell lineage, TCR gene rearrangement
Table 1. Overall survival of the patients according to NK/T-cell lineage
Characteristics of
patients
All patients NK-lineage
T-lineage p value
Median age
50.0(21-91)
49.5(21-86)
50.0(30-91)
Age (n=136)
0.57
50 years or less
72(52.9)
43(52.4)
29(53.7)
older than 50 years
64(47.1)
39(47.6)
25(45.3)
Male
44(32.3)
26(31.7)
18(33.3)
Female
92(67.3)
56(68.3)
36(66.7)
ECOG 0-1
81(85.3)
50(87.7)
31(81.6)
ECOG 2-4
14(14.7)
7(12.3)
7(18.4)
Limited, I-II
52(42.3)
32(44.4)
20(39.2)
Advanced, III-IV
71(57.7)
40(55.6)
31(60.8)
UNL or below
53(52.5)
30(52.6)
23(52.3)
over UNL
48(47.5)
27(47.4)
21(47.7)
Low/low intermediate
73(77.7)
41(77.4)
32(78.0)
High intermediate/high
21(22.3)
12(22.6)
9(22.0)
Negative
76(73.8)
44(74.6)
32(72.7)
Positive
27(26.2)
15(25.4)
12(27.3)
Negative
88(89.8)
49(89.1)
39(90.7)
Positive
10(10.2)
6(10.9)
4(9.3)
Negative
80(79.2)
47(81.0)
33(76.7)
Positive
21(20.3)
11(19.0)
10(23.3)
DETECTION OF MYD88 L265P IN VITREOUS BODY ASPIRATES
IMPROVES DIAGNOSIS OF INTRAOCULAR B-CELL LYMPHOMA
0.59
LDH (n=101)
1.00
B-symptom (n=103)
1.00
BM involvement (n=98)
1.00
LN involvement (n=101)
0.63
Clinical information was obtained from medial record and correlated with cell lineage of ENKTL.
αβ T-cell ENKTLs tend to have better prognosis than NK-ENKTL or TCR-silent T-cell ENKTL. (p=0.459.
[PP-LYMP-027]
SILENCER OF CYTOKINE SIGNALING 1 (SOCS1) GENE
MUTATIONS IN CLASSICAL HODGKIN’S LYMPHOMA
Jochen K. Lennerz1, Karl Hoffmann1, Nele Rüther1, Andreas Viardot2,
Peter Möller1
University Ulm, Institute of Pathology, Ulm, Germany
University Ulm, Department of Internal Medicine III, Ulm, Germany
Background: Gene mutations in the suppressor of cytokine signaling 1 (SOCS1) are
frequent in classical Hodgkin’s lymphoma; however, the prognostic relevance of
these mutations is unexplored.
Methods: We performed laser-capture microdissection of Hodgkin/Reed-Sternberg
(HRS) cells out of fresh-frozen (n=84) and formalin-fixed paraffin embedded (n=16)
tumor samples followed by full-length SOCS1 gene sequencing in a cohort of wellcharacterized patients with follow-up.
Results: SOCS1 mutations in HRS-cells are present in 61% of tumors
(n=52+9/84+16). Affected DNA-motifs as well as patterns of intratumoral accumulation imply that these mutations are the result of aberrant somatic hypermutation
| EAHP - 2014
| 17-22 October 2014
Irina Bonzheim1, Sabrina Giese1, Daniela Süsskind2,
Leticia Quintanilla Martinez1, Karl Ulrich Bartz Schmidt2,
Falko Fend1
1
2
1.00
IPI risk groups (n=94)
90
Keywords: SOCS1, prognostication, Hodgkin
[PP-LYMP-028]
0.56
Ann Arbor stage (n=123)
2
Conclusions: SOCS1 mutation status is a single-gene prognostic biomarker for over
60% of classical Hodgkin Lymphoma patients.
0.85
Performance status (n=95)
1
in HRS-cells. By predicted mutational consequence, the 61 mutated tumors can be
separated into those with HRS-cells that harbor non-foreshortening point mutations (‘minor’ n=47/61=77%) and tumors with HRS-cells that harbor at least one
foreshortening mutation (‘major’; n=14/61=22.9%). Clinical characteristics did not
differ between tumors with wild-type versus SOCS1 mutated, minor, or major HRS
cells, respectively. However, when outcome measures were compared to patients
with tumors composed of wild-type HRS-cells, SOCS1 mutations were associated
with favorable outcome. Importantly, subgroup analysis revealed that the prognostic difference was due to a significantly reduced relapse rate in classical Hodgkin’s
lymphomas with SOCS1 minor mutations in HRS-cells – whereas patients with HRScells that contained SOCS1 major mutations suffered from early relapse and demonstrated significantly shorter overall survival (P=0.03, log-rank). Thus, for nearly
two-thirds of the classical Hodgkin-Lymphoma patients, the SOCS1 mutation status
and in particular the mutation subtype predicts treatment course and overall survival.
1.00
Sex (n=136)
th
cases detected polyclonal or no peaks were categorized by IHC of F1 and
TCRCM1. Among 103 cases, 21 cases (20.4%) were positive for F1 or TCRCM1
and we assumed them to T-cell lineage. Classifying to NK-cell lineage and T-cell
lineage has no correlation with clinical findings.  T-cell ENKTLs tend to have better prognosis than NK-ENKTL or TCR-silent T-cell ENKTL and TCR-silent T cell ENKTL
tend to show worst, but it did not reach statistical significance.
Institute of Pathology, University Hospital Tübingen, Tübingen, Germany
Department of Ophthalmology, University Hospital Tübingen, Tübingen, Germany
Background: Intraocular lymphoma (IOL) is a rare malignancy. Most cases are diffuse large B-cell lymphomas and represent either primary IOL or involvement by
primary central nervous system lymphoma (PCNSL). B-NHL arising in immune-privileged sites such as the CNS show a high frequency of MYD88 mutations, up to 75%
in PCNSL. IOL is usually diagnosed by cytological, immunocytochemical and molecular examination of vitreous body aspirates. Separation from uveitis can be difficult,
and clonality analysis may be misleading due to pseudoclonal/oligoclonal patterns
resulting from low cellularity or benign B-cell clones in immunological disorders.
The aim of our study was to assess the frequency of MYD88 mutations in IOL and
to investigate their diagnostic potential in a large series of vitrectomy specimens.
Materials-Methods: DNA samples of 65 vitrectomy specimens of 60 patients with
a suspicion of IOL were retrospectively investigated for MYD88 mutations. Samples
had previously been examined by cytology and immunocytochemistry, and analysed
for immunoglobulin heavy chain (IGH) rearrangements by PCR. The MYD88L265P mutation was detected using an allele-specific PCR with melting point analysis. MYD88
exons 3 and 4 were sequenced in IOL samples wild type for MYD88L265P. Clinical
presentation, disease course, cytology and original diagnoses were reviewed and
correlated with the MYD88 mutation status.
Results: 28 males and 32 females with a median age of 73 years (range 23-94)
were included. A primary diagnosis of IOL was rendered in 16 samples/13 (22%)
patients, whereas a diagnosis of inflammation or insufficient evidence for IOL was
given in 49 samples/47 (78%) patients. Clonal IGH rearrangements were found in
10/13 IOL patients, whereas in 3 cases diagnosis was based on cytology and immunophenotype. MYD88L265P mutation was detected in 6/13 (46%) patients with
a diagnosis of IOL, and 7/13 patients showed MYD88WT. Among the 47 patients
diagnosed as negative or inconclusive for IOL, a MYD88L265P mutation was identified
in 7 samples/6 patients (13%), whereas the remaining 41 patients were MYD88WT.
A clonal IGH rearrangement was identified in 3/6 MYD88L265P patients without previous IOL diagnosis. Evaluation of clinical data and follow-up confirmed the diagnosis
of IOL in 18/60 (30%) patients including 12/13 with an initial IOL diagnosis and all
6 MYD88L265P patients originally considered negative or inconclusive for IOL. The
only patient with initial IOL diagnosis based on cytology and monoclonality by PCR
who was reclassified as viral retinitis revealed MYD88WT. A history of systemic or
CNS lymphoma was found in 5/18 (28%) IOL patients. In summary, MYD88L265P was
identified in 12/18 (67%) patients with confirmed IOL.
Conclusions: 1. Detection of MYD88 mutations is very useful for diagnosis of IOL in
the absence of cytological evidence of malignancy or clonal IG rearrangements. 2.
The high frequency of MYD88L265P in our series further supports that primary IOL and
PCNSL represent a single entity. 3. Clonal IGH rearrangements may occur in vitreous
body aspirates without evidence for IOL, indicative of benign B-cell clones
Keywords: Intraocular lymphoma (IOL), MYD88L265P
IMMUNOHISTOLOGICAL ANALYSIS OF THE JUN FAMILY
AND THE SIGNAL TRANSDUCERS AND ACTIVATORS OF
TRANSCRIPTION (STAT) IN THYMUS
ARRAY CGH BASED ANALYSIS OF POSTTRANSPLANT
PLASMACYTIC HYPERPLASIA REVEALS NO ABERRATIONS
Alexandra Papoudou Bai1, Alexandra Barbouti2, Anna Goussia1,
Vassiliki Galani2, Kalliopi Stefanaki3, Panagiotis Kanavaros2
Sylvia Hoeller, Thomas Menter, Darius Juskevicius,
Alexandar Tzankov
Institute of Pathology, University Hospital Basel, Basel, Switzerland
1
Department of Pathology, University of Ioannina, Ioannina, Greece
2
Departments of Anatomy-Histology-Embryology, University of Ioannina, Ioannina, Greece
3
Department of Pathology, Agia Sophia Childrens’ Hospital of Athens, Athens, Greece
The Jun family (c-Jun, JunB and JunD) and the signal transducers and activators
of transcription (STAT) proteins are involved in cell differentiation, proliferation and
apoptosis. Moreover, c-jun and STAT3 cooperate to regulate apoptosis. Therefore,
we analyzed by double immunostaining the immunotopographical distribution of
cells expressing Phospho-c-Jun (p-c-Jun), JunB, JunD, p-STAT3, p-STAT5 and pSTAT6 in thymus.
JunD was frequently expressed by thymocytes with higher expression in medullary
than cortical thymocytes. P-c-Jun, JunB and p-STAT3 were rarely expressed by
thymocytes whereas p-STAT5 and 6 were not detected in thymocytes.
P-c-Jun was frequently expressed by thymic epithelial cells (TEC) with higher expression in cortical than medullary TEC and Hassall Bodies (HB). The expression
levels of JunB and JunD were low in cortical TEC and higher in medullary TEC and
HB. P-STAT3 was frequently expressed by TEC with higher expression in cortical
than medullary TEC and HB. P-STAT5 and 6 were detected in rare TEC and HB.
Double immunostaining revealed p-c-Jun, JunB, JunD and p-STAT3 positivity in
cells expressing pan-cytokeratin (epithelial cells) or CD3 (T-cells) but not in cells
expressing markers of B-cells (CD20) or dendritic cells (S100 protein, CD123 or
CD207) or macrophages (CD163).
The diversity of the immunotopographical distibution and the expression levels of pc-Jun, JunB, JunD, p-STAT3, 5 and 6 in TEC provides immunohistological evidence
that their expression is tightly regulated and they are differentially involved in TEC
differentiation. Moreover, JunD may also play a role in thymocyte differentiation.
Keywords: PTLD, transplantation, nuclei enrichment
[PP-LYMP-031]
Keywords: Jun, STAT, thymus
Table 1. Expression of Phospho-c-Jun, JunB, JunD, p-STAT3, 5 and 6 in
thymic epithelial cells
Cortical
Medullary
Hassall bodies
+/++
+
+
JunB
-/+
+
+
JunD
+/++
+
+/++
p-STAT3
-/+
+/-
+/-
p-STAT5
-/+
-/+
-/+
p-STAT6
-/+
-/+
-/+
p-c-Jun
Post-transplant lymphoproliferative disorders (PTLD) are defined as lymphoid proliferations arising after solid organ or hematopoietic stem cell transplantation. Early
PTLD (ePTLD) are known to be polyclonal, but the absence of clonality does not
exclude malignancy and chromosomal abnormalities have been documented in
ePTLD of the follicular hyperplasia type. We analyzed genomic aberrations in plasmacytic hyperplasia (PH) at higher resolution. We selected five PH-ePTLD cases with
available formalin-fixed paraffin embedded (FFPE) material from the archive of our
institution. Applying a novel nuclear extraction technique we used a monoclonal antibody reactive against the nuclear protein multiple myeloma oncogene 1 (MUM1),
labeled plasma cell nuclei of PH and then isolated them by flow-sorting yielding
purity of >90%. Genomic DNA from the sorted plasma cell nuclei was isolated and
used for array-comparative genomic hybridization (aCGH) on the Agilent SurePrint
G3 CGH 180k arrays. We did not find any genetic copy number aberrations in our
cohort. Only two of the five cases showed in situ single detectable EBV-RNA positive
cells. However, all patients were EBV-seropositive at the time of transplantation.
After the diagnosis of ePTLD, immunosuppression was reduced in one of the five
patients, while in the others no adjustments of the immunosuppressive regimens
were done. None of the patients developed another form of PTLD or a relapse of
the ePTLD. Our findings might add momentum to the hypothesis that events other
than those occurring within PH-ePTLD are necessary to develop a neoplastic lymphoproliferation leading to more aggressive forms of PTLD. Thus, PH categorization
as PTLD might rather be reconsidered both due to the indolent clinical course and
based on these genetic findings showing lack of cytogenetic aberrations.
[PP-LYMP-030]
[PP-LYMP-029]
Julie Morscio1, Iwona Wlodarska2, Thomas Tousseyn1
1
2
Table 2. Expression of Phospho-c-Jun, JunB, JunD, p-STAT3, 5 and 6 in
thymocytes
Cortical
Medullary
p-c-Jun
-/+
-/+
JunB
-/+
-/+
JunD
-/+
+/++
p-STAT3
-/+
-/+
p-STAT5
-
-
p-STAT6
-
-
CLINICOPATHOLOGICAL STUDY OF THE ROLE OF THE EPSTEINBARR VIRUS IN 15 CASES OF RICHTER TRANSFORMED B-CELL
CHRONIC LYMPHOCYTIC LEUKEMIA (B-CLL)
KU Leuven, Department of Imaging and Pathology, Leuven, Belgium
Center of Human Genetics, KU Leuven, Leuven, Belgium
The Epstein-Barr virus (EBV) is an oncogenic human Herpesvirus that has been
associated with lymphomagenesis, in particular in immunocompromised individuals. Recently it has been suggested that in some cases EBV may play a role in the
transformation from the indolent B-cell Chronic Lymphocytic Leukemia (B-CLL) to
aggressive Diffuse Large B-cell Lymphoma (DLBCL), referred to as Richter transformation (RT), however little evidence supports this hypothesis.
The aim of this study is to retrospectively describe the clinicopathological characteristics in a Belgian unicenter study of 15 RT cases, with special attention for the
role of EBV in this process. We gathered the clinical characteristics from the patients
files, reviewed histopathology and performed a profound immunohistochemical
(IHC) screening of both the CLL and RT biopsies (including CD20, CD5, CD23, CD10,
BCL-6, MUM-1, BCL2, Mib1, C-MYC, CyclinD1, TP53, NOTCH1). All RT biopsies were
screened for the presence of EBV by means of EBV-encoded RNA (EBER) in situ
hybridization and EBV viral protein IHC. Immunoglobulin (Ig) PCR was performed on
the CLL and the RT to prove clonal relationship and determine the IgHV mutational
status, and karyotyping and fluorescent in situ hybridization (FISH) were performed
to determine the genetic profile.
In our series, there was a slight male predominance for RT. The median age at CLL
diagnosis was 67 years (range 38-76 years) and the median overall survival since
RT diagnosis was 4 months (range 1-33 months). EBER expression was detected
in 40% of RT DLBCL tested so far (analysis not complete at time of submission of
the abstract). The median interval between diagnosis of CLL and RT was 47 months
(range 16-106 months) and was shorter for EBV-related RT (median 33 months)
compared to EBV-unrelated RT (median 90 months). Although clonal relationship
was shown in all tested cases, the immunohistochemical profile showed some differences. RT DLBCLs showed a more complex karyotype than the CLL biopsies, and
| 17-22 October 2014
| EAHP - 2014
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91
th
the EBV-associated RT showed fewer genetic aberrations compared to the EBVunrelated RT.
In conclusion this small study suggests that there is a underestimated role for EBV
in the process of Richter transformation from CLL displaying different clinicopathological characteristics and underlying pathogenetic mechanisms that need further
investigation.
pattern of these genes defines the distinct latency programs (i.e. type I, II, and III latency). The latency I is characterized by the expression of only EBNA1 and EBER and
is reported in most of PBL and Burkitt lymphoma (BL). Since BL and PBL usually arise
in the setting of immunodeficiency, one should expect a latency III program; this may
suggest that the onset of these two diseases is not related to the imbalance of host
immunity but rather to other genetic mechanisms or to EBV itself.
[PP-LYMP-032]
In this study we aimed at investigating the role of EBV in the development of PBL
in comparison to BL, in terms of latency program expressed by the virus in the
neoplastic cells, and of microRNA (miRNA) profile to assess the contribution of viralencoded miRNAs in dysregulating host gene expression and affecting key cellular
pathways.
THE ROLE OF THE EPSTEIN-BARR VIRUS IN THE DEVELOPMENT
OF PLASMABLASTIC LYMPHOMA: NEW INSIGHTS INTO
CELLULAR AND VIRAL MICRORNA DYSREGULATION
EBV positivity was detected by EBER in situ hybridization and latency was assessed
by RT-qPCR, using Taqman probes and immunohistochemistry. MiRNA profiling (including both cellular and viral miRNAs) was performed using the MySeq Illumina
platform.
Giulia De Falco1, Maria Raffaella Ambrosio1, Lucia Mundo1, Sara Gazaneo1,
Satya V. Prasad Busarla2, Bruno Jim Rocca1, Vasileios Mourmouras1,
Stefano Lazzi1, Lorenzo Leoncini1
We found that, in addition to EBNA1, most of PBLs and BLs expressed LMP2 and
lytic genes (i.e. ZEBRA and EAD). LMP-1 was detected in one case of PBL and one
case of BL. These results suggest a non-canonical latency associated gene expression program with a subset of viral episome genes, corresponding to an abortive
lytic cycle. miRNA profile indicated that several EBV-miRNAs, belonging to the BART
family, are expressed both in EBV positive PBLs and in BLs, being expressed at a
higher level in PBL. Cellular miRNAs profile showed that only a few cellular miRNAs
are differential expressed between PBLs and BLs. In addition, when comparing BL
and PBL with the normal counterpart, principle component analysis showed that
PBLs and BLs clusterize together for cellular miRNAs and significantly differ from
any normal counterpart. If this our previous data will be confirmed, it would be
difficult to find for PBL and BL a normal cell to which they completely correspond.
Keywords: EBV, Richter transformation, B-CLL
1
Department of Medical Biotechnology, University of Siena, Italy
Moi University, Edoret, Kenya
2
The pathogenesis of Plasmablastic lymphoma (PBL) is not clearly understood, although a role for Epstein-Barr virus (EBV) has been proposed as EBV DNA is detectable in 60-75% of the cases. However, the exact mechanisms by which EBV may
contribute in promoting PBL is still an area of active debate. The proposed mechanism
for B-cell lymphomagenesis in the setting of EBV infection includes B-lymphocyte immortalization, leading to the emergence of dominant B-cell clones, and transcriptional
transactivation of viral and cellular genes resulting in B-cell transformation. It is well
known that EBV encodes a series of gene and microRNAs interacting with or exhibiting
homology to a wide variety of anti-apoptotic molecule, cytokines, and signal transducers. Transformed lymphocytes are more susceptible to other genetic mutations (i.e.
MYC gene rearrangement) that may enhance EBV lymphomagenesis. During latent
infection, to maintain the viral genome and to successfully evade the host cell immune surveillance, EBV expresses a small subset of genes. The differential expression
The reactivation of lytic cycle may play a role in EBV-driven lymphomagenesis by
increasing the total number of latently infected cells. The detection of LMP2 reflects
the molecular environment that MYC needs to unfold its oncogenic potential. In addition, EBV-miRNAs may promote the transformation of primary B lymphocytes by
dysregulating physiologic pathways.
Keywords: EBV, microRNA, plasmablastic lymphoma
Figure 1. EBV positive vs. plasmablastic.
92
| EAHP - 2014
| 17-22 October 2014
IGH-FR1 ANALYSIS BY MASSIVE PARALLEL SEQUENCING
FOR THE DETECTION OF CLONALITY AND SOMATIC
HYPERMUTATIONS IN LYMPHOID MALIGNANCIES
Anna Gazzola1, Ying Huang Huang2, Jeff Panganiban2,
Michael Klass2, Jeffrey Miller2, Claudia Mannu1, Stefano Aldo Pileri1,
Pier Paolo Piccaluga1
1
Hematopathology Unit, Department of Experimental, Diagnostic, and Specialty Medicine, Bologna
University, Bologna, Italy
2
Research and Development, Invivoscribe Technologies, Inc., San Diego, California, USA
The standard method to analyze clonality and somatic hypermutations (SHM) of immunoglobulin genes (IG) in B-non Hodgkin lymphomas (B-NHLs) is a multiplex PCR
followed by capillary electrophoresis and/or Sanger sequencing. This technique is
fast and fairly accurate; however it is subject to inherent limitations and clonality
assessment remains partially subjective.
Recently, massive-parallel sequencing (MPS) of immune receptor genes was demonstrated to be highly effective, with increased sensitivity for detecting sequences of interest.
Based on that, it was proposed for the diagnosis and monitoring of lymphoid neoplasms.
We designed a phase 3 diagnostic accuracy study aiming to compare the value of the
first commercially available kit for MPS of IGH/FR1 with the gold standard analysis based
on BIOMED2 reactions. We analyzed 64 samples, including 58 B-NHL and 6 reactive
lymphoid hyperplasia (RLH), obtained from peripheral blood (PB) (N=46) or formalinfixed paraffin-embedded tissue (FFPE) (N=18). IGH rearrangements were evaluated with
both traditional capillary electrophoresis and MPS. First, each sample was subjected to
multiplex PCR amplification of IGH repertoire according to EuroClonality guidelines followed, in clonal samples, by Sanger sequencing of FR1 region in order to establish the
mutational status of IGH. Second MPS was performed with LymphoTrack IGH FR1 assay
and IGH Leader assay by Invivoscribe Technologies on an Illumina MiSeq.
[PP-LYMP-033]
IGH alignment and mutational status was analyzed on IMGT (the international
ImMunoGeneTics information system http://www.imgt.org). Diagnostic accuracy evaluation was performed by CATmaker software (Centre for Evidence Based Medicine,
Oxford University).
Conventional clonality analysis indicated 55 clonal and 8 polyclonal cases. One case
was not evaluable due to amplification failure and was then excluded. MPS identified
53 clonal and 10 polyclonal cases. The overall diagnostic accuracy was 96.4% (53/55
cases), ST, SP, PPV, and NPV being 96%, 100%, 100%, and 80%, respectively (Figure1).
The two discrepant cases referred to highly degraded samples.
Mutational spectrum of IGH was performed in 46 clonal samples by conventional
and NGS analysis. Conventional mutational analysis showed 12 samples with germline IGH, 8 with a mutation burden ranging from 1.1 to 3.0%, and 19 with mutated
IGH, while 7 samples were not evaluable. Conversely, MPS demonstrated 15 samples with germline IGH, 5 with a mutation burden ranging from 1.0 to 3.0%, 26 with
mutated IGH. The two MPS assays (IGH FR1- and Leader-assay) showed a perfect
concordance (R2=0.99, p<0.0001). Similarly, conventional detection and MPS gave
very comparable results (R2=0.71; p<0.0001) (Figure 2). Notably, MPS allowed the
analysis of all cases, including the 7 that failed by Sanger sequencing.
In conclusion, MPS appeared very effective for IGH analysis in lymphoid disease
(even on FFPE samples) and possibly superior to conventional tools concerning
SHM detection. In order to apply them to routine diagnostic, MPS based approaches
should now be evaluated prospectively and cost-effectiveness assessment should
be performed.
Keywords: MPS, clonality, B-NHL
Figure 2. Comparison of somatic mutation detection with conventional vs MPS
methods. A) regression plot of FR1 vs Leader analysis by MPS; B) regression plot of
Sanger vs MPS.
[PP-LYMP-034]
TUMOR SUPPRESSOR MIRNAS 29C AND 146A ARE
SPECIFICALLY ASSOCIATED WITH THE ALK+ ALCL SIGNATURE
AND REVEAL POTENTIAL INVOLVEMENT IN TUMOR CELL
DISSEMINATION
Julia Steinhilber, Ann Kathrin Gersmann, Kathrin Dieter, Falko Fend,
Leticia Quintanilla Martinez, Irina Bonzheim
Institute of Pathology and Neuropathology, University Hospital Tübingen, Eberhard-Karls-University,
Tübingen, Germany
Background: ALK+ anaplastic large cell lymphoma (ALCL) is a systemic disease
affecting lymph nodes and extranodal sites. We recently investigated the differential
expression of miRNAs between ALK+ ALCL, ALK- ALCL and normal T-cells using
next generation sequencing (NGS). ALK+ ALCL showed a specific miRNA expression
pattern compared to ALK- ALCL or T cells. The aim of this study was to characterize
the function of two top miRNA candidates associated with the signature of ALK+
ALCLs; miRNAs 29c and 146a, which are expressed at very low levels in ALK+ ALCL
when compared to T-cell and ALK- ALCL. In order to identify their target genes, a
transcriptome analysis was performed and validated.
Figure 1. Diagnostic accuracy of IVS NGS assay. Diagnostic accuracy of IVS NGS
assay (Clonality detection FR1) - CATmaker software (Centre for Evidence Based
Medicine, Oxford University).
Design: The ALK+ ALCL cell line SUDHL-1 treated with transfection reagent or
transfected with miRNA mimics 29c or 146a (GE Healthcare) were analyzed by
transcriptome analysis perfoming next generation sequencing. Some of the identified regulated target genes of miRNA 29c or 146a were validated by RT-qPCR in
SUDHL-1 and Karpas 299 cell lines. Additionally, miRNAs 29c- or 146a-dependent
| 17-22 October 2014
| EAHP - 2014
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93
Results: Transcriptome analysis resulted in 153 and 18 target genes significantly
regulated by miR-29c and miR-146a, respectively. RT-qPCR validation of most
strongly regulated and functionally most interesting genes revealed nine downregulated candidate genes - CCNF, COL6A3, DOT1L, TDG, RCC2, ADAM19, GGCT,
CCNA2 and LMNB1 - after overexpression of miR-29c mimic in SUDHL-1 and
Karpas 299 cells. Overexpression of miR-146a in SUDHL-1 and Karpas 299 cells
resulted in down-regulation of four genes - ZNF275, SRPRB, PNPO and BSG - in
transcriptome and RT-qPCR analysis. All of these genes are predicted direct targets
of miR-146a or miR-29c by the miRanda tool possessing bindings sites in their 3’UTRs. Gene reporter assay showed direct regulation by the miRNAs 29c or 146a for
all but one (BSG) candidates. All nine identified miR-29c target genes are described
as highly expressed in various tumors and have important functions in tumorigenesis, such as in angiogenesis, differentiation, proliferation and cell cycle. Both miR146a target genes SRPRB and BSG are reported to have oncogenic functions and to
be overexpressed in solid tumors. Interestingly, a common function of several of the
miR-146a and 29c targets (BSG, ADAM19, LMNB1 and COL6A3) is to drive cell dissemination and invasion in solid tumors, indicating a potential role of these miRNAs
in migration and invasion of ALK+ ALCL cells.
Conclusions: 1) miRNAs 29c and 146a are specifically associated with the ALK+
ALCL signature and are consistently downregulated compared to normal T-cells and
ALK- ALCL. 2) The oncogenic target genes identified show high expression in ALK+
ALCL. BSG, ADAM19, LMNB1 and COL6A3 are mainly involved in cell dissemination
and invasion and might be responsible for the peculiar dissemination and invasion
pattern known for ALK+ ALCL in lymph nodes. 3) Our data provide new insight into
the biology of ALK+ ALCL.
Keywords: ALK+ ALCL, miRNAs
th
expression was validated at protein level by Western blot analysis. Direct regulation
of the target genes by miRNAs 29c or 146a was analyzed using a gene reporter
assay.
[PP-LYMP-035]
C-MYC EXPRESSION AND MYC ALTERATION IN MANTLE CELL
LYMPHOMA
Ji Young Choe1, Ji Yun Yun1, Jooryung Huh2, Soojin Shin2,
Hyun Jung Kim3, Jin Ho Paik1, Young A. Kim4, Yoon Kyoung Jeon5,
Ji Eun Kim4
1
Department of Pathology, Seoul National University Bundang Hospital, Seongnam, Korea
Department of Pathology, University of Ulsan College of Medicine, Asan Medical Center, Seoul,
Korea
3
Department of Pathology, Inje University Sanggye Paik Hospital, Seoul, Korea
4
Department of Pathology, Seoul National University Boramae Hospital, Seoul, Korea
5
Department of Pathology, Seoul National University Hospital, Seoul, Korea
2
Background: Mantle cell lymphoma (MCL) is a rare type of B-cell lymphoma characterized by histologically indolent but clinically aggressive features. Recently, overexpression of c-myc has been found to be one of the important prognostic factors in
a subset of high grade B-cell lymphomas aside from Burkitt lymphoma. We aimed
to investigate c-myc expression and MYC alteration in mantle cell lymphoma and to
evaluate their clinical significance.
Methods: A total of 64 patients with MCL including 54 conventional, 8 blastoid and
2 pleomorphic variants were enrolled in this study. Expression of c-myc was tested
by immunohistochemistry (IHC) and quantitatively assessed using an automated
image analysis system. Gene amplification or translocation of MYC was examined
by fluorescent in situ hybridization (FISH).
Results: Overall, expression of c-myc was very low, showing negativity in the
majority of cases (mean positivity: 2.7%, median: 1.02%). However, blastoid variant of MCLs showed higher expression of c-myc (mean: 7.9%) than conventional
types (mean: 1.90%, p=<0.001). Amplification of MYC was found in one case of 50
tested (2%), which was a blastoid variant, presenting the highest c-myc expression
(29.7%) by IHC. Translocation of MYC was not found (0/50). There was no significant
correlation or association between c-myc status and various clinicopathologic parameters including patients’ outcome.
Conclusions: Alteration or activation of c-myc might be related to the pathogenesis
of MCL, blastoid variant. Further studies should be allotted to find MYC alteration
other than translocation or amplification.
[PP-LYMP-036]
NON-HODGKIN B-CELL LYMPHOMA WITH PROLYMPHOCYTIC
MORPHOLOGY (NH-BLPM): RELEVANCE OF AN INTEGRATIVE
DIAGNOSTIC APPROACH
Roberto Hernández Mora1, Jorge Garcia Vera1,
Fernando Perez Jacobo2, Lorena Viramontes Aguilar1,
Jose Tovar Bobadilla1, Ricardo Aguilar Guadarrama1,
Leticia Quintanilla Fend3, Carmen Lome Maldonado1
1
Pathology department Instituto Nacional de Ciencias Medicas y Nutricion “Salvador Zubiran”
Mexico city, Mexico
2
Hematology department Instituto Nacional de Ciencias Medicas y Nutricion “Salvador Zubiran”
Mexico city, Mexico
3
Institute of Pathology, University of Tüebingen. Tüebingen Germany
Introduction: B-cell Lymphoproliferative neoplasms with prolymphocytic morphology represents a heterogeneous group of neoplasms poorly studied. Despite sharing
the same morphology, sometimes, they exhibit different clinical behavior and genetic abnormalities. Recognized lymphoproliferative disorders with this morphology
are B-cell chronic lymphocytic leukemia/ small lymphocytic lymphoma (CLL / SLL),
B-cell prolymphocytic leukemia (LPL- B) and mantle cell lymphoma. We report 3
cases of NH-BLPM, unusual clinical features and genetic alterations involving c-Myc
and CCND-1 genes, proving the importance of an integrative diagnostic approach of
this peculiar group of entities.
Purpose of the study. To present the clinical, morphological, immunophenotypical
and genetic features in 3 cases of NH-BLPM diagnosed between January of 2002
and December of 2013.
Materials-Methods: We reviewed 3 cases of B-cell lymphomas with prolymphocytic
morphology from the pathology archives of our institution. Morphologic analysis
of the peripheral blood smear, lymph node and bone marrow aspirate and biopsy
with additional immunohistochemical studies on the biopsies were performed.
Cytogenetic analysis of the bone marrow aspirate, FISH for t(11;14) and break
apart probe for c-Myc, PCR for IGH, PCR for t(11;14) and RT-PCR for Cyclin D1 was
performed.
Results: See Table 1.
Discussion. In accordance with other observations, the spectrum of genetic alterations in prolymphocytic proliferations is variable. Our study allows us to determine
the genetic features with diagnostic relevance that leads us to 2 different molecular
patterns.
The first pattern (case 1) is the association between prolymphocytic morphology
with genetic alterations in the CCND1 gene. These cases are considered blastoid
variants of mantle cell lymphoma, regardless their morphology or Cyclin D1 immunohistochemical expression. The t(11;14) has been reported in many non Hodgkin
B-cell lymphomas and seems to be associated with aggressive behavior. However,
the biologic significance in this particular group of neoplasms is not clear.
The second molecular pattern (cases 2 and 3) includes neoplasms with prolymphocytic morphology and genetic anomalies on the c-Myc gene. Most of these cases
(case 2) showed lymphocytosis with more than 55% of prolymphocytes in peripheral blood and bone marrow; these clinical data coupled with the immunophenotype
and the presence of translocation with rearrangement of c-Myc, these cases are
considerated B-PLL.
Case 3 presented significant nodal involvement, aggressive course, bulky disease,
prolymphocytic morphology and evidence of Burkitt-type translocation t(8;14),
which did not meet the WHO-2008 B-cell prolymphocytic leukemia diagnostic criteria. The clinical and biological significance of this entity is unclear.
Conclusions: NH-BLPM are a heterogeneous group of neoplasms. They are associated with genetic alterations involving the CCDN1 and c-Myc genes. They can present lymph node involvement with or without a leukemic phase. We propose the term
“non-Hodgkin B-cell lymphoma with prolymphocytic morphology” as a morphologic
diagnosis to those neoplasms that does not fulfill the diagnostic criteria for other
lymphoproliferative disorders. We emphasize the relevance of clinical, morphological, immunohistochemical and genetic correlation for the diagnostic approach of
this group of neoplasms.
Keywords: prolymphocytic, CCND-1, c-Myc
Keywords: c-myc; mantle cell lymphoma; in situ hybridization, fluorescence
94
| EAHP - 2014
| 17-22 October 2014
Case2
Case3
50/Female
51/Male
56/Male
WBC count/L
3.6 x10^9
16x10^9
11x10^9
8%
60%
5%
Prolymphocytes in
bone marrow
Splenomegaly
Absent
Absent
Absent
Lymphadenopathy
Present
Present
Present
Chemotherapy
Yes
Yes
Yes
Bone Marrow
Transplantation
Yes
No
No
Follow up
10 years, WD
7 years, WD
10 months, PR
CD20+, CD5-, IgM+,
CD23-, CD43-,
BCL6-, KI67 40%
CD20+, CD5-, IgM+,
CD23-, CD43-,
BCL6+, KI67 5%
CD20+, CD5+,
IgM+, CD23-, CD43BCL6-, KI67 80%
Cyclin D1
Negative
Negative
Negative
P27
Immunophenotype
Negative
Positive
Positive
PCR Cyclin D1
NP
Negative
Negative
PCR t(11;14)
NP
NP
Negative
t(11;14) in 40% of
cells
t(2;8)
t(8;14)Myc/IGH in
20% of cells
Clonal
rearrangement
Clonal
rearrangement
Clonal
rearrangement
NP
t(2;8)(p12;q24)
NP
FISH
PCR IGH
Cytogenetics
NP: Not performed, WD: Without disease, PR: Partial remission
Conclusion: Our findings confirmed the high incidence of BRAF V600E in patients
with suspected cHCL and the absence of the aberration in other B-cell lymphomas.
Molecular analysis could complement the current diagnostic armamentarium, particularly using DNA from bone marrow samples in patients without recognizable
leukemic cells. However, the mutation detection in patients with features of HCLv
and phenotype switch towards cHCL, as well as the absence of the marker in some
cHCL cases, reported also by other authors, raises the question of disease heterogeneity. Further studies are needed to test the novel diagnostic possibilities at the
edge of the target treatment era.
Ackowledgements: The study was partially supported by the National science Fund.
Keywords: hairy cell leukemia, flow cytometry, BRAF V660E
[PP-LYMP-038]
FOLLICULAR LYMPHOMAS WITH SINGLE INGUINAL NODAL
PRESENTATION ARE FREQUENTLY BCL2 NEGATIVE: A
CHALLENGING DIAGNOSIS
Elena Sabattini1, Riccardo Panzacchi1, Teresa Marafioti2,
Claudio Agostinelli1, Simona Righi1, Carlo Sagramoso Sacchetti1,
Georgia Levidou1, Francesca Rosini1, Stefano Pileri1
1
2
[PP-LYMP-037]
MOLECULAR DETECTION OF BRAF V660E MUTATION IS
COMPLEMENTARY TO MORPHOLOGY AND FLOWCYTOMETRY
IN HAIRY CELL LEUKEMIA
Margarita L. Guenova1, Tihomir I. Dikov1, Antoaneta S. Michova1,
Vasil G. Hrischev2, Gueorgui N. Balatzenko1
1
Specialised Haematology Laboratory Block, National Specialised Hospital for Active Treatment of
Haematological Diseases, Sofia, Bulgaria
Haematology Clinic, National Specialised Hospital for Active Treatment of Haematological
Diseases, Sofia, Bulgaria
2
Background: Hairy cell leukaemia (HCL) is an indolent mature B-cell neoplasm,
characterized by specific morphological and immunophenotypic criteria of tumour
cells involving peripheral blood and diffusely infiltrating the bone marrow and
splenic red pulp. Making the correct diagnosis is essential for applying effective
treatment with purine analogues resulting in durable remissions and an overall
10-years survival rate exceeding 90%. However, diagnostic difficulties can often
arise because of a low number of circulating leukemic cells and a “dry tap” on
marrow aspiration. Since 2011, when by using whole exome sequencing, Tiacci et
al. identified a V600E mutation in exon 15 of the BRAF gene in HCL, great interest
has been caused in the opportunity to incorporate the detection of the mutation into
diagnostic testing.
Aim: To explore the feasibility and diagnostic implication of BRAF V600E mutation
identified by PCR in patients with hairy cell leukemia.
Materials-Methods: We investigated 10 patients with classical hairy cell leukemia,
as well as 2 patients with HCL variant and a control group of patients with other
B-cell lymphomas. Peripheral blood was evaluated for the presence of recognizable hairy cells and a bone marrow biopsy with immunohistochemistry and reticulin
staining was performed. Flow cytometry on peripheral blood or bone marrow samples was done using an 8-colour panel of monoclonal antibodies, FACS Canto II and
Diva software ver.6.1.2. For BRAF V600E mutation detection, DNA was isolated from
peripheral blood and/or bone marrow aspirate and/or deparaffinized bone marrow
biopsy. Allele-specific PCR was performed.
Results: Patients with classical HCL (cHCL) were characterized by the presence of
cytopenia and splenomegaly. In 9/12 patients hairy cells were recognized in the peripheral blood, including patients with HCLv, while in 3 – the lymphoid cells showed
insignificant mature morphology. Flow cytometry, based on the bright expression of
CD20, CD22 and CD11c, CD103 and CD25 allowed for the detection of 1-61% of
Unit of Haemolymphopathology, S.Orsola, Malpighi Hospital, University of Bologna, Italy
University College London, Department of Pathology, United Kingdom, Pathology
Case1
Age/ Gender
hairy cells in the examined cHCL samples. HCLv lacked CD25, however, in one of
the cases the marker appeared two years after diagnosis. PCR analysis allowed for
the detection of BRAF V600E mutation in 9 out of 10 patients with cHCL, including
patients in whom the detection of less than 0.02 G/l cells with HCL phenotype challenged the diagnosis, as well as in the patient previously diagnosed with HCLv who
at the moment of the molecular analysis showed an acquired positivity of CD25.
The second HCLv patient and other B-cell lymphoma patients were found negative.
Table 1. Summary of clinical, immunohistochemical and genetic features
Introduction: Follicular lymphomas (FLs) arising at inguinal lymph nodes often show
peculiar morphology for the physiological hyaline-fibrotic features of such organs
and the lytic appearance of the follicles, which may challenge the diagnosis. These
cases have been frequently found positive for CD23 but no data on BCL2 expression
have been specifically reported, which may be crucial for correct interpretation.
Aims: To assess whether FLs primarily arising at the inguinal level (stage IA) are
characterized by peculiar immune-phenotype with special reference to BCL2
expression.
Materials-Methods: Thirty-nine FLs were collected over the period 2007-2014, all
presenting as stage IA with single inguinal nodal involvement. FLs grade IIIB or
with predominantly diffuse growth or areas of transformation into a diffuse large
B-cell lymphoma were excluded. All cases were immunohistochemically studied
for CD3, CD20, BCL6, CD10, Ki-67/MIB1 and BCL2 (clones 124 and E17); 10 of 13
BCL2 negative cases were also studied with the anti-BCL2 clone SP66 and 33/39
samples were stained for IRF4. FISH analysis for t(14;18) was carried out in all BCL2
negative cases.
Results: Of 39 patients, 21 were females (53.8%) and 18 were males. The median age was 50 years (range 26-82, average 49) and 11/39 cases (28.2%) were
younger than 40 years most of whom (9/11: 81.81% ) showing a grade 3a FL. On
the whole, grades 3a were 24/39 (61.53%).
13/39 (33.3%) were BCL2 negative with the anti-BCL2 antibodies (clones 124 and
E17) as were the 10 cases stained with anti-BCL2 SP66 clone. No significant difference was observed between BCL2 staining and age: positive cases occurred at a
median age of 48 (range 26-62, average 46), while negative ones at a median age
of 52 (range 37-82, average 54). No significant difference in grade 3a cases was
recorded between the 2 groups. All BCL2 negative cases lacked t(14;18). BCL6 was
always positive in >75% of tumour cells, as well as CD10, which was expressed in
all biopsies but one (a BCL2 positive case). IRF4 was expressed only in a minority
of grade 3a cases (15.8%).
Conclusions: FLs primarily presenting at inguinal site seem to have distinctive features since they 1) occur in a younger population than non-inguinal FLs (median 50
years), with slightly less than 30% presenting in patients younger than 40 years who
mostly show grade 3a histology, 2) display BCL2 negativity both on immunohistochemistry and FISH in more than 30% of cases. The latter feature must be carefully
considered to reduce possible misdiagnoses. The reason of this site-related higher
ratio of BCL2 negative cases and younger age at onset are unknown, but to some
extent resemble what already known for childhood and testicular FLs.
Keywords: Follicular lymphomas, BCL2, inguinal lymph node
| 17-22 October 2014
| EAHP - 2014
|
95
th
[PP-LYMP-039]
DIFFUSE LARGE B CELL LYMPHOMA DERIVED FROM NODULAR
LYMPHOCYTE PREDOMINANT HODGKIN LYMPHOMA PRESENTS
WITH VARIABLE HISTOPATHOLOGY
Sylvia Hartmann1, Mine Eray2, Claudia Döring1, Tuula Lehtinen3,
Uta Brunnberg4, Paula Kujala2, Martine Vornanen2,
Martin Leo Hansmann1
1
Dr. Senckenberg Institute of Pathology, Hospital of the Goethe University, Frankfurt am Main,
Germany
2
Department of Pathology, Tampere University Hospital and University of Tampere, Tampere 33520,
Finland
3
Department of Oncology, Tampere University Hospital, Tampere 33520, Finland
4
Department of Internal Medicine 2, Hospital of the J. W. Goethe University, Frankfurt am Main,
Germany
lymphoma (DLBCL). Immunophenotypically, 97% of cases (69/71) were of B cell
origin of which all but 1 were EBV positive. Donor derived PTLD was predominant
(91%, 65/71); 6 (9%) were of host origin. Among 5 umbilical cord blood (UCB) stem
cell transplant recipients, one of them was also transplanted with T cell depleted
peripheral blood stem cell (TCD PBSC). This patient developed cord blood derived
MALT lymphoma, EBV negative. Patients with multi-organ involvement had a higher
mortality rate (87.5%) compared to those with single-organ involvement (15.8%)
(P<0.001).
Conclusions: PTLD is a heterogenous disease with a wide spectrum of clinical, morphologic, and molecular genetic features ranging from reactive polyclonal lesions to
frank lymphomas. While the predominent PTLD post bone marrow transplant are
EBV driven and donor derived, up to 10% of cases are of host origin and 4% are
EBV negative.
Keywords: allogeneic hematopoetic, EBV post transplant lymphoproliferative disorders, engraftment studies
Table 1. Clinical and pathological features of 71 PTLD cases
Nodular lymphocyte predominant Hodgkin lymphoma (NLPHL) usually presents in
middle aged men and shows an indolent clinical behavior. However, up to 30% of
the patients present a secondary transformation into aggressive diffuse large B cell
lymphoma (DLBCL). The aim of the present study was to characterize morphology
and immunophenotype of this kind of DLBCL in detail and compare it with conventional DLBCL.
Morphology and immunophenotype of 33 cases of NLPHL with simultaneous or sequential transformation into DLBCL were investigated. These cases were compared
with 41 de novo DLBCL in Finnish men.
Sex
Male:46
Female:25
Age at
diagnosis
(years)
Median:
31
Range:
5-73
EBV status
Pos: 68
(95.8%)
Neg: 3
(4.2%)
Unmodified
(BM/PBSC): 7
TCD PBSC:
59
UCB
only: 4
UCB+TCD
PBSC:1
DLBCL:
56 (78.9)
DLBCL/CHL:
1 (1.4%)
MALT:
1 (1.4%)
T-NHL:
2 (2.8%)
GI/respiratory: Multi-organs:
16 (22.5%)
8 (11.3%)
Brain:
2 (2.8%)
Skin/soft
tissue:
2 (2.8%)
TCR only:
2 (2.8%)
NA:
31 (43.7%)
Type of
transplantation
The majority of composite lymphomas exhibited different immunophenotypes in the
NLPHL and the DLBCL components. The immunophenotype of the DLBCL secondary
to NLPHL was heterogeneous. However, BCL6, EMA, CD75 and J-chain were usually
expressed in both components (>= 73% positive). Overall, the NLPHL component
was more frequently positive for EMA, CD75 and J-chain than the DLBCL component. In contrast, B cell markers, CD10 and BCL2, were more frequently expressed
and were expressed at higher levels in the DLBCL component than in the NLPHL
component. In the independent series of de novo DLBCL 4 cases could be identified
with a growth pattern and immunophenotype that suggested that they had arisen
secondarily from NLPHL.
Pathology
P-PTLD:
11 (15.5%)
Site of
involvement
LN/Tonsil:
43 (60.6%)
The morphology and immunophenotype of DLBCL arisen from NLPHL is heterogeneous. Further characterization of the particular molecular features of this subgroup
is warranted.
Origin of PTLD
Donor:
65 (91%)
Host:
6 (9%)
IgH/TCR
rearrangement
IgH only
(DLBCL):
31 (43.7%)
IgH+TCR+:
5 (7%)
Onset of PTLD
Median:
4 mos
Range:
2 mos-5 yrs
Follow-up
(months)
Keywords: Nodular lymphocyte predominant Hodgkin lymphoma, diffuse large B
cell lymphoma, transformation
Last follow up
status
IgH-TCR-:
2 (2.8%)
Range: 0-211
Death from
PTLD: 7
Death from
others: 10
Alive:
54
[PP-LYMP-040]
POST TRANSPLANT LYMPHOPROLIFERATIVE DISORDER:
CLINICOPATHOLOGIC SPECTRUM BASED ON A SINGLE
INSTITUTION EXPERIENCE
Jinjuan Yao, Justyna Sadowska, Ahmet Dogan, Maria E. Arcila
Department of Pathology, Memorial Sloan Kettering Cancer Center, New York, NY. 10065 USA
Background: Post transplant lymphoproliferative disorder (PTLD) is a complication
of bone marrow transplantation with a high mortality rate. Clinical, morphologic and
molecular genetic features are heterogeneous and the incidence varies depending
on the patient risk and type of transplant. Published single institution experience
remains limited by a relatively small number of patients. In this study we describe
the clinicopathologic and molecular features of 71 PTLD cases after allogeneic hematopoietic stem cell transplantation from a single institution.
Methods: Seventy one (71) cases of PTLD diagnosed at Memorial Sloan Kettering
Cancer Center between 1993 and 2014 were identified from the Department of
Pathology files. Clinical and pathologic data were collected. All available morphologic, immunophenotypic and engraftment studies were reviewed.
Results: The clinical and pathological features are summarized in Table 1. Patients
were predominantly male (65%, 46/71) with a median age of 31 years (range 5-73
yrs). Most cases represented single site involvement (88.7%, 63/71) with lymph
node and tonsils as predominant sites. Eight patients (11.3%, 8/71) presented with
multiorgan involvement, two with concurrent CNS disease. Onset of PTLD after
transplantation was at a median of 4 months (range 2 mos - 5 yrs). Morphologically,
the majority were monomorphic PTLD (84.5%, 60/71), primarily diffuse large B-cell
96
| EAHP - 2014
| 17-22 October 2014
[PP-LYMP-041]
PEDIATRIC PRIMARY CUTANEOUS MARGINAL ZONE
LYMPHOMA
Marianne Tinguely1, Werner Kempf1, Dmitry V. Kazakov2,
Stanislaw A. Buechner3, Mario Graf4, Andreas Zettel5,
Dieter R. Zimmermann6
1
Kempf and Pfaltz, Histologische Diagnostik, Zurich, Switzerland
Department of Pathology, Faculty of Medicine in Pilsen, Charles University in Prague, Czech Republic
3
Histologische Diagnostik, Basel, Switzerland
4
Dermatology Practice, Zurich, Switzerland
5
Viollier Pathology Laboratory, Basel, Switzerland
6
Molecular Diagnostics, Institute for Surgical Pathology, University Hospital Zurich, Switzerland
2
Background: Low-grade B-cell lymphomas (lgBCL) are exceedingly rare in the pediatric population in general. The same is true for primary cutaneous marginal zone
lymphoma (PCMZL), which however seems to represent the most common lgBCL in
this age category. The differential diagnosis of PCMZL includes especially Borreliaburgdorferi-induced pseudolymphoma cutis which can be difficult to distinguish
due to morphological overlaps. Although PCMZL has an excellent 5-year OS, no
standardized therapy regimens are recommended so far.
Aim: To sum up biological and clinical presentation of three pediatric PCMZL and to
compare them with adult ones
Result: The patients were 13-18 years of age, one female and two male individuals with a follow up time of 9-41 months. All presented with multiple nodules. All
Keywords: pediatric, PCMZL
Figure 1. Detected viruses across the samples in eBL. 6 of known viruses have
been detected. The presence of Herpes virus 4 (EBV) in 100% of the samples is a
positive control. 7/20 cases present HHV5 (cytomegalovirus), 5/20 cases present
HHV8 (KSHV), 1/20 case presents HTLV1.
[PP-LYMP-042]
IS BURKITT LYMPHOMA A POLYMICROBIAL DISEASE?
EVIDENCE FROM RNA SEQUENCING
Francesco Abate1, Giulia De Falco2, Maria Raffaella Ambrosio2,
Maria Antonella Laginestra3, Lucia Mundo2, Sara Gazaneo2,
Fabio Fuligni3, Cristiana Bellan2, Pier Paolo Piccaluga3,
Stefano Aldo Pileri3, Raul Rabadan1, Lorenzo Leoncini2
1
Department of Biomedical Informatics, Center for Computational Biology and Bioinformatics
Columbia University, New York, USA
Department of Medical Biotechnology, University of Siena, Italy
3
Department of Specialistic, Diagnostic and Experimental Medicine, University of Bologna, Italy
2
Recent data provides insights into the emerging concepts of polymicrobial disease
pathogenesis for endemic Burkitt lymphoma (eBL). In particular, the potential mechanisms, by which Plasmodium falciparum and Epstein-Barr virus (EBV) infection
could interact have been addressed. Plasmodium falciparum impacts on immunity
and viral persistence by exhaustion of EBV specific T-cell response and Toll-like
receptor (TLR)-9 mediated reactivation of EBV latently infected memory B-cells.
EBV encodes several latent proteins essential for viral immortalization of B cells,
but EBNA1 protein is the only EBV latent protein consistently expressed in eBL.
Other EBV latent and lytic transcripts are not consistently expressed, although they
have been occasionally reported in a subset of eBL tumors. Therefore the underlying
mechanism linking EBV infection of B-cells to the emergence of malignancy remains
undiscovered.
Figure 2. Analysis of EBV in eBL. Q1: What is the latency of EBV in eBL? Q2:
Is the virus reactivated or latent (lytic or lysogenic phase)? Our findings revealed a
diversity of non-canonical latency associated gene expression program with subset
of viral episomes initiating lytic reactivation.
Conclusion: PCMZL rarely occurs in pediatric patients and should be distinguished
from pseudolymphoma. However, their biology and clinical presentation overlaps
with adult ones.
showed a light chain restriction and 2/3 had a clonally rearranged IgH by PCR. No
association with either IgG4 or Borrelia-burgdorferi (serology and PCR) was found.
The treatment included excision in all three patients and additionally antibiotics or
intralesional INFalpha injection in one of each.
In addition, epidemiological studies suggest that malaria and EBV alone cannot account for the eBL cases in high risk regions. In fact, malaria and EBV are ubiquitous
within the lymphoma belt of Africa and, unless other cofactors are involved, the
tumour should be much more common than it is. Arboviruses and plant tumor promoters are other possible local cofactor.
To determine if previously unidentified pathogens are present in BL and to understand their contribution in the pathogenesis of this disease, we established a tissue
sample cohort that could be assayed for viral RNA.
We used a subtractive algorithmic approach by which the input reads were aligned
to host reference with different alignment program (bowtie2, blastN and Megablast)
and a computational subtraction was applied at each step. Resulting unmapped nothost reads were the input for pathogens identification. 6 of known herpes viruses
have been identified in tumor samples. The presence of Herpes virus 4 (EBV) in
100% of the samples was a positive control. 7/20 cases presented Human herpes
virus (HHV) 5 (cytomegalovirus), 5/20 cases presented HHV8 (Kaposi Sarcomaassociated herpes virus - KSHV), 1/20 case presented Human T-lymphotropic virus
(HTLV) 1. These results were validated by PCR and immunohistochemistry on primary tumors which confirmed the presence of these additional pathogens in the
tumors microenvironment. We then deeply investigated the latency of EBV in eBL
and asked the question whether the virus could be reactivated (lytic or lysogenic
phase). Our findings revealed a diversity of non-canonical latency associated gene
expression program with a subset of viral episomes initiating lytic reactivation as
indicated by expression of genes corresponding to lytic program. These results were
validated by RT-qPCR and immunohistochemistry in primary tumors samples.
Our data supports the view that BL is a polymicrobial disease and that lytic EBV
reactivation may also contribute to the development of EBV-associated lymphomas.
Taken together, these findings suggest potential therapeutic strategies for virusassociated cancer.
Keywords: Burkitt lymphoma, infectious agents, RNA sequencing
Figure 3. EBV signature in eBL.
| 17-22 October 2014
| EAHP - 2014
|
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th
[PP-LYMP-043]
[PP-LYMP-044]
LYMPHOMATOID GRANULOMATOSIS AS A MODEL OF EBV
ASSOCIATED LYMPHOMAS
B LYMPHOBLASTIC LEUKEMIA/LYMPHOMA ARISING IN
PATIENTS WITH FOLLICULAR LYMPHOMA IS DISTINCT FROM DE
NOVO B ALL AND IS ASSOCIATED WITH A POOR PROGNOSIS
Cristiana Bellan1, Teresa Amato1, Stefano Lazzi1,
Santiago Montes Moreno2, Lorenzo Leoncini1
1
Anatomical Pathology Section, Department of Medical Biotechnology, University of Siena, vie delle
Scotte, 6, 53100, Siena, Italy
Hospital Universitario Marques de Valdecilla, Santander, Spain
2
Lymphomatoid granulomatosis (LYG), is a rare extranodal Epstein-Barr virus (EBV)associated B-cell lymphoroliferative disorder (LPD). Pathologically, LYG is an extranodal angiocentric and/or angiodestructive B-cell lymphoproliferative disorder
composed of polymorphic lymphoid infiltrate that consists of T lymphocytes, plasma
cells, and atypical larger neoplastic EBV(+) B-cells. Therefore, EBV lies at the heart
of LYG and is intrinsically linked to all aspects of its pathogenesis and pathophysiology. In healthy carriers, following primary infection, the EBV typically remains in a
latent state within memory B cells during which time it is regulated and controlled
by cytotoxic T cells (CTLs)—both CD8+ and CD4+—and natural killer (NK) cells.
Failure of the host’s cellular immunity can lead to EBV-induced B-cell proliferation
and when this occurs, infected carrier B-cells can transform from their latent state
into malignant cells and this can lead to the development of lymphoma.
In most cases of LYG and of other LPDs the disease arise in patients without overt
immunodeficiency. Moreover, a role for T cells in LYG has been inferred from the
presence of large T cell infiltrates that are constantly associated with this LPD.
The Aim of this project is to clarify the role of EBV in the development of LYG, first
of all to assess the type of latency in LYG that is still unclear, by sequence analysis
of rearranged immunoglobulin VH region genes; second to elucidate the possible
role of T cell component in this LPD, by analyzing T-cell antigen receptor (TCR)
repertoire.
Amy Rich1, Carlos E. Bueso-Ramos1, Sherry A. Pierce2, Susan O’Brien2,
Hagop M. Kantarjian2, Jeffrey Medeiros1, Sergej Konoplev1
1
Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, USA
Department of Leukemia, The University of Texas MD Anderson Cancer Center, Houston, USA
2
Background: Patients with follicular lymphoma (FL) may subsequently develop
TdT-positive blastic neoplasms that resemble and are often classified as B acute
lymphoblastic leukemia (B-ALL). In this study, we collected a small series of these
rare tumors to better understand their clinicopathologic and prognostic features.
Design: We searched our institutional archives for patients with the diagnosis of
B-ALL who also had a documented history of FL. Clinical and diagnostic data were
obtained by review of medical records. The comparison group included patients of
similar age with Ph-negative B ALL without a history of any malignancies.
Results: Between 1999 and 2013 708 patients with a diagnosis of B-ALL presented
to our institution for treatment. Eleven patients (1.5%) had a history of FL. Patient
6 first developed TdT-negative double-hit B-cell lymphoma and then TdT+ blastic
neoplasm. We also assessed a control group of 123 patients with Ph-negative B
ALL age 45-70. The study group demonstrated significantly worse overall survival.
Conclusion: TdT-positive blastic neoplasms arising in patients with a history of FL
have a very poor prognosis. We suggest that the term B-ALL is best avoided for
these rare tumors as they are likely distinct from cases of de novo B-ALL.
Keywords: B-ALL, TdT, progression
The improve understanding of the biology of this lymphomas including elucidating the role that EBV plays in his pathogenesis could paved the way for improved
therapies targeted at critical signaling pathways as well as the development of novel
cellular therapies in EBV associated lymphomas.
Keywords: Lymphomatoid granulomatosis, EBV, Immunoglobulin rearrangement
Figure 1. Overal survival. Unfavorable overall survival of patients with
transformed B-ALL compared to patients with de novo B-ALL
Table 1. Clinical and laboratory characteristics of patients with B-ALL and
preceding follicular lymphoma
Figure 1.
Table 1. VDJ usage in LYG
Age/
sex
47 M
FL
grade
2
Interval,
mo
9
58 M
3A
88
61 W
3A
53
62 W
2
82
60 M
1
4
Phenotype
CD10+/CD20+/TdT+/
sIgk+
CD10+/CD20+/
CD34+/TdT+/sIgCD10+/CD20+/
CD34+/TdT+/sIgCD10+/CD20+/
CD34-/HLADR+/
TdT+/sIgCD10+/CD20+/
CD34-/HLADR+/
TdT+/sIgCD10+/CD20+/
CD34-/TdT+/sIg-
number of
cases
Range of
mutation
Presence of ongoing
mutations
59 W
3A
25
M/UM
9
9/9
89.24-96.86
yes
67 M
1
11
CD10+/CD20-/
CD34-/TdT+/sIg-
62 M
1
13
CD10+/CD20-/
CD34-/TdT+/sIg-
39 M
2
11
CD10+/CD20+/
CD34-/TdT+/sIgCD10+/CD20+/
CD34-/TdT+/sIgCD10+/CD20+/
CD34+/TdT+/sIg
Results of immunoglobulin gene rearragemente analysis. M: mutated; UM: unmutated
54 M
2
9
64 M
3A
10
Cytogenetics
ND
ND
Followup, mo
5
46,XY
ND
126
Alive
46,XX,del(20)
(q11.2)
Complex with
t(8;14)
ND
15
Dead
MYC
11
Dead
Complex with
18+
ND
4
Dead
ND
MYC,
BCL2/
IgH
MYC,
BCL2/
IgH
ND
1
Dead
5
Dead
3
Dead
ND
6
Dead
ND
11
Dead
ND
7
Dead
Complex with
t(8;14), t
(14;18)
46,XY,del(11)
(q23)
[2]/46,XY[18]
ND
Complex with
18+
ND
FISH
Status
Dead
Age, at the time of lymphoma diagnosis; M, man, W, woman, FL, follicular lymphoma, Interval, time between FL and B ALL
diagnosis; mo, months; ND, not done
98
| EAHP - 2014
| 17-22 October 2014
Milhan Telatar, Hao Hong, Carrie Louie, Vandana Sharma,
Cindy Fong, Dennis Weisenburger, John Chan, Patricia Aoun
Department of Pathology, City of Hope National Medical Center, Duarte, USA
Hematological malignancies are characterized by the accumulation of somaticallyacquired alterations in genes that have diagnostic, therapeutic and prognostic impact. Testing the mutational status of each clinically-actionable gene individually by
conventional platforms such as Sanger sequencing is labor-intensive, requires a
large amount of DNA and results in prolonged turn-around-times. As large scale sequencing efforts are broadening the catalog of mutations in hematological cancers,
it is imperative to establish high-throughput testing with comprehensive profiling in
the clinical laboratory setting. We successfully overcame these limitations in solid
tumor testing by validating the 50-gene AmpliSeq™ Cancer Hotspot Panel from Life
Technologies on the Ion Torrent Personal Genome Machine (PGM) (Life Technologies)
for solid tumors. A custom designed 100-gene panel is now being validated on
the Illumina MiSeq platform for clinical testing of hematological malignancies. The
100-gene panel was designed using Agilent SureDesign software with most stringent and maximized performance probe tiling parameters and 100% coverage of all
exons of each gene. Screening for mutations in these genes will provide important
information for prognosis, risk stratification and appropriate management of patients with hematological malignancies. Validation of the test panel is in progress.
Keywords: Post-Rituximab, pure plasmacytic differentiation, relapsing
Table 1. Histopathological and immunohistochemical results
Case
1
Keywords: Multi-gene mutational profiling, next generation sequencing, hematological malignancies
[PP-LYMP-046]
PURE PLASMACYTIC INFILTRATION OF BONE MARROW
AND EXTRAMEDULLARY SITES OF PATIENTS WITH NODAL
OR EXTRANODAL B-CELL LYMPHOMA RELAPSING POSTRITUXIMAB
George Kanellis1, Aliki Xochelli2, Asimina Papanikolaou1,
Evdoxia Pouliou1, Garifalia Kokkini3, Ekaterini Stefanoudaki4,
Paraskevi Roussou5, Konstantinos Papanastasiou6,
Niki Stavroyanni2, Achilles Anagnostopoulos2,
Konstantinos Stamatopoulos7, Theodora Papadaki1
Initial
Diagnosis1
Post Rituximab1
Tissue
Diagnosis
Tissue/Percent
infiltration/
clonality (PCD)
Stomach
MALT with clonal Α(κ) (PCD)
BM/8%/Α(κ)
BM/ 5%/Μ(κ)
2
Stomach
MALT with clonal Μ(κ) (PCD)
3
Lymph node
LPL Μ(κ)
BM/7%/Μ(κ)
4
Lymph node
Nodal MZL with clonal G(κ) (PCD)
BM/25%/G(κ)
5
Stomach
BM/ 15%/Μ(κ)
6
Abdominal mass
MZL with clonal Μ(κ) (PCD); or LPL
BM/ 35%/Μ(κ)
7
eye
BM/ 10%/Μ(κ)
8
eye
MALT with clonal Μ(λ) (PCD)
BM/ 10%/Μ(λ)
9
Lung
BALT with clonal Μ(λ) (PCD)
BM/ 4%/Μ(λ)
10
Stomach
MALT with clonal (κ) (PCD)
Stomach / - / (κ)
11
Stomach
MALT with clonal G(κ) (PCD)
Stomach/ - /G(κ)
12
Epiglottis
MALT with clonal ΜD(κ) (PCD)
Epiglottis / - /MD(κ)
13
Lymph node
DLBCL,NOS with clonal G(λ) (PCD)
BM/2%/G(λ)
14
BM
MZL with clonal Μ(κ) (PCD); or LPL
BM/ 15%/Μ(κ)
15
BM
MZL with clonal Μ(κ) (PCD)
BM/ 5%/Μ(κ)
16
BM
MZL
BM/ 3%/Μ(κ)
MULTI-GENE MUTATIONAL PROFILING OF HEMATOLOGICAL
MALIGNANCIES BY NEXT GENERATION SEQUENCING IN A
CLIA-LICENSED LABORATORY
Conclusions: Our study confirms and significantly extends previous findings that
B-cell lymphomas with/without PCD at initial diagnosis can relapse after Rituximabcontaining therapy with pure plasmacytic infiltration in the absence of B-cell infiltration confirmed by negativity for multiple B-cell markers (CD20, CD79, PAX5). As
already suggested (Mahevas et al. J Clin Invest 2013), a possible undelying mechanism revolves around the induced differentiation of long-lived plasma cells that is
promoted by the milieu generated by B-cell depletion. For diagnostic purposes,
pathologists need to be aware of this rare phenomenon and investigate follow-up
biopsies post-Rituximab with a broad panel of immunohistochemical markers in
order to differentiate induced pure plasmacytic infiltration clonally related to the
original tumor from a distinct plasma cell dyscrasia with different clonality from that
of the initial lymphoma.
[PP-LYMP-045]
In all cases lymphomatoid cells had the following immunophenotype: CD20+, CD79α+, PAX5+, bcl6-,
CD10-, Cyclin D1-, CD5-, CD23-, CD3-.
1
1
Hematopathology Department, Evangelismos Hospital, Athens, Greece
Hematology Department and HCT unit, G. Papanicolaou Hospital, Thessaloniki, Greece
3
Hematology Department, Sismanoglio Hospital, Athens, Greece
4
Hematology Department, Agioi Anargyri Hospital, Athens, Greece
5
3rd University Clinic of Internal Medicine, Sotiria Hospital, Athens, Greece
6
Hematology Department, Agios Savvas Hospital, Athens, Greece
7
Institute of Applied Biosciences, Center for Technology and Research Hellas, Thessaloniki, Greece
2
Introduction: Anti-CD20 induced B-cell depletion is a widely used therapeutic strategy for a broad range of B-cell disorders. Most relevant experience stems from the
extensive use of Rituximab for the treatment of B-cell lymphomas as well as various
autoimmune diseases. In both contexts, rarely, disease recurrence post-Rituximab
manifests as pure plasmacytic population in the absence of the B-cell component.
Few such cases of B-cell lymphomas have been reported in the literature.
Aim: We here describe our experience from the retrospective analysis of 16 cases of
B-cell lymphomas with/without plasmacytic differentiation (PCD) at the initial diagnosis who relapsed after treatment with Rituximab-containing regimens with pure
plasmacytic infiltration of the bone marrow and/or extramedullary sites.
Patients and Methods: The study included 16 patients (7 males, 9 females), aged
52-78 years. The following biopsy samples were evaluated for the initial diagnosis
of B-cell lymphoma: stomach (n=5), lymph nodes (n=3), bone marrow (BM) (n=3),
eye (n=2), epiglottis (n=1), abdominal mass (n=1) and lung (n=1). The biopsy
samples evaluated at disease relapse post-Rituximab were as follows: BM (n=13),
stomach (n=2) and epiglottis (n=1). All samples were evaluated morphologically
(H&E) and immunohistochemically with the following markers CD20, CD79, PAX5,
CD3, CD5, CD23, CyclInD1, MUM-1, bcl6, CD10, CD138, SIg/CIg (, , , μ, , ).
Results: Histopathological and immunohistochemical results are summarized in
Table 1.
[PP-LYMP-047]
CONCOMITANT BCL2 AND BCL6 GENE REARRANGEMENTS IN
GERMINAL CENTER-DERIVED B-CELL LYMPHOMAS: INDOLENT
“DOUBLE HITS”?
Bettina Bisig1, Anna Talamo1, Chloé Boéchat1, Carmen Barcena1,
Christiane Copie Bergman2, Laurence de Leval1
1
Institute of Pathology, Centre Hospitalier Universitaire Vaudois, CH-1011 Lausanne, Switzerland
Department of Pathology, Assistance Publique - Hôpitaux de Paris (APHP), Groupe hospitalier
Henri Mondor - Albert Chenevier, F-94010 Créteil, France
2
Background: BCL2 or BCL6 gene rearrangements are detected in a variable fraction
of follicular lymphomas (FL) and diffuse large B-cell lymphomas (DLBCL). “Doublehit” B-cell lymphomas, strictly defined as bearing concomitant breaks in MYC plus
another oncogene, are clinically aggressive and show frequent extranodal involvement. Dual BCL2 and BCL6 gene rearrangements have drawn less attention in the
literature.
Materials-Methods: To characterize the pathological and clinical features of B-cell
lymphomas with concomitant BCL2 and BCL6 gene rearrangements we retrieved
from our files all B-cell lymphomas tested by FISH with break-apart probes for BCL2
and BCL6 (Dako), and for high grade cases also for MYC (Abbott Molecular). The
literature on dually BCL2/BCL6 translocated B-cell lymphomas was also reviewed.
Results: We identified 9 cases with dual BCL2 and BCL6 translocations in the
absence of MYC rearrangement. Mean age at presentation was 65.3 years,
male:female ratio was 4:5. Concomitant BCL2 and BCL6 rearrangements were
| 17-22 October 2014
| EAHP - 2014
|
99
th
documented at initial diagnosis in 7 cases (4 grade 1-2 FL, one grade 3a FL, 2
DBLCL with a germinal center-like immunophenotype). One case (grade 1-2 FL with
BCL2 break) acquired BCL6 gene aberration at relapse, without significant variation
of the histology. In the last patient the initial biopsy had shown a grade 1-2 FL but
could not be analyzed by FISH; a subsequent biopsy (9 years later) revealed similar
histology and dual BCL2/BCL6 rearrangements. Seven cases had a BCL2+/CD10+/
BCL6+ immunophenotype, while 2 were BCL2+/CD10-/BCL6+ (one FL and one
DLBCL). The lesions were extranodal in 6 cases (67%). Affected sites included oral
(2 cases), gastric or colonic mucosa (one case each); uterus (one case); and skin
(one case). Bone marrow involvement was present at diagnosis in 4 of 7 informative cases (57%). Two patients with long follow-up presented with nodal grade 1-2
FL; the first patient showed cutaneous progression 17 years later; the second one
relapsed with bone marrow involvement 4.5 years after diagnosis.
Table 1 provides a summary of the clinical and pathological findings of our case
series and 30 other such cases (28 FL and 2 DLBCL) previously reported in the
literature. Cases with FL histology were more commonly grade 1-2, except in a
series from China where all cases were grade 3. Follow-up was available for a few
patients. In the series by Keller et al., one patient with FL grade 3a died 1 year after
diagnosis; 2 patients with FL grade 1-2 showed no evidence of progression 1.3
and 1.5 years after diagnosis; 2 patients with DLBCL were alive 3.6 and 7 years
after diagnosis. Daudignon et al. reported that one patient (FL) died 0.8 years after
diagnosis; another patient (FL) transformed into an aggressive lymphoma 2.3 years
after diagnosis; 2 patients (FL) were in remission 0.7 and 1.3 years after diagnosis.
Conclusion: Concomitant BCL2 and BCL6 gene rearrangements occur in B-cell
lymphomas of germinal center derivation (FL and DLBCL). In this small series they
most commonly presented with extranodal involvement and showed dual BCL2/
BCL6 translocations already at initial diagnosis. In contrast to double-hit lymphomas
with MYC locus aberrations, these BCL2/BCL6 dually rearranged B-cell lymphomas
appeared to have a relatively indolent clinical behavior, although follow-up data
were limited and differences might exist between different regions of the world.
Keywords: BCL2, BCL6, rearrangement
Extranodal involvement (%)
Bone marrow involvement (%)
2 (2/0)
7/9
(78%)
6/9
(67%)
4/7
(57%)
9
65.3
Pan Y et
al., Diagn
Mol Pathol
2012;21:234
(Asia)
8
NA
NA
8 (0/8)
0
NA
NA
NA
Gollub W et al.,
Anticancer Res
2009;29:4649
(Western
Europe)
10
NA
NA
10
(7/3)
0
9/10
(90%)
NA
5/10
(50%)
Keller CE et al.,
Am J Clin Pathol
2008;130:193
(North America)
8
2 (1/1)
7/8
(88%)
0/8
(0%)
1/3
(33%)
Daudignon A
et al., Cancer
Genet Cytogenet
1999;111:157
(Western
Europe)
4
Sex
Initial diagnosis
Mean age in years (range)
(GC/nGC)
4M/5F 7 (6/1)
Number of patients
DLBCL
Bisig B et al.,
current
(Western
Europe)
Reference
(provenance)
100
FL
(G1-2/G3)
IHC CD10+ (%)
Table 1. Summary of main clinical and pathological features of reported
dually BCL2/BCL6 rearranged B-cell lymphomas
(47-89)
68.7
1M/3F 6 (5/1)
(32-88)
| EAHP - 2014
56.3
0M/4F
4
0
NA
(39-68)
NA
NA
[PP-LYMP-048]
CYTOGENETIC AND FLOW CYTOMETRY EVALUATION
OF RICHTER SYNDROME REVEALS MYC, CDKN2A, IGH
ALTERATIONS WITH LOSS OF CD52, CD62L AND INCREASE
OF CD71 ANTIGEN EXPRESSION AS THE MOST FREQUENT
RECURRENT ABNORMALITIES
Grzegorz Rymkiewicz1, Renata Woroniecka2, Katarzyna Blachnio1,
Beata Grygalewicz2, Jolanta Rygier2, Zbigniew Bystydzienski1,
Beata Sledz Gawronska1, Kinga Sikorska Mali1,
Barbara Pienkowska Grela2, Monika Prochorec Sobieszek3
1
Flow Cytometry Laboratory, Pathology Department and Laboratory Diagnostics, The Maria
Skłodowska-Curie Memorial Institute and Cancer Centre, Warsaw, Poland
2
Cancer Genetics Laboratory, Pathology Department and Laboratory Diagnostics, The Maria
Skłodowska-Curie Memorial Institute and Cancer Centre, Warsaw, Poland
3
Pathology Department and Laboratory Diagnostics,The Maria Skłodowska-Curie Memorial Institute
and Cancer Centre, Warsaw, Poland; Institute of Hematology and Transfusion Medicine, Warsaw,
Poland
Background: Richter syndrome (RS) is a transformation of chronic lymphocytic
leukemia (CLL) or small lymphocytic lymphoma (SLL) into high-grade lymphoma,
especially diffuse large B-cell lymphoma, not otherwise specified (DLBCL,NOS).
Diagnosis of RS is most commonly based on histopathological examination or occasionally on flow cytometry (FCM) of cell suspensions/peripheral blood with a constellation of clinical features. Comparison of the FCM of the previous CLL/SLL has
revealed minor modulations of antigen expression without major alterations in most
of RS cases.
Methods: In this study, FCM, classical cytogenetics (CC), fluorescence in situ hybridization (FISH) and cytology smear (CYT) were performed in 8 cases diagnosed
as RS. Clinical presentation included estimation of lymph nodes status, LDH level,
treatment and outcome of CLL/SLL and RS. RS and CLL/SLL were evaluated by
numerous antibodies. Antigen expression was categorized into three groups: [-] no
expression, [+/-] expression on >20%<100% of cells, and [+] expression on 100%
of cells. Expression of CD (19, 20, 22, 23, 52, 79), FMC7, HLADR, and CD (5, 25, 38,
43, 45, 52, 62L), ZAP-70, BCL2 on DLBCL,NOS cells was described as mean fluorescence intensity (MFI) value in comparison to MFI of these antigens on CLL/SLL
and T-lymphocytes, respectively. This procedure allows to estimate whether a given
antigen has a higher [+] or weaker [+] expression and it enables to compare the
expression of pan-B antigens with each other. Lymphoma subtype was defined according to WHO 2008 classification. CC studies were done on cells obtained by FNAB
or PB. Interphase FISH was performed on the fixed cells using probes as follows: IGH
BAP, TP53, ATM, D13S319/13q34, CEP12, MYC BAP, CDKN2A/CEP9, IGH/BCL2, IGH/
MYC/CEP8 and IGH/CCND1.
Results: Characteristics of patients are given in Table 1. Five pts were previously diagnosed as CLL and 2 as SLL. In 1 patient diagnosis of de novo RS was made without a history of CLL/SLL. The CYT and FCM results are summarized in Table 2 and
exemplary cases shown in Figure 1, 2. RS cells were larger than CLL/SLL cells, both
in CYT and FSC/SSC scattergram. All RS cases were CD45, CD19, CD43, HLADR,
BCL2, CD38, ZAP-70, CD71 positive. Proliferative activity measured by CD71 was
detected in all pts on 100% of RS cells. In all RS cases,  expression of CD38 and
ZAP-70 was found. The MFI of CD19 expression was  compared with  MFI of
CD20 in 7 out of 8 cases. The 7 RS cases had immunophenotypic features similar
to CLL/SLL. Comparison of immunophenotypes revealed recurrent lost or decreased
expression of CD62L (all cases) and CD52 (7 of 8 cases), increased expression of
CD71 (all cases) and lost or decreased (7 cases) IgD expression in RS cells. CC
analysis was successful in 7 out of 8 RS cases (Table 3). In all cases the karyotypes
were complex. FISH ( 7 of 8 cases) showed MYC alterations in all pts. Homozygous
or heterozygous CDKN2A deletion was detected in 5 of 7 cases. Four of 7 cases
showed IGH@ rearrangement.
Conclusions: The majority of RS were characterized by a loss/decrease of CD52,
CD62L and increased CD71 expression. CC identified complex karyotypes with
losses of 9/9p and 17/17p as the most frequent. All RS cases demonstrated MYC
abnormalities. Disruptions of CDKN2A and IGH were identified in most cases. In addition, loss of CD52 expression found in our study, most likely predicts for resistance
to the alemtuzumab therapy frequently used in CLL.
Keywords: Richter Syndrom, Flow cytometry, Genetics
| 17-22 October 2014
Figure 2. FNAB/FCM analysis of the case no. 4. Dot-plots showing the
coexistence of normal T lymphocytes and SLL cells (red cells in R1 region) with
larger DLBCL, NOS cells (green cells in R2 region). A) FSC/SSC scattergram
showing two distinct clusters of cells: small T lymphocytes and SLL cells (R1, red)
and larger DLBCL, NOS cells (R2, green). B) CD19 vs. CD5, SLL and DLBCL,NOS
showing the same level of CD19 but CD5 coexpression is weaker compared with
normal T lymphocytes. C) CD71 vs. CD62L, DLBCL, NOS has increased intensity
of CD71 and decreased CD62L expression compared with the SLL. D) CD79β vs.
CD20, DLBCL, NOS has both decreased CD79β and CD20 expression compared
with the SLL. E) CD22 vs. CD23, DLBCL, NOS has increased intensity of CD23 and
loss of CD22 expression compared with the SLL. F) CD19 vs. CD52, DLBCL,NOS
has decreased and lost CD52 expression compared with the SLL.
| 17-22 October 2014
67/M
51/M
55/M
4
5
6
DLBCL, NOS (EP)
DLBCL, NOS (LN, BM)
DLBCL, NOS (LNm, S, BM)
DLBCL, NOS (LNm)
DLBCL, NOS (PB, BM, S,
H, CNS)
Transformed CLL/SLL
(localization)
CLL (PB, BM, LN, C)
CLL (PB, BM)
DLBCL, NOS (C)
DLBCL, NOS (PB)
CLL/SLL (LNm, PB, BM) DLBCL, NOS (PB, BM)
SLL (EP)
CLL (PB, LN)
-
CLL (PB, BM, LN)
SLL (LN)
Initial CLL/SLL
(localization)
yes b
no
no
no
yes a
yes a
yes a
yes a
Rapidly asymmetric generalized
lymphadenopathy a/Tumors b
yes
yes
yes
no
yes
yes
yes
yes
LDH>UNV
15
88
24
48
52
0
66
12
Time to RS (mo)
DOD(0)
DHAP (NR); CN3OP (PR); B; DOD (9)
RB (NR); ICE (NR); DOD (3)
R-CHOP (PR); 1x CODOX-M/IVAC (PR/progression); OFAR
(PR/ progression); RTG/R; GMALL; DOD (4)
CHOP (NR); VAD (PR); DOD (1)
DX +MTX (PR); VAD (PR); AraC+ Dx +MTX; DOD (3)
Treatment after Transformation (response)
and outcome (mo)
R-FC (CR)
R-CHOP + DepoCyte (CR), AWED (6)
L (PR); COP (PR); CHOP (CR); CC (CR); CHOP;DOD (1)
FC (PR)
2CdA,
+CTX (CR)
R-CHOP (NR); R-FC+ LP +RTG (CR)
FC (PR); R-CHOP (PR)
-
LP (PR); R-FC (CR)
R-CHOP (PR)
Treatment before transformation
(response)
| EAHP - 2014
|
AWED -Alive, with evidence of disease; AraC+Dx+MTX – arabinozide, cytarabine, dexamethasone, methotrexate; B – bendamustine; BM - bone marrow; C - cutaneous and subcutaneous bulky tumors; CC - cyclophosphamide, cladribine; 2CdA+CTX – cladribine+cyclophosphamide; CHOP – cyclophosphamide, adriamycin, vincristine, prednisolone; CLL - chronic lymphocytic leukemia;
CN3OP - mitoxantrone, cyclophosphamide, vincristine, prednisone;CNS - central nervous system; CODOX-M/IVAC - cyclophosphamide, vincristine, doxorubicin, methotrexate, ifosfamide, etoposide, arabinozide cytarabine; COP - cyclophosphamide, vincristine, prednisolone; CR - complete remission; DHAP - dexamethasone, arabinozide, cytarabine, cisplatin; DLBCL, NOS - diffuse
large B-cell lymphoma, not otherwise specified; DOD - deed of disease (progression); Dx+MTX - dexamethasone, methotrexate; EP - extranodal presentation; F-female; FC - fludarabine, cyclophosphamide; GMALL (GMALL - German Multicenter Adult ALL Study Group) - the alternate use of drugs (rituximab, fractionated cyclophosphamide or ifosfamide, vincristine, methotrexate,
cytarabine, teniposide and prednisone or doxorubicin); H- hepatomegaly; ICE - ifosfamide, carboplatin, etoposide; L- leukeran; LDH>UNV, Increased lactate dehydrogenase above the normal reference value; LN - lymph node; LNm - massive lymphadenopathy; No – case number; LP - leukeran, prednisolone; NR - no response; M – male; mo – months; OFAR - oxaliplatin, fludarabine,
cytarabine, rituximab; PB - peripheral blood; PR - partial remission; R- rituximab; RB - prednisone, bendamustine; R-CHOP – rituximab, cyclophosphamide, adriamycin, vincristine, prednisolone; R-FC - rituximab, fludarabine, cyclophosphamide; RTG – radiotherapy; S – splenomegaly; SLL - small lymphocytic lymphoma; VAD - vincristine, doxorubicin, dexamethasone.
61/M
28/M
3
8
61/F
2
79/F
64/F
1
7
Age/Sex
No.
Table 1. Basic clinical data of patients with Richter syndrome
Figure 1. Morphologic Features of Richter Syndrome. A) Fine needle aspiration
biopsy of lymph node (case 2) demonstrating a mixture proliferation of large
cells with round nuclei, prominent, often centrally located eosinophilic nucleolus
(immunoblast morphology), and cells with two to four mostly marginal nucleoli
(centroblast morphology) (H&E, original magnification 400). B) Peripheral blood
smear of DLBCL,NOS (case 6) demonstrating 2 morphologically distinct cell
populations. One prominent population consists of larger cells with prominent
nuclear irregularities, less condensed chromatin, variable nucleoli, and moderate
amounts of basophilic cytoplasm. The second small population is composed of small
round lymphocytes of CLL with scant cytoplasm and condensed chromatin and
normal T lymphocytes (Wright-Giemsa, original magnification 1000). C) Peripheral
blood smear of DLBCL,NOS (case 7) demonstrating 2 morphologically distinct cell
populations. One prominent population consists of huge cells compared than in
case 6 with prominent nuclear irregularities, less condensed chromatin, variable
nucleoli, and moderate amounts of basophilic cytoplasm. The second population is
composed of small round CLL lymphocytes (Wright-Giemsa, original magnification
1000).
101
102
| EAHP - 2014
| 17-22 October 2014
↑
↑
↑
↑
↑
↑
3
4
5
6
7
8
IB
LB
LB
IB
IB
IB
IB/CB
IB
↑
↑
↑
↑
↑
↑
↑
↑
+
+
+
+
+
+
-
+
+↓
+↓
+↑
+↓
+↓
+
+
+↓
+
+
+↑
+↓
+
+
+
+
CD45 CD19
+dim
+bright
+bright
+/-dim
+/-dim↓
+/-dim
-↓
+dim
CD20
+
+
-
+
+
+
+
+
CD19>CD20
-
+
-
+
+
+
+
+
CD5
-↓
+/-
-
+↑
+↑
+
+/-
+
+↑
+
+↓
+
+↑
+
+
+
CD23 CD43
+↑
ND
+↑
+↑
+↑
+
+↓
+
HLADR
+↑
+↑
+↑
+↑
ND
+↑
+↑
+↑
+↑
+↑
+↑
+↑
+↑
+↑
+↑
+↑
+↑
+↑
ND
+↑
+
+
+↑
+↑
-
+/-↑
-
+/-
+
+
+
-
+
-
-
+↑
+
-
+
+
+
-
+↑
-
+/-↓
-
-
-
+↑
-↓
ND
ND
+/-↓
+/-
-↓
+/-
-
+/-
+
-
-
-
-
-
-
-
+
-
-
-
-
-
+↓
-↓
-↓
-↓
+/-↓
-↓
-↓
+/-↓
+↓
-↓
+↑
-↓
-↓
+/-↓
+/-↓
-↓
+++↑
+++↑
+++↑
+++↑
+++↑
+++↑
+++↑
+++↑
λ↑
-
κ
λ
-
λ
λ↓
-
CD38 ZAP-70 BCL2 CD11c CD25 CD22 CD79B FMC7 CD10 CD62L CD52 CD71 κ / λ
D↓, M
M, G
M
-
D↓, M
M
D↓, M
M
sIg
ND
ND
ND
46,XY[21]
46,XY[11]
ND
ND
48,XY,add(8)(p?11.2),der(14)
t(8;14)(q24;q32),+marx2[20]/48
,idem,del(12)(p1?3)[10]
1a
2b
3
4a
5b
6a
7a
8a
+
rear
(86%)
ND
ND
+
dup
(48%)
ND
ND
―
ND
MYC dup/rear
(% of cells
+
hd
(95%)
ND
ND
―
―
ND
―
CDKN2A
del
(% of cells)
ND
+
(95%)
ND
ND
+
(16%)
―
ND
―
IGH
rear
(% of cells)
ND
―
ND
ND
+
(32%)
―
ND
―
TP53
del
(% of cells)
ND
―
ND
ND
―
+
(37%)
ND
―
ATM
del
(% of cells
ND
―
ND
ND
ND
―
ND
―
cen12
tris
(% of cells)
ND
―
ND
ND
ND
―
ND
―
13q14
del
(% of cells)
ND
40~46,XX,-4[3],-6[5],-8[4],add(9)(p13)[5],-10[5], del(11)(q23)[5],-13[4],17[5],+4~7mar [cp6]/ 69~96,idem x2,-1[3],-2[3],-5[3],-11[4],+13[3], -18[3]
[cp5]/46,XX [10]
48,XY,add(8)(p?11.2),der(14)t(8;14)(q24;q32), +marx2[7]/48,idem,del(12)(p1?3)[3]
46~47,XY,add(14)(q32),+add(18)(q2?),+mar[cp7]/46,XY[4]
38~47,XY,add(1)(p3?4)[7],-9[12], der(9)del(9)(p2?)del(9)(q1?3)[12],add(9)(q?13)
[20], del(11) (q?21q?23)[14],-17[5],+18[3],-21[8], +22[3], +der(?;?)(?→?cen→?::?
→?cen→?::3q13→3qter) [24][cp26]
ND
44~48,XX,-1,add(6)(q1?5), del(7)(q32), +del(7)(q32),-9, del(12)(q13),add(17)
(p11),-21, der(?) t(?;1)(?;p13), der(?;?)(?→?cen→?::?→?cen→?::12q13→12q
ter),+3~4mar[cp14]
42~46,XX,der(1)(pter→q32::?q32→?q21::?q32→?q21:)[8], -2[3],add(3)(q21)
[8],-4[3],add(7) (q3?6)[4],-9[8], add(12)(p1?2)[7], +15 [6],-17[4],-18[4],-19[5],
21[5],+3~4mar[7] [cp8] /46,XX[4]
46,XY,del(2)(q23 or q31)[7]/ 47,sl,+3[2]/ 48,sdl1,+8[8]/46,XY[4]
Karyotype
RS
+
rear
(70%)
+
dup
(17%)
+
dup
(24%)
+
dup
(25%)
+
dup,amp
(99%)
+
rear
(14%)
+
rear
(7%)
ND
MYC dup/rear
(% of cells)
RS
+
hd
(80%)
ND
―
+
hd
(81%)
―
+
hd
(38%)
+
(43%)
CDKN2A
del
(% of cells)
+
( 37%)
+
(80%)
ND
+
(7%)
+
(11%)
―
―
―
IGH
rear
(% of cells)
+
( 54%)
RS
RS
―
ND
ND
+
(10%)
―
―
―
TP53
del
(% of cells)
+
( 54%)
RS
―
ND
ND
―
+
(95%)
―
―
ATM
del
(% of cells)
―
RS
―
ND
ND
ND
―
―
―
cen12
tris
(% of cells)
―
RS
―
ND
ND
ND
―
―
―
13q14
del
(% of cells)
―
amp – amplification; del – deletion; dup - duplication;hd – homozygous deletion; ND - not done; rear – rearrangement; ― - no abnormality; + - abnormality; “P” was defined as the progression stage that shows sometimes an increase in cell size and proliferative activity (but not CD71+++) as well as immunophenotyping changes (decrease CD62L and/or CD52 expression) in relation to the CLL/SLL.
Karyotype
No.
Table 3. Karyotype and FISH data of Richter syndrome patients and earlier stages of disease in some cases
CLL/SLL(a) or P CLL/ CLL/SLL(a) or CLL/SLL(a) or CLL/SLL(a) or CLL/SLL(a) or CLL/SLL(a) or CLL/SLL(a) or CLL/SLL(a) or
SLL (b)
P CLL/SLL (b) P CLL/SLL (b) P CLL/SLL (b) P CLL/SLL (b) P CLL/SLL (b) P CLL/SLL (b) P CLL/SLL (b) RS
CD71: (+++) - expression on 100% of cells; CYT a: [↑] - diffuse large B-cell lymphoma morphology; CYT b: CB - large cell size, cell polymorphism, abundant cytoplasm, irregular nuclear shape, multiple, small, excentric (centroblast-like) nucleoli, IB - large cell size, cell polymorphism, abundant cytoplasm, regular nuclear
shape, single, large, central (immunoblast-like) nucleolus, LB - medium cell size, usually not abundant cytoplasm, irregular nuclear shape, finely, granular (lymphoblast-like) nuclear chromatin; FSC/SSC: [↑] - increased cell size in comparison with smaller SLL/CLL cells; ND - not done; No. - case number; [+] - expression
on 100% of cells; [-] - no expression (<20% neoplastic cells); [+/-] - expression on >20%<100% of cells; [+↑] - positive expression, but higher in comparison with SLL/CLL cells or normal lymphocytes; [+↓] - positive expression, but weaker in comparison with SLL/CLL cells or normal lymphocytes; [+/-↓] - expression on
subpopulation of cells, but weaker in comparison with SLL/CLL; [-↓] - loss of antigen expression in comparison with positive expression on SLL/CLL cells; [+]bright or [+]dim - bright or dim expression of CD20; [+/-]dim - dim expression of CD20 on >20%<100% of cells; CD19>CD20: [+] the MFI of CD19 expression higher than
MFI of CD20 on neoplastic cells, [-] the MFI of CD19 expression weaker than MFI of CD20 on neoplastic cells.
↑
↑
2
1
FSC
No. CYT a CYT b /SSC
coexistence
of DLBCL
cells, and
CLL/ SLL
Table 2. Cytology, flow cytometry, morphology and immunophenotyping of Richter Syndrome
th
CHRONIC LYMPHOPROLIFERATIVE DISORDERS – A TWO YEAR
STUDY FROM A TERTIARY CANCER CENTRE IN SOUTH INDIA
EXTRANODAL NK/T-CELL LYMPHOMA, NASAL TYPE: A
CLINICOPATHOLOGIC AND GENOTYPIC STUDY OF 76 CASES
FROM TAIWAN
Rekha A. Nair, Priya Mary Jacob, A.V. Jayasudha, Renu Sukumaran,
K.R. Anila, Sreejith Nair, Sruthi Prem
Departments of Pathology and Medical Oncology, Regional Cancer Centre, Trivandrum, Kerala
State, India
Background: This study comes from the State of Kerala, South India. Around 600
cases of lymphomas are diagnosed and treated every year in Regional Cancer
Centre, Trivandrum and around 70 to 80 cases of chronic lymphoproliferative disorders (CLPD) are diagnosed by flow cytometry annually.
Aim of the study: To analyse and categorize the CLPDs diagnosed by flow cytometry
over a two year period.
Subjects and Methods: This is a 2 year retrospective study from January 2012
to December 2013. Diagnosis was made on peripheral smear/ bone marrow(PB/
BM) morphology and immunophenotyping by flow cytometry. A 4 color FACSCalibur
machine was used (stain-lyse-wash technique). A total of 125 cases of CLPDs were
retrieved. The PB and BMA smears were stained with giemsa and myeloperoxidase
stains for morphologic evaluation.
Results: Out of a total of 125 cases, 106 (85%) cases were B cell CLPDs and 19
(15%) were T cell CLPDs. Chronic lymphocytic leukemia was the commonest ( 64%)
followed by mantle cell lymphoma ( 13%).CD200 is a useful marker in differentiating these 2 entities. There were 6 cases of marginal zone lymphoma including
4 cases of splenic marinal zone lymphoma. 4 cases of hairy cell leukemia were
diagnosed which were all positive for CD25 and CD103. 5 cases of large B cell
lymphomas were diagnosed among which one case was coexpressing CD5 also ( all
confirmed by tissue biopy). 4 cases of leukemic phase of follicular lymphoma were
diagnosed of which 3 cases showed a loss of CD10 (CD10 was positive in the lymphnode biopsy)(ref.2). Among the TCLPDs, Adult T cell leukemia was the commonest
accounting for 74% followed by 1 case each of sezary cell leukemia,aggressive NK
cell leukemia, CD4+T large granular lymphocytic leukemia (T LGL), CD8+TLGL, anaplastic large cell lymphoma and enteropathy associated T cell lymphoma. Among
ATLLs,one case showed loss of CD5, CD7 and CD25. Another case showed expression of CD7 along with CD5 and CD25.(ref.3)
Summary and Discussion: In our study, commonest was CLL. CD 200 was a good
marker to differentiate CLL and mantle cell lymphoma in difficult cases. 5 cases of
blastoid and one case of pleomorphic mantle cell lymphomas were noted. In 3 out of
4 cases of follicular lymphoma, there was loss of CD10 by flow cytometry. Previously
we had shown that in seven out of 13 cases, there was absence of CD10 on flow
cytometric analysis of circulating cells.(ref 2).. Among the T cell CLPDS, a high number of ATLLs were diagnosed, all confirmed by positive serum HTLV1 status.(ref.3).A
few cases of unusual phenotypes among ATLLs were noted.
Conclusion: This study attempts to classify CLPDS into various subtypes based on
the current WHO classification system.(ref.1). Loss of CD 10 in circulating follicular
lymphoma cells and expression of CD5 in marginal zone lymphoma is noted. A high
number of ATLL cases is demonstrated with unusual immunotypes in a few cases.
References:
1. 2008 WHO Classification of Tumours of Haematopoietic and Lymphoid Tissue.
2.Priya Mary Jacob,Rekha A Nair et al.Downregulation of CD10 in leukaemic phase
of follicular lymphoma: a silent deception. J Hematopathol (2013) 6:65–70
3. Rekha A. Nair& Priya Mary Jacob et al Adult T cell leukaemia/lymphoma in
Kerala, South India: are we staring at the tip of the iceberg? J Hematopathol (2013)
6:135–144
Jie Yang Jhuang1, Sheng Tsung Chang2, Shih Feng Weng3,
Shien Tung Pan4, Pei Yi Chu5, Pin Pen Hsie6, Chih Hsin Wei7,
Shih Cheng Chou8, Chiew Loon Koo9, Chih Jung Chen10,
Jeng Dong Hsu11, Shih Sung Chuang12
1
Department of Anatomic Pathology, Far Eastern Hospital, New Taipei City, Taiwan
Department of Pathology, Chi-Mei Medical Center, Tainan, Taiwan, Department of Nursing, National
Tainan Institute of Nursing, Tainan
3
Department of Medical Research, Chi-Mei Medical Center, Tainan, Taiwan
4
Department of Pathology, Miaoli General Hospital, Miaoli, Taiwan
5
Department of Pathology, St. Martin De Porres Hospital, Chiayi, Taiwan and School of Medicine,
Fu-Jen Catholic University, New Taipei City, Taiwan
6
Departments Pathology and Laboratory Medicine, Kaohsiung Veterans General Hospital and Center
for General Education, Yuh-Ing Junior College of Health Care and Management, Kaohsiung, Taiwan
7
Department of Hemato-oncology, National Taiwan University Hospital, Hsin-Chu branch, Hsin-Chu,
Taiwan
8
Department of Pathology, Yuan’s General Hospital, Kaohsiung, Taiwan
9
Department of Pathology, Tungs’ Taichung MetroHarbor Hospital, Taichung, Taiwan
10
Department of Surgical Pathology, Changhua Christian Hospital, Changhua, Taiwan; School of
Medicine, Chung Shan Medical University, Taichuang, Taiwan; Department of Medical Technology,
Jen-Teh Junior College of Medicine, Nursing and Management, Miaoli, Taiwan
11
Department of Pathology, Chung Shan Medical University Hospital and School of Medicine, Chung
Shan Medical University, Taichung, Taiwan
12
Department of Pathology, Chi-Mei Medical Center, Tainan, Taipei Medical University and National
Taiwan University, Taipei, Taiwan
2
Extranodal nature killer (NK)/T-cell lymphoma (ENKTL), nasal type is a predominantly extranodal lymphoma associated with Epstein-Barr virus (EBV) and occurs most
often in the upper aerodigestive tract. The majority of cases appear to be genuine
NK-cell neoplasms, some cases may originate from cytotoxic T-cells, of either 
or  phenotype. There are very limited data on large cohorts of ENKTL in terms of
cellular lineages and their prognostic impact. We retrospectively investigated the
in-house and consultation cases at a single institute in southern Taiwan. A total of
76 cases were identified from January 1991 to December 2013. All patients were
Taiwanese with a median age of 54.5 years old and a M:F ratio of 2.0:1. The upper aerodigestive tract (designated as nasal group) was the most common site of
presentation (52 cases; 68%). The other presenting sites (n=24; non-nasal group)
included the skin, gastrointestinal tract, liver and bone marrow. Of the 68 cases with
complete staging, 49 cases (72%) had stage I or II disease while nineteen (28%)
had advanced (stage III/IV) disease. All but 1 (99%) and 5 cases (93%) expressed
CD3 and at least one cytotoxic marker, respectively. CD56 was expressed in 76%
(57/75) cases. All cases were positive for EBER. Using immunohistochemistry and
T-cell clonality analysis, these tumors were classified into NK-cell lineage (n=40;
53%), T-cell (n=14; 18%), and indeterminate (n=22; 29%). The only clinicopathological difference among these groups was CD5-negativity in the NK-cell group
(p=0.016). The overall survival time was shorter in the non-nasal group (Log-Rank
test; p =0.004), although there were no statistical difference in age, sex, histologic
or immunophenotypic features between nasal and non-nasal groups. We concluded
that nasal presentation accounted for nearly 70% of all ENKTL cases. Excluding
the cases with indeterminate lineage, 74% cases were of NK-lineage and 26%,
T-lineage. Non-nasal tumors were more aggressive than those with nasal presentation. A prospective national study is warranted for a better understanding of the
clinicopathological and genetic features of this uncommon tumor and the prognostic
factors.
[PP-LYMP-050]
[PP-LYMP-049]
Keywords: Epstein-Barr virus; EBER; TCR gene rearrangement; extranodal natural
killer/T-cell lymphoma, nasal type; NK/T-cell lymphoma; Taiwan
Keywords: CLPD, Flowcytometry, CLL
| 17-22 October 2014
| EAHP - 2014
|
103
th
[PP-LYMP-051]
THE INCIDENCE AND CLINICAL RELEVANCE OF BCL-2
NEGATIVITY IN FOLLICULAR LYMPHOMAS
Ayca Ersen1, Mehmet Ali Ozcan2, Aybuke Olgun3, Hulya Ellidokuz4,
Sermin Ozkal1
1
Dokuz Eylül University, Faculty of Medicine, Pathology Department, İzmir, Turkey
Dokuz Eylül University, Faculty of Medicine, Haematology Department, İzmir, Turkey
Dokuz Eylül University, Faculty of Medicine, Department of Internal Medicine, İzmir, Turkey
4
Dokuz Eylül University, Faculty of Medicine, Oncology Institute, İzmir, Turkey
2
3
Follicular lymphoma (FL) accounts for 20% of all lymphomas. BCL-2 positivity is an
important diagnostic clue. However about 10% to 15% of FL are BCL-2 negative,
most of which also lack the typical t(14;18). Although the extent of BCL-2 staining in
low grade FL cases is larger, no association between BCL-2 negativity and prognosis has been reported in FL so far. In this study we aimed to analyze the incidence
of BCL-2 negativity both with immunohistochemistry(IHC) by either conventional or
alternative antibody, also with flourescent in situ hybridization (FISH); and the impact
of BCL-2 negativity on clinical outcome.
We included 28 FL cases, diagnosed and treated in Dokuz Eylul University Hospital.
IHC staining was performed using both the conventional BCL-2 antibody from DAKO
and an alternative antibody E17 from Abcam. BCL-2 gene status was assessed
using a commercially available dual-color, Miksish 18q21 DNA split FISH probe (
Medimiks Biotechnology, Turkey).
Figure 1. Flow cytometry, pathological and immunophenotypical findings of
extranodal nasal type NK/T cell lymphoma.
The mean age of the patients was 62 years (range 35-84). Five of the cases
were extranodal. Eight cases also had bone marrow involvement (BMI). Ten
cases were at early stage. Two cases were followed without any treatment, 5
recieved mono-radiation therapy (RT), 10 recieved both chemotherapy (CT) and
RT, 11 cases recieved only CT. Twenty two patients recieved Rituximab based
chemo-immunotherapy.
The mean follow-up time was 60 months (range 9-149 months). Eight patients experienced relaps. Currently, only 1 patient has progressive disease, 2 patients are
still on CT regimen and the rest are alive and stable.
Relaps rate did not have any significant correlation with any clinical parameter. The
IHC results with the 2 antibodies were parallel. Ten cases were immunonegative.
These did not show any detectable BCL-2 break by FISH either.
Of the 18 BCL-2 positive cases, 8 experienced relaps whereas the number of cases
with relaps was only 1 among BCL-2 negative cases. Yet the difference was not
statistically significant.
Figure 2. The median time of T-cell lineage patients was 11 months. The 1-,
2-, and 5- year rates of the entire cohort were 48.5% (95% CI: 19.5-73.8%), 37.1%
(95% CI: 10.4-64.7%), and 18.5% (95% CI: 1.3%-52.3%) respectively. The median
time of NK lineage patients was 15 months. The 1-, 2-, and 5- year rates of the
entire cohort were 52.2% (95% CI: 34.8-67.1%), 50.0% (95% CI: 31.7-64.2%),
and 45.2% (95% CI: 28.0%-61.0%) respectively. The median time of indeterminate
lineage patients was 8 months. The 1-, 2-, and 5- year rates of the entire cohort
were 47.8% (95% CI: 24.8-67.7%), 42.5% (95% CI: 20.7-62.9%), and 26.6% (95%
CI: 9.7%-47.1%) respectively. Log-Rank P-value=0.762.
The mean relaps free survival (RFS) was 47 months. There was no statistically significant impact of age, gender, BMI, grade, stage on RFS. The BCL-2 negative cases
had a longer RFS. However the difference was not significant (p=0.078). The cases
with diffuse pattern experienced relaps earlier, yet again the difference was not
statistically significant (p=0.06)
Aberrant expression of the BCL-2 is an important diagnostic clue of FL. However,
in the reported results of Western Europe, about 10% to 15% of FL are BCL-2
negative and they also lack the typical t(14;18). The incidence of BCL-2 negativity in our series was 35% which is much higher than the literature. Asian
and Eastern Europe results are not well documented, and limited studies show
some inconsistent results of FL incidences and t(14;18) frequencies. This might
be due to an alternative pathogenetic pathway. Some studies also suggest that,
with conventional antibody, some BCL-2 negative cases are immunoreactant with
alternative antibodies like E17. We also performed the alternative antibody E17.
The results were parallel although the expression was much stronger and the
differentiation between non-neoplastic T-cells and neoplastic B-cells was much
more confidently done.
Our study shows that relaps was more seldomly encountered and the RFS was
longer in BCL-2 negative cases, however the results were not statistically significant
which maybe due to the limited number of cases involved in the study.
Keywords: Follicular lymphoma, BCL-2
Figure 3. The median time of nasal group patients was 25 months. The 1-, 2-,
and 5- year rates of the entire cohort were 61.6% (95% CI: 45.6-74.2%), 54.1%
(95% CI: 38.1-67.6%), and 42.6% (95% CI: 27.0%-57.3%) respectively. The
median time of non-nasal group patients was 4 months. The 1- and 2- rates of the
entire cohort were 27.5% (95% CI: 10.9-47.1%), and 27.5% (95% CI: 10.9-47.1%),
respectively. Log-Rank P-value=0.004.
104
| EAHP - 2014
| 17-22 October 2014
Joan Somja1, Laurence de Leval2, Ayse U. Akarca1,
Christiane Copie Bergman3, Teresa Marafioti1, Philippe Gaulard3
1
2
3
Figure 1. BCL-2 negative FL case. a. FL case (H&E, 40x) b. BCL-2 negative on
E17 antibody (40x) c. CD10 positivity of the tumor cells (100x) d. BCL-2 break is not
detected on FISH
[PP-LYMP-052]
COMPOSITE CLASSICAL HODGKIN LYMPHOMA AND
FOLLICULAR LYMPHOMA MAY DISPLAY BOTH BCL2 AND
BCL6 GENE REARRANGEMENT IN NEOPLASTIC CELLS BY
FLUORESCENCE IN SITU HYBRIDIZATION
Vanessa Szablewski1, Maryse Baia2, Marie Hélène Delfau Larue3,
Corinne Haioun4, Taoufic Elgnaoui4, Philippe Gaulard5,
Christiane Copie Bergman5
1
Département de Biopathologie Cellulaire et Tissulaire des Tumeurs, CHU Montpellier, Hôpital Gui
De Chauliac, Montpellier, France
2
3
4
5
Classical Hodgkin Lymphoma (cHL) and Follicular lymphoma (FL) are distinct disease entities, however reports on composite cHL and FL lymphomas have been
repeatedly described. These cases represent models to study the multistep transformation process of cHL and FL and suggest that both diseases may in some instances originate from a common precursor cell. In the present study, we report
5 patients (4M/1F) with a mean age of 72 years [55-81], who presented with cHL
and FL occurring in the same anatomic site (n=3) or synchronously in separate
sites (n=2). cHL cases showed typical morphological features of nodular sclerosing
cHL and Hodgkin/Reed Sternberg (HRS) cells displayed an CD20-, PAX5+, CD3-,
CD30+, CD15+/-, EBV- immunophenotype. FL had a follicular growth pattern, were
composed of centrocytes admixed with centroblasts (grade 1-2) and tumor cells
were CD20+, CD5-, CD10+, BCL2+. Fluorescence in situ hybridization (FISH) using
BCL2, BCL6 breakapart probes and IGH-BCL2 fusion probe was performed in all
cases as well as in 19 cases of cHL without any associated FL as a control group.
The t(14;18)(IGH-BCL2) translocation was observed by FISH in four of the five cases
in both FL tumor cells and HRS cells. In addition, two cases showed both BCL2 and
BCL6 gene rearrangements in HRS and FL neoplastic cells. In the control group, 2
cHL cases displayed BCL6 gene rearrangement in HRS cells whereas BCL2 gene
rearrangement was not observed in any case. The demonstration for the first time
of BCL2 and/or BCL6 alterations in situ in both HRS and FL cell components by FISH
analysis further supports a possible common origin of both components. It can be
speculated that a subset of cHL may evolve from a pre-existing FL with the t(14;18)
and or BCL6 rearrangement. Whether the t(14;18) may represent merely one process among several that predispose to cHL development is questionable.
Department of Histopathology, University College London Hospital, London, United Kingdom
University Institute of Pathology, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland
Department of Pathology, Hôpital Henri Mondor, Créteil, France
CD103 (7 integrin) is an adhesion molecule playing a major role in T-cell migration and epidermotropism. CD103 expression determined by frozen section
immunohistochemistry and/or flow cytometry has been reported in enteropathyassociated T-cell lymphomas (EATL) and cutaneous T-cell lymphomas but in other
T-cell lymphoma (TCL) entities is still poorly documented. The aim of this study
was to analyze the expression of CD103 by immunohistochemistry using the rabbit
monoclonal anti-CD103 antibody (clone EPR4166 Epitomics, Burlingame, CA) on
(routinely-fixed) paraffin-embedded tissue sections of a large series of TCL (n=109)
and assess its diagnostic utility. Immunostainig was evaluated by three expert haematopathologists using the following score criteria: 0: all cells negative; 1: <20%
positive tumour cells (TC); 2: 20%-50% positive TC and 3: >50% positive TC. CD103
was found in >20% TC in 11/24 EATL and 2/2 mycosis fungoides (MF) in association with the epitheliotropic or epidermotropic component in several cases whereas
in 10 EATL cases CD103 staining was observed in a small proportion of cells with
uncertain neoplastic nature (Table 1). In addition, in 6 out of the 11 CD103-positive
EATL cases as well as in a few cases of PTCL, not otherwise specified, in adult T-cell
leukaemia/lymphoma and in extranodal NK/T cell lymphoma, an heterogeneous expression of CD103 was observed (>20% but <100% of TC) (Table 1). In a total of 15
out of 109 TCL, highlighted with an asterisk in Table 1, CD103-positive cells showed
equivocal morphology between reactive and neoplastic T-cells. To better characterize those cells, double immunostaining is in progress. Large granular lymphocytic
leukaemia, hepatosplenic T-cell lymphoma and T-acute lymphoblastic leukaemia/
lymphoma were CD103-negative. This is the first description of CD103 in a large
series of TCL. Our results confirm the diagnostic relevance of CD103 in EATL and MF
and highlight its expression confined to a small proportion of cases in several entities. The findings might reflect distinct migration capacities and biological behaviors
of these lymphomas. The functional meaning of the heterogenous expression of
CD103 prompts future molecular studies.
SELECTIVE EXPRESSION OF CD103 IN T-CELL LYMPHOMA
ENTITIES
[PP-LYMP-053]
Keywords: CD103, T-cell lymphoma, immunohistochemistry
Keywords: Composite, follicular lymphoma, classical Hodgkin lymphoma
| 17-22 October 2014
| EAHP - 2014
|
105
NUCLEAR OPTICAL DENSITY IS HIGHER IN BURKITT’S
LYMPHOMA THAN IN DIFFUSE LARGE B-CELL LYMPHOMA
Zaklina Zarko Mijovic, Dragan Slavoljub Mihailovic
Centre of Pathology, University of Nis, Serbia
Introduction: Immunohistochemistry and molecular biology are mandatory for diagnosing diffuse large B-cell lymphoma (DLBCL). On other hand, Burkitt’s lymphoma
has a highly aggressive course but is curable by polychemotherapy.
Aim: To estimate karyometric variables in diffuse large B-cell lymphoma (DLBCL)
and Burkitt’s lymphoma.
Material and Methods: At Centre of Pathology, Clinical Centre of Nis, 7 cases of
DLBCLs and 3 cases of Burkitt’s lymphomas were diagnosed using appropriate
histological and immunohistochemical criteria. Using ImageJ software, on digital
images obtained at x40 objective, seven nuclear variables were estimated: nuclear
area, mean optical density (OD), modal optical density, perimeter, Feret diameter,
integrated optical density (IOD) and roundness.
Results: Modal nuclear optical density was significantly higher in Burkitt’s lymphoma than in DLBCL. In contrast, nuclear area, perimeter and Feret diameter were
significantly higher in DLBCL than in Burkitt’s lymphoma. Differences in nuclear
shape were not statistically significant.
Conclusions: In individual cases, the distinction of Burkitt’s lymphoma from DLBCL
can be more precise using karyometry.
Keywords: Lymphoma, karyometry, density
th
[PP-LYMP-054]
Keywords: diffuse large B-cell lymphoma; fluorescent in situ hybridization; intestine; perforation; Taiwan
[PP-LYMP-056]
MYC ONCOGENE IS DIFFERENTIALLY EXPRESSED AND
CORRELATES WITH P53 EXPRESSION LEVELS IN B-CELL NON
HODGKIN LYMPHOMAS
Anna Kwiecinska1, Mehran Ghaderi1, Ioanna Xagoraris1,
Leonie Saft1, Elias Drakos3, Efstratios Patsouris2,
George Z. Rassidakis1
1
[PP-LYMP-055]
Shih Sung Chuang1, Sheng Tsung Chang1, Sheau Fang Yang2,
Wan Ting Huang3, Pin Pen Hsieh4, Jeng Dong Hsu5,
Shih Feng Weng6, Mei Hua Tsou7
1
Pathology, Chi-Mei Medical Centre, Tainan, Taiwan
Pathology, Kaohsiung Medical University Hospital and Kaohsiung Medical University, Kaohsiung,
Taiwan
3
Pathology, Chang Gung Memorial Hospital – Kaohsiung Medical Centre and Chang Gung
University College of Medicine, Kaohsiung, Taiwan
4
Pathology, Veterans General Hospital-Kaohsiung, Kaohsiung, Taiwan 6Division of Hematooncology, Department of Internal Medicine, Chi-Mei Medical Centre, Tainan, Taiwan
5
Pathology, Chung Shan Medical University Hospital and School of Medicine, Chung Shan Medical
University, Taichung, Taiwan
6
Medical Research, Chi-Mei Medical Centre, Tainan, Taiwan
7
Pathology & Laboratory Medicine, Koo Foundation Sun Yat-Sen Cancer Center, Taipei, Taiwan
2
The gastrointestinal tract is the most common site of primary extranodal nonHodgkin lymphoma; with mucosa-associated lymphoid tissue (MALT) lymphoma
and diffuse large B-cell lymphoma (DLBCL) as the most frequent histological
types. Using cDNA microarray method, DLBLs could be divided into prognostically
important subgroups and immunohistochemistry for differentiation antigens may
serve as surrogate markers. There are only a few reports on the genetic and/or
fluorescent in situ hybridization (FISH) data and their prognostic impact on primary intestinal DLBCL (PI-DLBCL). To investigate the prognostic impact of depth of
tumor invasion including perforation, differentiation antigens and phenotype, and
chromosomal translocations of PI-DLBCLs, we retrospectively collected 59 cases
from Taiwan from January 1991 to December 2013. We reviewed histopathology,
performed immunohistochemistry and FISH and reviewed the medical records.
There were 31 males and 28 female with a median age of 66 years (range, 2388). Eleven (19%) tumors were perforated at presentation. Eight (14%) patients
had multicentric tumors. The ileum (24 cases; 41%) and ileocecum (6 cases;
10%) were most frequently involved sites. There were 22 (37%) patients at stage
IE, 31 (53%) at stage IIE, 1 (2%) at stage III and 5 (8%) at stage IVE, respectively.
All patients received operation including right hemicolectomy (30 patients; 51%),
segmental resection (26; 44%), Whipple operation (2; 3%), and appendectomy (1;
2%). Twenty one (36%) patients did not receive chemo- or radiotherapy including
6 patients who presented with perforation and died within 0.2 to 7 months after
| EAHP - 2014
| 17-22 October 2014
Department of Pathology and Cytology, Karolinska University Hospital, Stockholm, Sweden
First Department of Pathology, National and Kapodistrian University of Athens Medical School,
Athens, Greece
3
Department of Pathology, University of Crete Medical School, Heraklion Crete, Greece
2
PERFORATION BUT NOT IMMUNOPHENOTYPE OR
CHROMOSOMAL TRANSLOCATION PREDICTS A POOR
PROGNOSIS IN PRIMARY INTESTINAL DIFFUSE LARGE B-CELL
LYMPHOMA
106
operation. Thirty-eight (64%) patients received chemotherapy and seven (12%)
with additional radiotherapy. The 1-, 2-, and 5-year overall survival rates of the
entire cohort were 68.4%, 56.5% and 50.0%, respectively. Eighteen (31%) tumors
were of germinal center B-cell (GCB) phenotype and the remaining 41 (69%), nonGCB type according to the Hans’ algorithm. Myc protein expression was noted in
31 (53%) tumors and was not statistically associated with survival (p =0.166).
Rearrangement at the IGH, BCL2, BCL6, and MYC foci were identified in 22%
(13/59), 3% (2/59), 17% (10/59), and 7% (4/58) cases, respectively. Eight (14%)
of 56 cases showed gain/amplification at the MYC locus and was not statistically
associated with survival (p =0.325). In this series of PI-DLBCL, intestinal perforation at presentation (p = 0.009), high ECOG PS (3 and 4) (p=0.018) and without
adjuvant chemotherapy (p<0.001) were poor prognostic factors, but not GCB or
non-GCB phenotypes or lymphoma-associated chromosomal translocations. In
conclusion, we characterized the clinicopathological and molecular features of
PI-DLBCL in Taiwan and found a relatively higher rate of perforation and lower
frequency of GCB phenotype as compared to other geographic areas.
Background: MYC is an oncogene and potent transcription factor involved in cell
cycle deregulation. MYC is overexpressed due to t(8;14)(q24;q32), which is detected in Burkitt lymphoma (BL) and in other aggressive non-Hodgkin lymphomas
(NHL) including a subset of diffuse large B-cell lymphomas (DLBCL). In addition,
MYC oncoprotein can be upregulated in hematologic malignancies in the absence
of chromosomal aberrations involving the 8q24 locus. Detection of MYC protein
using routinely processed paraffin-embedded tissues has recently been validated
because of the availability of reliable anti-MYC antibodies.
Purpose: This study aimed to systematically investigate the expression levels of
MYC in a series of indolent and aggressive B-cell NHL by immunohistochemistry
using a validated antibody and correlate the findings with p53 expression and tumor
cell proliferation.
Methods: A series of 240 B-cell NHLs including 74 indolent and 166 aggressive
lymphomas were included in the study. In addition, 72 classical Hodgkin lymphomas, 8 cases of t(8;14)-positive BL as well as 5 reactive lymph nodes were also
analyzed for comparison. The DLBCLs with very high cell proliferation (>90%)
included in the study were previously tested negative for 8q24 chromosomal aberrations by FISH. The antibody used for MYC (MAb Y69) has recently been validated for immunohistochemical studies (G. Cattoretti, J Pathol 2013; 229:430).
p53 expression was assessed using the DO-7 antibody that detects both the wildtype and mutated p53 gene products. Any nuclear staining in tumor cells was
considered positive irrespective of intensity for both MYC and p53. In selected
cases, p53 mutation analysis was performed for the exons 4-8 of the gene using
PCR and direct sequencing techniques. Proliferation index was assessed using
the Ki67 (MIB-1) marker.
Results: In reactive lymph nodes, MYC was detected in the nucleus of a small
subset (approximately 2-10%) of germinal center (GC) cells mostly localized at the
periphery of the GCs, but it was also occasionally found in small lymphocytes in interfollicular areas. The mean percentage of MYC-positive neoplastic cells varied significantly among the B-cell NHL types being 5.2% in indolent lymphomas, 41.4% in
aggressive lymphomas, and 81.5% in cHL (p<0.001, Kruskal Wallis test). Similarly,
the mean percentage of p53-positive tumor cells was 9.9% in indolent lymphomas,
40.6% in aggressive lymphomas and 79.3% in cHL (p<0.001, Kruskal Wallis test). In
the entire B-cell NHL study group, MYC levels were significantly associated with p53
levels (Spearman R 0.66, p<0.001) and Ki67 cell proliferation index (Spearman R
0.39, p=0.004). However, the association between the percentage of MYC-positive
tumor cells and Ki67 index was weaker and did not reach statistical significance
within the aggressive B-cell NHL subgroup (Spearman R 0.26, p=0.07). As a positive
THE STORY OF A HAIRY CELL: HOW NECESSARY IS BRAF
ACTUALLY?
Tihomir Dikov, Yavor Topalov, Gueorgui Balatzenko,
Margarita Guenova
National Specialised Hospital for Active Treatment of Haematological Diseases, Sofia, Bulgaria
Keywords: B-cell non Hodgkin lymphomas, MYC, p53
[PP-LYMP-057]
FLOW CYTOMETRIC ANALYSIS OF SOX11; A NEW DIAGNOSTIC
METHOD FOR DISTINGUISHING B-CLL FROM MCL
Agata M. Wasik, Valdemar Priebe, Martin Lord,
Åsa Jeppson Ahlberg, Birger Christensson, Birgitta Sander
Laboratory Medicine, Pathology, Karolinska Institutet and Karolinska University Hospital Huddinge,
Stockholm Sweden
Introduction: Flow cytometric analysis is a complimentary method in diagnosis
and subtyping of non-Hodgkin lymphomas. Mantle cell lymphoma (MCL) and B-cell
chronic lymphocytic leukemia/small cell lymphocytic lymphoma (B-CLL/SLL) are
mature B-cell lymphomas that may present with lymphocytosis. The differential
diagnosis between these entities is essential since MCL usually has a more aggressive clinical course. By flow cytometry both MCL and B-CLL are CD19, CD5 and
CD20 positive. In contrast to B-CLL/SLL most MCL are negative for CD23, however
a subset of MCL is CD23 positive. Also, other ambiguities in the phenotypic distinction of MCL from B-CLL/SLL and other B-cell lymphomas occur. The transcription
factor SOX11 is highly upregulated in most MCL, including t(11;14)(q13;q32) negative cases.
Here, we investigated if SOX11 expression could be used in flow cytometry to
discriminate MCL from B-CLL using the new monoclonal anti-SOX11 antibody
(MRQ-58).
Methods: Basal protein expression of SOX11 was verified by western blot and flow
cytometry using the commercially available mouse monoclonal anti-SOX11 antibody MRQ-58 in six MCL cell lines: Granta519, Rec1, JeKo1, Mino, Z138 and JVM2.
Antibody specificity was verified by transient knockdown of SOX11 in Granta519
and Z138 cell lines and immunoreactivity in the nucleus was confirmed by confocal
microscopy. SOX11 expression was investigated in 11 tumor samples of primary cyclin D1-positive MCL from 10 patients and 10 tumor samples of B-CLL by multicolor
flow cytometry. In all samples SOX11 mRNA levels were also determined by qPCR.
Results: Immunoblotting for SOX11 showed that the antibody used detected a single
band at the predicted size for SOX11 protein in SOX11 positive MCL cell lines, but
did not bind to any protein in the SOX11 negative MCL cell line, JVM2. Similar results
were obtained using flow cytometry: five SOX11 positive cell lines showed higher
SOX11 fluorescence intensity compared to the isotype control, whereas for JVM2
the SOX11 fluorescence intensity overlapped with the isotype control. Multicolor
flow cytometry showed that all primary MCL tested were SOX11 positive with median fluorescence intensity (MFI), after subtraction to isotype control, in the range
12.63 to 87.26 SOX11 was not detected in any of the B-CLL cases by flow cytometry
(MFI range -2.25 to 1.24). We correlated MFI to the Ct values obtained from the
qPCR results (mean of triplicates). Since we did not relate SOX11 protein levels to
any other protein but to the isotype control, we used equal amounts of cDNA within
the samples set (the higher Ct the lower mRNA expression). We found that those two
parameters correlated, Spearman correlation coefficient -0.69 (p=0.002).
Conclusion: We showed that SOX11 protein consistently could be detected and
quantified by flow cytometry. Implementing detection of SOX11 in the routine diagnostic flow cytometry would be beneficial for accurate and reliable diagnosis
of MCL, especially for phenotypically aberrant cases. SOX11 expression also helps
distinguish MCL from other malignancies with similar phenotype, like B-CLL. In addition, analyzing SOX11 by flow cytometry will generate quantitative answers. The
possibility of using SOX11 in a multi-color panel with other markers is an additional
advantage.
In the year 2009, a man aged 56 at that time, suffered severe traumatic event
and was immediately admitted in a hospital where initial workup showed excessive
splenomegaly combined with leukocytosis – 26 G/L. A hemathology consultation
ensued and ended up with 20 % morphologically recognizable hairy cells in peripheral blood.
A bone marrow biopsy showed hypercellular bone marrow with discernible diffuse interstitial infiltrates of medium sized lymphoid cells with pale cytoplasmic
rim and clearly defined cytoplasmic borders. This was judged to be consistent
with hairy cell leukemia (HCL). At that time no immunohystochemistry (IHC) was
preformed.
Symptom relieving splenectomy was performed the following month to reveal
that normal structure was obliterated by proliferation of small to moderate atypical lymphocytes, overfilling the red pulp cords with dispersed red blood cell
lakes.
Shortly after, the patient presented with leukocyte count 72 G/L and immunophenotyping by flow cytometry (FCM) of peripheral blood showed 53.5 G/L lymphocytes
with CD19+CD20bright+CD22bright+FMC7+CD103+CD11c+ phenotype, some of
them expressing low levels of CD25. The combination with prolymphocytic morphology proved evidence of HCL – variant (vHCL), followed by Fludara + Endoxan
chemotherapy regimen. At the end of treatment cycle, however, enlarging cervical
lymph nodes were detected. This, naturally, made the clinical team suspicious of the
correctness of the diagnosis and a biopsy was initiated since HCL is believed to not
be frequently associated with peripheral lymph node involvement.
Conclusions: Our findings show that MYC oncogene is overexpressed in aggressive
B-cell NHLs and cHL despite the apparent absence of MYC chromosomal aberrations. Upregulation of MYC is strongly associated with p53 levels, suggesting a
direct or indirect biologic link between the two transcription factors in lymphomas.
[PP-LYMP-058]
control group, BL showed high proportion of MYC-positive tumor cells (95-100%).
The lowest mean percentages of MYC and p53 positive tumor cells, 2.8% and 1.8%,
respectively, were observed in chronic lymphocytic leukemia / small lymphocytic
lymphoma.
Lymph node biopsy actually consisted of tiny pieces of tumor tissue made up of
small to medium sized atypical lymphocytes with barely visible nucleoli infiltrating
fragments of a major salivary gland. By IHC they turned out to be positive for CD20
and TRAP while negative for CD5 and CD10, Ki67 was low (<10%). As if this wasn’t
enough, the clinical team asked for immunophenotyping, which revealed abnormal
B-cell population in peripheral blood (2.9 G/L) positively expressing CD19, CD20,
CD22, CD103, CD123, CD200, FMC7 and showed no positivity for CD11c and CD25.
Thus morphological and phenotypic data confirmed the vHCL hypothesis and the
therapeutic regimen was switched to cladribin. The routine checkup showed 0.001
G/L cells with vHCL phenotype (CD103+CD25-CD11c+), more or less consistent
with a partial remission, which was short-lived since six months later the patient
presented with anemia (Hb 83 g/l), thrombocytopenia (Plt 25 G/L) and leukocytosis
(WBC 60 G/L). FCM identified 83.8% of lymphocytes to have CD19+CD20bright+C
D22bright+CD103+CD11c+CD81+ phenotype and surprisingly distinct CD25 expression appeared. Of note, BRAF testing was not widely known and hardly available
at that time.
At present, based on our recent experience, we retrieved archival trephine, spleen
and lymph node paraffin-embedded material and DNA was tested on all of them for
BRAF V600E mutation.
The case presented seems to be interesting for: 1) enlarged peripheral lymph nodes
with total effacement of structure, not conforming to previously described leukemic
type of infiltration, surrounding lymphoid follicles 2) triple positive confirmation of
HCL variant (CD20bright+CD103+ CD103+CD11c±CD25-) until CD25 unexpectedly
emerged, consistent with classical HCL (phenotypic shift vs. clonal evolution?). BRAF
testing seems highly promising to help in the situation.
Keywords: haitry cell BRAF
Keywords: SOX11, flow cytometry
| 17-22 October 2014
| EAHP - 2014
|
107
th
[PP-LYMP-059]
STATHMIN 1, COULD IT BE A MARKER FOR PROBLEM CASES
OF FOLLICULAR LYMPHOMA?
1
1
1
with MYC expression in the initial bone marrow. More sensitive tests for detection
of minor population of highly abnormal cells will provide insights into genetic basis
and early prediction for Richter transformation.
Keywords: chronic lymphocytic leukemia, Richter transformation
2
Seda Akturk , Gulsah Kaygusuz , Hale Kivrak , Kenan Kose ,
Yasemin Sahin1, Isinsu Kuzu1
1
Ankara University School of Medicine, Department of Pathology
Ankara University School of Medicine, Department of Biostatistics
2
Background: Follicular lymphoma is a challenging indolent lymphoma of follicle
centre B lymphocytes with heterogeneous morphology and immunophenotype.
About 60-90% of cases have t(14,18)(q32;q21) creating BCL2 rearrangement
whilst, 10-15% of cases carry 3q27 abnormality causing BCL6 rearrangement.
Besides histopathology, immunohistochemical positivity of BCL2, CD10 and BCL6
are helpful diagnostic markers. The expression rate of the characteristic markers
may vary with the histological grade, and this could complicate the accurate diagnosis. Stathmin 1 is a microtubule destabilizer and plays a critical role in the
regulation of mitosis. Recently, found to be overexpressed in many types of human
cancers. The aim of this study was to assess stathmin 1 status, its correlation with
clinicopathological parameters and with 1p36 gene rearrangement, and its role as a
diagnostic marker in Follicular lymphoma.
Methods: This study included 81 follicular lymphoma cases which Stathmin 1 expression was evaluated by immunohistochemistry on macroarray slides.
Figure 1. Bone marrow at initial presentation.
Results: Stathmin 1 was positive in 88,9% of cases. There was a positive correlation
between the histologic grade and Stathmin 1 expresion (p=0.001, Chi-Square test).
All BCL2- cases (n=13) were positive for Stathmin 1 whereas, 8/9 of CD10- cases
were positively stained.
Conclusion: Stathmin 1 was helpful in detecting BCL2- and CD10- follicular lymphomas, and it could be used as an helpful dianostic marker for those problematic
cases.
Keywords: Stathmin 1, Follicular Lymphoma
Figure 2. Bone marrow at transformation.
[PP-LYMP-060]
RICHTER TRANSFORMATION OF CHRONIC LYMPHOCYTIC
LEUKEMIA/SMALL LYMPHOCYTIC LYMPHOMA
Qing Ching Chen, Muhammad Adnan Malik, Zeba Singh
Department of Pathology, University of Maryland School of Medicine, Baltimore, USA
A 74-year-old man without significant past medical history presented with fatigue,
and was found to have anemia and mild lymphadenopathy. WBC count was normal
and there was no lymphocytosis. A bone marrow biopsy showed a hypercellular
marrow with extensive interstitial and nodular lymphoid infiltrate. Most of the lymphocytes were small to medium sized with clumped chromatin. However, a sizable
population (~10%) of large lymphocytes was present, which were positive for MYC
(Image 1). Flow cytometry identified a population of monoclonal B cells with a phenotype consistent with CLL. Cytogenetics showed an abnormal karyotype (Image 1).
FISH analysis detected 13q14 deletion. IgVH Mutation analysis showed unmutated
status. IgVH family usage was IgHV4-39 with a stereotype HCDR3. CGH analysis
identified a single copy loss of chromosome Y and a single copy gain in chromosome 12q. The patient was diagnosed with CLL/SLL and treated with Bendamustin
and Rituximab.
Figure 3. Lymph node at transformation.
Six weeks after chemotherapy, the patient’s condition worsened and was found to
have severe anemia, increased LDH and diffuse lymphadenopathy with the largest
one in the abdomen measuring 9.3 x 7 x 15 cm. A second bone marrow biopsy
showed marrow infiltrate by large lymphoid cells (Image 2). An inguinal lymph node
biopsy showed diffuse infiltrate by large B cells. Most of cells were positive for MYC
(Image 3). Conventional cytogenetics showed an abnormal karyotype, which is different from that seen in initial bone marrow (Image 3). FISH analysis showed 13q14
deletion and MYC rearrangement. A diagnosis of Richter transformation of CLL/SLL
was made. The patient was treated with reduced dose R-CHOP, and had a partial
response with reduced lymphadenopathy.
This case of CLL/SLL demonstrated a very rapid progression to Richter syndrome
despite of immunochemotherapy. It is currently unclear what factors predict rapid
large cell transformation. Several unusual features observed in this case may be associated with such transformation. These include lack of lymphocytosis, the usage
of unmutated VH4-39 with stereotype CDR3, and increased number of large cells
108
| EAHP - 2014
| 17-22 October 2014
MARGINAL ZONE LYMPHOMA WITH A SUBSET OF CD30 AND
CD15 POSITIVE REED-STERNBERG – LIKE LYMPHOID CELLS IN
A PATIENT WITH RHEUMATOID ARTHRITIS
Chad Ellermeier, Sonja Chen, Zakaria Grada, Metin Timur Mujdaba,
Diana Olguta Treaba
Department of Pathology and Laboratory Medicine, Lifespan Academic Medical University, The Alpert
School of Medicine at Brown University
Introduction Hodgkin-like transformation in a marginal zone lymphoma is an extremely rare event, with only a single case reported in the English medical literature
in the past 30 years (Fung EK et al, 2001). Case A 64-year old male developed
bulky axillary lymphadenopathy while receiving chronic immunosuppressive therapy (etanercept and methotrexate) for rheumatoid arthritis. CT imaging detected
bulky axillary and mild mediastinal lymphadenopathy. The patient also reported
fatigue, intermittent nocturnal fevers and night sweats. His CBC was notable for
a mild macrocytic anemia (MCV 99.8 fL, Hgb 12.3 g/dL), thrombocytopenia (127 x
109/L), and a normal white blood cells count. He underwent an excisional biopsy
of a right 6.9 x 2.6 x 2.0 axillary lymph node. Results The microscopic examination
of the right axillary lymph node was remarkable for diffuse effacement of the normal architecture by a polymorphic population of small lymphoid cells admixed with
histiocytes, eosinophils, scattered plasma cells and also scattered as single cells
or forming small loose clusters, large transformed lymphoid cells. These large lymphoid cells were mononucleated-immunoblast like or had multilobulated nuclei with
inclusion like nucleoli, reminiscent of Reed-Sternberg cells and their variants. By
immunohistochemistry, a larger subset of the lymphoid cells present (including the
large transformed lymphoid cells) were positive for B-cell markers: CD20, PAX5 and
CD79a, in a large subset were positive for CD45, p53, bcl-2 (variable) and the large
transformed lymphoid cells were also positive for CD30, MUM-1, bcl-6, and were
in a small subset CD15 positive. Admixed was a smaller subset of CD3+, bcl2+,
CD5+ T-cells. The Reed-Sternberg like cells and their variants were negative for TdT,
CD10, bcl-1, EMA, CD56, CD43, CD8, CD4, granzyme B and ALK1. A high proliferation index was highlighted by the immunoreactivity to the MIB-1 antibody, reaching 60-70% in areas. Molecular studies detected the presence of immunoglobulin
heavy chain rearrangements. Conclusions A diagnosis of marginal zone lymphoma
with a subset of CD30 and CD15 positive Reed-Sternberg – like lymphoid cells was
rendered, and the diagnosis was confirmed by expert consultation. While concomitant Hodgkin lymphoma and marginal zone lymphoma has been previously reported,
Hodgkin-like transformation in nodal marginal zone lymphoma is extremely rare and
in our patient it is also notable the chronic therapy with methotrexate that may have
contributed to this unusual morphology.
Keywords: marginal zone lymphoma, CD15, CD30
Figure 2. CD30.
[PP-LYMP-061]
Figure 3. Hematoxylin and eosin.
[PP-LYMP-062]
LOW-GRADE B-CELL LYMPHOMA WITH CEREBROSPINAL
INVOLVEMENT
Rashna Clubwala1, Heinrich Elinzano2, Peter Rintels3,
Diana Olguta Treaba1
1
Department of Pathology and Laboratory Medicine, Lifespan Academic Medical Center, The Alpert
School of Medicine at Brown University, Rhode Island
2
The Neurology Foundation, Lifespan Academic Medical Center, The Alpert School of Medicine at
Brown University, Rhode Island
3
Division of Hematology Oncology, Rhode Island Hospital, Rhode Island
Cerebrospinal fluid (CSF) involvement by low-grade B-cell lymphomas has a low
reported incidence (3%, Spectre G. et al, 2005) in the medical literature. We are
reporting two cases of low-grade B-cell lymphoma, one with involvement of the CSF
6 years after the original lymphoma diagnosis, and the second case with neurologic
symptoms and CSF involvement as disease presentation.
Figure 1. CD15.
The first patient is a 56-year old man, diagnosed 6 years ago on a right axillary lymph
node with a nodal marginal zone B-cell lymphoma. The neoplastic B-lymphoid population was surface immunoglobulin kappa light chain restricted and co-expressed
CD20, CD23, CD25, FMC7 being negative for CD5 and CD10. His bone marrow biopsy was characterized by extensive lymphomatous involvement (>95%) while his
peripheral blood was only remarkable for lymphopenia (absolute lymphocyte count
0.54 x 109/L). Despite repeated courses of chemotherapy, in the next 6 years the
patient had persistent and extensive bone marrow involvement by his lymphoma
and his bone marrow biopsies were also remarkable for increased reticulin deposition. The karyotype analysis showed in various biopsies a complex chromosomal
complement: 54,XY,+X,+del(3)(p21p25),+4,i(8)(q10),+9,+15,+16,+?18+,+21 associated with the clonal lymphoid population. During the years the patient became
pancytopenic and transfusion dependent. Due to a presentation with near syncope,
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The second patient is a 63-year old man with a history of hypertension, diabetes
mellitus and a recent history of altered mental status, ataxia and falls complicated
by a right shoulder fracture. A CSF fluid was remarkable for lymphocytosis with a
predominant population of small to medium size lymphoid cells, in a small subset
with a plasmacytoid appearance and by flow cytometry immunophenotypic analysis
a kappa light chain restricted B-lymphoid population in a small subset CD5+ was
detected. The patient’s CBC indices were within the reference ranges, however, the
flow cytometry analysis of patient’s peripheral blood was also remarkable for a kappa light chain restricted B-lymphoid population (absolute number 0.4 x 109/L), with
moderate CD20 and CD22 positivity, with FMC7 +, and with partial CD23 positivity,
but negative for CD5 and CD10. MRI analysis was remarkable for cerebellar, spinal
cord and cauda equina leptomeningeal enhancing lesions and for mediastinal and
hilar lymphadenopathy. As in the first case, CSF involvement by a low-grade B-cell
lymphoma, immunophenotypically favored to represent a marginal zone lymphoma
was considered. Both patients underwent intrathecal chemotherapy with cytarabine
and showed amelioration of their neurologic symptoms.
The cases presented raise attention to neurological symptoms as late or first manifestation of indolent B-cell lymphomas, and strongly suggest the use of ancillary
studies in the appropriate clinical setting to further characterize a CSF lymphocytosis.
Keywords: CSF, lymphoma, MRI
th
a CSF fluid was examined and was remarkable for lymphocytosis with atypical lymphoid cells of predominant small to medium size, and had also a few large sized
lymphoid cells. The flow cytometry analysis identified a kappa light chain restricted
B-lymphoid population, and by PCR the clonal band identified in the CSF appeared
similar in size to the one identified in the bone marrow biopsies. The MRI showed
leptomeningeal involvement of the right frontal, and left parieto-occipital regions
and diffuse enhancement of cranial nerves 3, 5, 7, 8 and 9.
[PP-LYMP-063]
HISTOLOGICAL AND IMMUNOHISTOCHEMICAL STUDY OF
MALIGNANT LYMPHOMAS IN MACEDONIA - STUDY OF 222
CASES
Gordana Petrusevska1, Rubens Jovanovik1, Neli Basheska1,
Sanja Trajkova1, Vesna Janevska1, Blagica Dukova1,
Magdalena Bogdanovska Todorovska1,
Hans Konrad Muller Hermelink2, Kennan Maclennan3,
Jacque Diebold4, Bharat Natwani5, Dennis Weisenburger6
1
Faculty of Medicine, University Ss. Cyril and Methodius, Skopje, Macedonia
Faculty of Medicine, University of Wuerzburg, Wuerzburg, Germany
Faculty of Medicine, University of Leeds, UK
4
Hospital Hotel de Diue, Faculty of Medicine, University of Paris, Paris, France
5
Cedars-Sinai Medical Center, Los Angeles, CA USA
6
City of Hope National Medical Center, Department of Pathology, Duarte, CA USA
2
3
The recognition of several new types of non-Hodgkin’s lymphoma (NHL) in recent
years has led to proposals for changing lymphoma classifications to the new WHO
classification of NHL-s. However, the clinical significance of the new entities and
the practical application of the new WHO Classification of malignant lymphomas in
Macedonia has not been done yet. The aim of the presentation was to present the
frequency of different malignant lymphomas entities and their main epidemiologic
characteristics. We have collected 222 cases from January 2009 to August 2010
with diagnosed malignant lymphoma. For histological diagnosis were used paraffin
tissue sections from nodal, extranodal tissues as well as bone marrow biopsies,
stained with H.E, Giemsa, PAS and immunohistochemically for panel of lymphocytic
monoclonal antibodies by PT-LINK immunoperoxidase technique. In the cases with
ambiguous morphology molecular studies were done at the Institute of pathology
in Wuerzburg. Clinical data were taken from the University Clinic of Hematology,
Medical faculty in Skopje. The analysis of the consensus working group showed that
the most frequent lymphomas in the Macedonian population were B cell lymphomas
mostly diffuse large B cell lymphomas and marginal zone malignant lymphoma with
nodal and extranodal localization. One of the main conclusions was that for the
histological diagnosis it is very important to have good clinical information for the
disease presentation.
Keywords: malignant lymphoma, classification, mmunohistochemical study,
Macedonian population
Keywords: lymphoma, immunohistochemical, Macedonian
[PP-LYMP-064]
Figure 1. First patient, cerebrospinal fluid, cytospin slide, immersion oil 100x.
BURKITT LYMPHOMA PRESENTING AS ILECOLIC
INTUSSUSCEPTION IN A CHILD
Sabah Boudjemaa1, Linda Dainese1, Julie Lemale2, Guy Leverger3,
Georges Audry4, Aurore Coulomb1
1
Department of Pathology, Armand Trousseau Hospital, Paris, France
Department of Gastroenterology, Armand Trousseau Hospital, Paris, France
Department of hemato-oncology, Armand Trousseau Hospital, Paris, France
4
Department of Surgery, Armand Trousseau Hospital, Paris, France
2
3
Background: Burkitt lymphoma represents 8-10% of all tumors in children under 15
years of age. Primary gastrointestinal lymphoma represents 1-4% of all gastrointestinal malignancies. Most cases in children involve the distal ileum or ileocecal region
with nonspecific symptoms, which can make diagnosis difficult. We present a case
presenting as ileocolic intussusception with clinical suspicion of atypical polyposis.
Case: A 13 year-old boy presented to the emergency room with intermittent abdominal pain, asthenia, anorexia, weight loss and sweats.
Figure 2. Second patient, cerebrospinal fluid, cytospin slide, immersion oil 50x.
On ultrasound, there was ileocecal and ileoileal intussusception confirmed by CT
scan. Once intussusception reduced, sessile masses of right colon, cecum and appendix were identified on follow-up ultrasound. A colonoscopy was performed 5
days later and showed several sessile colonic and intestinal polyps. Multiple endoscopic biopsies were performed. Meanwhile, the child presented a new episode
of intussusception with rectal hemorrhage leading to perform intestinal resectionanastomosis in emergency.
Results: Endoscopic colonic biopsies showed infiltration by a high grade lymphoma.
Tumor cells were medium-sized with a monotonous appearance. The cells appeared
cohesive with scant cytoplasm, round and uniform nuclei, 2 or 3 nucleoli and high
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Resected specimen consisting in 5 cm and 4 cm partial jejunectomy, 4 cm ilectomy,
showed intraluminal polypoid sessile masses measuring 3 to 4 cm. Two other polypoid tumors were identified in the appendix wall. All these lesions had the same
histological appearance of Burkitt Lymphoma.
The patient received chemotherapy according to LMB 2005 protocol and remains in
complete remission 3 years later.
Conclusion: Diagnosis of abdominal Burkitt Lymphoma may be challenging in children because of nonspecific clinical presentation. Rapidity of volumetric doubling of
this aggressive neoplasm may lead to an acute abdomen, mimicking other diseases,
such as polyposis with intussusception. Burkitt lymphoma should then be considered in differential diagnosis of acute abdomen in children.
Keywords: burkitt lymphoma, children, intussusception
Aims: To examine the expression of CCR4 in DLBCL, NOS cells and analyze the
relationship between the CCR4 expression and patients’ prognosis and, in addition,
Foxp3 expression in DLBCL, NOS cells.
Methods: Eighty lymph nodes from patients with DLBCL, NOS were immunostained
with antibodies against CCR4 and FoxP3. The relationship between the CCR4 expression and patients’ prognosis was analyzed using the Kaplan–Meier method and
Cox proportional hazards model.
Results: Of 80 patients aberrant CCR4 expression was not less than that in 10
(12.5%) cases (Figure 1). All CCR4-positive DLBCL cells were negative for FoxP3.
There was no significant difference in overall and progression-free survival (OS and
PFS) between CCR4-positive and CCR4-negative DLBCL, NOS patients (OS; P =
0.3236, PFS; P=0.6334, log-rank test) (Figure 2). CCR4-positive DLBCL, NOS may
not originate from FoxP3-positive Breg cells and therefore could not result in tumorinduced immunosuppression.
Conclusion: Mogamulizumab, a therapeutic antibody targeting CCR4 used for ATLL,
could be a new strategy for patients with CCR4-positive DLBCL, NOS.
[PP-LYMP-065]
Keywords: Diffuse large B-cell lymphoma not otherwise specified, CCR4, FoxP3
THE PREVALENCE OF EPSTEIN-BARR VIRUS-POSITIVE LYMPHOID
CELLS IN NASAL MUCOSA: AN EXTREMELY RARE EVENT
Sanghui Park
Department of Pathology, EWHA Womans University School of Medicine, Seoul, Korea
Background: The prevalence of EBV (Epstein-Barr virus)-positive lymphoid cells is
unknown. Because EBV is implicated in the etiology of extranodal NK/T-cell lymphoma, nasal type (nasal ENKL), the presence of EBV-positive lymphoid cells (EPLs)
in nasal mucosa specimens is expected. This study evaluated the presence of EBVpositive lymphoid cells in the nasal mucosa of 420 patients who had undergone
surgical resection of lesions of the nasal cavity due to nasal septal deviation, chronic
paranasal rhinosinusitis, chronic hypertrophic rhinosinusitis, nasal polyps, allergic
rhinitis, papillomas, and cysts.
On immunohistochemistry, tumor cells expressed CD20, CD79a, CD10 and BCL6.
Proliferation marker Ki67 was very high (100% of tumor cells). In situ hybridization
for EBV-encoded RNA (EBER) was negative, as BCL2. This morphology and immunophenotype were consistent with Burkitt Lymphoma.
inflammatory diseases. Foxp3 is the key transcription factor that is likely to play a
key role in controlling the expression of critical suppression-mediating molecules.
Recently, the presence of Foxp3+ B regulatory cells (Breg cells) was recently reported, and these cells appear to have a function similar to that of Foxp3+ Treg cells
and negatively regulate allergic diseases.
mitotic index. They molded to each other in a “mosaic tile” pattern, with starry sky
pattern due to numerous histiocytes.
Figure 1. Immunohistochemistry with anti-CCR4 antibody.
Methodology/Principal: Three representative 1.0-mm-diameter core biopsies were
taken from one paraffin-embedded donor tissue block per case and subsequently
arranged in new recipient paraffin blocks with a trephine. EBV in situ hybridization
study was performed to detect EPLs.
Results: None of the cases demonstrated EPLs.
Conclusions: The presence of EPLs in the nasal mucosa is an extremely rare event
in immunocompetent individuals. Therefore, the detection of EPLs in nasal biopsy
specimens should prompt the pathologist to perform further testing to exclude the
possibility of nasal ENKL.
Keywords: Epstein-Barr Virus; Nasal Mucosa
[PP-LYMP-066]
ABERRANT EXPRESSION OF CCR4 IN DIFFUSE LARGE B-CELL
LYMPHOMA, NOT OTHERWISE SPECIFIED
Motomu Tsuji1, Shoko Nakayama2, Taiji Yokote2, Yoshinobu Hirose1
1
Pathology, Osaka Medical College, Takatsuki city, Japan
Internal Medicine I, Osaka Medical College, Takatsuki city, Japan
2
Background: Diffuse large B-cell lymphoma, not otherwise specified (DLBCL, NOS)
is the largest category of aggressive lymphoma. CC chemokine receptor 4 (CCR4)
is an important chemokine receptor for regulating immune balance, and selectivity
expressed on Th2 and Forkhead box P3 (Foxp3) -positive regulatory T (Treg) cells,
but not in normal B-cells. CCR4 is expressed in about 80% of adult T-cell lymphoma/
leukemia (ATLL), and CCR4 is a poorer prognostic marker in ATLL. However, there
are no reports regarding the aberrant expression of CCR4 and the relationship to
patients’ prognosis in DLBCL, NOS. Previous studies on immune mechanisms have
mainly focused on the effector functions of immune responses during immunopathogenesis. Since the discovery of FoxP3 T regulatory cells (Treg cells), which
suppress antigen (Ag)-specific immune effector cells, negative immune regulation
has been noted to characterize the pathogenesis of autoimmunologic and allergic
Figure 2. Overall survival and progression free survival.
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111
th
[PP-LYMP-067]
IMMUNOGLOBULIN REARRANGEMENT ANALYSIS USING NEXT
GENERATION SEQUENCING: NEW INSIGHTS FROM MULTIPLE
LESIONS IN THE SAME PATIENT
Patricia Groenen1, Silke Appenzeller2, Christian Gillissen3,
Jos Rijntjes1, Annemiek Kastner Van Raaij1, Konnie Hebeda1,
Bastiaan Tops1, Loes Nissen4, Bas Dutilh2, Han Van Krieken1
1
Department of Pathology, Radboud University Nijmegen Medical Center, Nijmegen, The
Netherlands
2
Center for Molecular and Biomoleclar Informatics, Radboud Institute for Molecular Life Sciences,
Radboud University Nijmegen Medical Centre, Nijmegen, the Netherlands
3
Department of Genetics, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands
4
Department of Gastroenterology and Hepatology, Radboud University Nijmegen Medical Center,
Nijmegen, The Netherlands
For patients who have multiple lymphomas with discordant morphology and/or phenotype, it is relevant to determine whether there is one disseminated lymphoma or
two unrelated lymphomas. In this study we have used next generation sequencing
(NGS) to characterize the immunoglobulin heavy (IGH) gene V-D-J rearrangements
in multiple samples of two patients with iatrogenic immunodeficiency–associated
Epstein-Barr virus lymphoproliferative disorder (EBV-LPD), with ulcerative colitis as
underlying disease.
Next generation sequencing was performed by semiconductor sequencing on the
Ion Torrent-Personalised Genome Machine and IG-rearrangements were assessed
by the multiplex EuroClonality/BIOMED-2 IGH-FR3 primer set. Analysis of the NGS
data was done using the ClAnSort tool, a custom made analysis program which was
developed by group.
Evaluation of the rearrangement pattern involving assessment of the IGH (DJ and
VDJ) and IGK (VJ and KDE) rearrangements by fragment analysis (Figure 1, Figure
2), which is currently the golden standard for rearrangement detection, was not
able to either confirm or exclude clonal relationship. The IG sequences obtained by
NGS and subsequent data-analysis revealed undoubtedly the presence of the same
B-cell clone in patient C1 and two separate lymphomas in patient C2 (Figure 3). The
interpretation of the obtained IG-sequencing results was supported by the clinical
findings seen in these patients.
Our study demonstrates the diagnostic application of next generation sequencing
for IG-assessment for clinical decision making in patients with several simultaneous
or subsequent lymphoproliferations.
Keywords: Epstein-Barr virus, immunoglobulin rearrangement, next generation
sequencing
Figure 2. IGH-FR3 gene rearrangement patterns of sequential biopsies of two
patients with EBV-LPD, obtained by GeneScanning (A-D) and by NGS and visualized
as read length plots (E-H). Cases C1_A (A and E) and C1_B (B and F) are from
patient 1: the patterns A and E show a dominant clonal product in a polyclonal
background. The clonal product can hardly been seen in the patterns B and F,
obtained from the second colon biopsy of the patient. Cases and C2_A (C and G) and
C2_B (D and H) are from patient 2. The patterns C and G show a differently sized
clonal product compared to the patterns D and H, obtained from the second lesion
that was investigated from this patient.
Figure 3. Nucleotide sequence of the clonal IGH rearrangements of the sequential
biopsies of two patients with EBV-LPD The IGH-nucleotide sequences of the two
colon biopsies of patient 1 are identical and therefore fit to the presence of the same
B-cell clone. whereas the nucleotide sequences of the sequential biopsies in patient
2 are clearly different, which is in line with two separate lymphomas. Note: PCR
products were submitted to semiconducting sequencing on the Ion Torrent-PGM and
NGS data files were further processed using the tool ClAnSort.
[PP-LYMP-068]
CLASSICAL HODGKIN’S LYMPHOMA OCCURRING
SIMULTANEOUSLY WITH PLASMA CELL TYPE OF CASTLEMAN’S
DISEASE: CASE REPORT
Vesna Mihajlo Cemerikic Martinovic1, Neda Cedomir Drndarevic1,
Zorica Cvetkovic2, Tamara Ivan Martinovic3, Stefan Dojcin Dojcinov4
1
Department of Pathology, Beo-lab, Belgrade, Serbia
Department of Hematology, Clinical Hospital Center Zemun, Belgrade, Serbia
Institute for Histology and Embryology, School of Medicine, University of Belgrade, Belgrade,
Serbia
4
Department of Pathology, University Hospital of Wales, Cardiff, United Kingdom
2
Figure 1. IG gene rearrangement data of sequential biopsies of two patients with
EBV-LPD EBV-LPD patient 1: The entire pattern of IG gene rearrangements (IGHD-J, and V-D-J, IGK-V-J and IGK-DE rearrangements) may indicate the presence
of the same, clonally related processes in the two lesions. However, an accurate
comparison was not possible since the suboptimal DNA-quality of the first biopsy
did not allow detection of rearrangements in PCR targets (IGH-FR2 and IGK-DE)
that were informative in the second biopsy. EBV-LPD patient 2: The clonal IGKDE rearrangements have identical sizes, which would fit to a clonal relationship.
The IGH-FR3 rearrangements are different in size, which does not fit to a clonal
relationship. However, given the identical sized IGK-DE rearrangement, the IGHFR3 rearrangements might represent the same rearrangement with a long insertion
or deletion due to somatic hypermutation in one of the two, which would support
a common clonal origin of both biopsies. For these reasons, these results of the
clonality analysis were inconclusive to determine whether there is a clonal relationship
between the biopsies. # DNA quality, as measured using the gene control PCR that
amplifies fragments of 100, 200, 300 and 400 bp. The highest-sized band that was
amplified in this PCR is documented in the table, thus revealing a measure of the
DNA quality. * presence of suspect 161bp peak just outside the polyclonal “Gaussian”
curve Abreviations: C = clonal, P = polyclonal, PCB= polyclonal background, nsp = no
specific product, bp = base pair, nd= not determined.
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3
Background: Castleman’s disease (CD) is a rare atypical lymphoproliferative
disease associated with risk of developing lymphoma. The relationship between
Hodgkin’s lymphoma (HL) and plasma cell-type Castleman’s disease has been well
documented. There have been over 20 cases reported in the literature and nearly all
of them were either diagnosed concurrently.
Materials-Methods: We present a case of classical Hodgkin’s lymphoma co-occurring with plasma cell type of Castleman’s disease.
Results: A 24-year-old man presented with left axillary and supraclavicular lymph
node swelling and mild splenomegaly. The serum level of interleukin-6 (IL-6) was
elevated with mild anemia. C-reactive protein was positive. 2-microglobulin was
elevated as well as IgG. All other parameters were normal. HIV was negative. Biopsy
specimen from left axillary lymph node showed numerous polyclonal plasma cells
replacing the tissue. In this plasma cell infiltrate a few isolated large atypical mononuclear cells and typical Reed-Sternberg cells (RS) were found. These were strongly
positive for PAX5, CD30, CD15, MUM1 and fascin. RS cells were also strongly
Keywords: Castleman, Hodgkin’s lymphoma
PRIMARY PLASMABLASTIC LYMPHOMA OF THE BONE
MARROW IN A HIV NEGATIVE PATIENTE WITH CONCOMITANT
MYC AND BCL2 REARRANGEMENTS, CASE REPORT
Khalil Alnajjar, Rui Barreira, Jose Cabecadas, Antonio Almeida,
Susana Carvalho, Margarida Silveira, Maria Silva, Lara Neto
Instituto Portugues de Oncologi, Lisboa, Portugal
[PP-LYMP-069]
A CLINICOPATHOLOGICAL STUDY OF SPORADIC BURKITT
LYMPHOMA IN TAIWAN SHOWING 20% ASSOCIATION WITH
EBV
Bo Jung Chen1, Sheng Tsung Chang2, Shih Feng Weng3,
Wan Ting Huang4, Pei Yi Chu5, Pin Pen Hsieh6, Shih Sung Chuang7
1
Department of Pathology, Taipei Medical University Hospital, Taipei, Taiwan
Department of Pathology, Chi-Mei Medical Center, Tainan, Taiwan; Department of Nursing, National
Tainan Institute of Nursing, Tainan, Taiwan
3
Department of Medical Research, Chi-Mei Medical Center, Tainan, Taiwan
4
Department of Pathology, Kaohsiung Chang Gung Memorial Hospital-Kaohsiung Medical Center,
Kaohsiung, Taiwan; College of Medicine, Kaohsiung and Chang Gung University, Kaohsiung, Taiwan
5
School of Medicine, Fu-Jen Catholic University, New Taipei City, Taiwan; Department of Pathology,
St. Martin De Porres Hospital, Chiayi, Taiwan
6
Department of Pathology and Laboratory Medicine, Kaohsiung Veterans General Hospital,
Kaohsiung, Taiwan; Center for General Education, Yuh-Ing Junior College of Health Care and
Management, Kaohsiung, Taiwan
7
Department of Pathology, Chi-Mei Medical Center, Tainan, Taiwan; Taipei Medical University and
National Taiwan University, Taipei, Taiwan
2
Burkitt lymphoma (BL) is an aggressive B-cell lymphoma and is classified clinically into endemic, sporadic and immunodeficiency-associated variants. Nearly
all endemic and 25-40% of immunodeficiency-associated BLs are associated
with Epstein-Barr virus (EBV), while the association of EBV with sporadic cases
is variable in different geographic regions. To date, there is no large series of
sporadic BL from Taiwan. In this retrospective study, we collected 59 cases from
seven hospitals around Taiwan including 38 adult and 21 pediatric cases. We
performed histopathology review, immunohistochemistry including myc protein,
EBV in situ hybridization (EBER), and fluorescence in situ hybridization (FISH). We
analyzed the clinical data and correlated the clinicopathological variables with
outcome. There were 76% of cases with extranodal presentation; and 46% of
cases presented with abdominal tumors. Central nervous system (CNS) involvement and leukemic change at diagnosis or during the disease progression was
noted in 23% and 19% cases, respectively. Immunohistochemically, most cases
revealed typical immunophenotype of BL with CD10+, Bcl2-, Bcl6+, and >95%
Ki-67 proliferation index, and 88% (51/58) cases expressed myc protein diffusely
and intensively. EBER positive rate was 20% (12/59). The EBER-positive cases
more frequently presented as nodal and extra-abdominal than EBER-negative
cases, but there was no significant association with age or overall survival (OS).
MYC and IGH rearrangements were identified in 84% (46/55) and 78% (42/54)
cases, respectively. The concordance rate between myc protein expression and
MYC alteration by FISH was 76% (41/54). Excluding the cases with the prototypic
IGH/MYC translocation (87%; 40/46), we also found 2 (4%; 2/46) cases with IGL/
MYC, 1 (2%; 1/46) with IGK/MYC and 3 (7%; 3/46) with an unknown partner
other than IGH, IGK, or IGL. By multivariate analysis, we found that the OS was
significantly associated with age, CNS involvement, leukemic transformation, and
with or without CNS prophylaxis, but not with gender, EBV status, myc protein
expression, or MYC gene rearrangement, with or without receiving radiotherapy.
In conclusion, we reported the largest cohort of sporadic BLs from Taiwan and
characterized their clinicopathological and molecular features. We found that 20%
cases were associated with EBV, similar to the data from Japan and America, but
higher than that from China (5/43; 11%) and Korea (3/28; 11%). In our series,
there was no significant association of EBER-positivity with age, in contrast to
a recent Japanese study showing stronger EBV-association with patients older
than 50. We suggested that diffuse and intense myc protein expression might
serve as a surrogate marker for MYC translocation in laboratories where FISH
was not available.
Background: Plasmablastic lymphoma (PBL) was initially defined as an aggressive
B-cell lymphoma occurring in the oral cavity arising in the context of HIV infection. Cells have plasmablastic features, lack expression of B-cell markers but show
plasma cell differentiation antigens. Although most commonly observed in the oral
cavity of human immunodeficiency virus HIV-positive patients, it can rarely be observed at extra-oral sites and in HIV-negative patients. Additional molecular events
have been described including frequent IG/MYC translocations (49% in the largest
published series) and gains in multiple chromosomal loci with complex karyotypes.
Interestingly, rearrangements of both IGH genes were detected in 16% of cases
of another study with t(14;18) and t(11;14) respectively involved in conjunction
with a t(8;14) in two cases of HIV-related oral plasmablastic lymphomas. However,
no rearrangements of BCL6, MALT1 or PAX5 were detected in any plasmablastic
lymphoma.
Aims: To report a case of PBL confined to the bone marrow in an HIV-negative
patient and presence of a paraprotein (monoclonal  light chain), that has both
morphological and imunophenotypical characteristics of PBL, but having rearrangements both of MYC and t(14,18) BCL2-IGH genes.
Methods: A 77 year old HIV-negative female was referred to our institution with
fever, night sweats and weight loss. Physical examination showed no lesions in the
oral cavity, no lymph node enlargement nor organ infiltration. This was confirmed by
CT scan of the thorax, abdomen and pelvis. Bone marrow aspirate showed massive
infiltration by cells with “immunoblast-like” features. Bone marrow biopsy showed
a diffuse infiltration by large cells with a plasmablastic appearance and a “starry
sky” growth pattern; the cells
Conclusion: The diagnosis of classical HL with plasma cell variant of CD was made.
The source of elevated serum levels of IL-6 and for an abundant plasma cell reaction probably were RS cells. This type of HL can be easily misdiagnosed by pathologist due to the scarcity of RS cells and unusual CD back-ground.
[PP-LYMP-070]
positive for Epstein-Barr virus LMP1 and for Epstein-Barr virus encoded RNA by in
situ hybridization. EMA, LCA, CD20 and CD3 were negative in RS cells. HHV-8 was
not detected in RS cells and in surrounding tissue.
were CD20-, PAX5-, CD79a-, CD38+, CD138+, CD56-, MUM1+, CD30+ (focal),
CD3-, CD10-, EBER-, and expressed only lambda light chains. Ki67 was 80%. MYC
and BCL-2 gene rearrangements were detected by FISH. Although serum electrophoresis did not show monoclonal gammopathy, immunofixation detected monoclonal light chain.
Results: the morphological and genetic findings together with the very high proliferation rate and absence of clinical criteria for multiple myeloma lead to the diagnosis
of Plasmablastic lymphoma, with a genetic double hit.
Conclusion: To our knowledge the present case is the first report of PBL confined
to bone marrow with concomitant MYC and BCL-2 rearrangements. PBL is a rare
lymphoma and this case confirms the heterogeneity of its presentation (primary
bone marrow in a HIV-negative patient). The concomitant genetic involvement of
both MYC and BCL-2 suggests a probable aggressive course in this case.
Keywords: Plasmablastic lymphoma, MYC AND BCL-2 rearrangements
Keywords: Burkitt lymphoma; EBV; fluorescence in situ hybridization; MYC; Taiwan
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113
th
[PP-LYMP-071]
[PP-LYMP-073]
DBA.44 EXPRESSION IS A CONSISTENT FEATURE OF PRIMARY
SPLENIC MANTLE CELL LYMPHOMA WHEREAS IT IS NOT
DETECTED IN ITS NODAL COUNTERPART: ONTOGENETIC
IMPLICATIONS AND DIAGNOSTIC CONSIDERATIONS
EVALUATION OF CAPRIN1 PROTEIN EXPRESSION IN DIFFUSE
LARGE B CELL LYMPHOMAS
George Kanellis1, Anna Tasidou1, Niki Stavroyanni2, Evdoxia Pouliou1,
Themistoklis Karmiris3, Dimitrios Boutsis4, Konstantinos Tsionos5,
Achilles Anagnostopoulos2, Konstantinos Stamatopoulos6,
Theodora Papadaki1
1
Akdeniz University School of Medicine Pathology Department, Turkey
Akdeniz University School of Medicine Medical Biology and Genetics Department, Turkey
Bașkent University School of Medicine Pathology Department, Turkey
4
Akdeniz University School of Medicine Hematology Department, Turkey
2
3
1
Hematopathology Department, Evangelismos Hospital, Athens, Greece
Hematology Department and HCT unit, G. Papanicolaou Hospital, Thessaloniki, Greece
3
Hematology Department, Evangelismos Hospital, Athens, Greece
4
Hematology Department, Nautical Hospital, Athens, Greece
5
Hematology Department, 251 General Air Force Hospital, Athens, Greece
6
Institute of Applied Biosciences, Center for Technology and Research Hellas, Thessaloniki, Greece
2
Introduction: Mantle cell lymphoma (MCL) is a small B-cell lymphoma with mainly
nodal involvement generally following an aggressive clinical course. That notwithstanding, a relatively minor proportion of MCL displays more indolent clinical behavior. This particular subset is enriched for cases with primary splenic MCL and/or
primary involvement of the bone marrow (BM) in the absence of lymphadenopathy.
These cases are increasingly recognized as biologically distinctive and distinct from
nodal MCL.
Caprin1 is a protein that in humans is encoded by the cytoplasmic activation/proliferation-associated protein-1 gene located in 11p13 chromosome region. It has
been reported that Caprin 1 is associated with cell proliferation in various types of
cell lineages. The correlation of proliferation with increased expression of Caprin1
was also seen when levels of Caprin-1 in different tissues. The tissue with the highest level of Caprin-1 was the thymus, a site of continuous cellular proliferation. In
contrast, caprin-1 levels were low in tissues such as the kidney or muscle that have
a low proportion of dividing cells.
Material and Method: We researched whether Caprin 1 might be overexpressed
or not in Diffuse large B cell lymphomas (DLBCL) and its overexpression could be
correlated with clinicopathologic parameters (age, sex,). Caprin1 expression was
immunohistochemically evaluated in tissue microarrays of 99 Diffuse large B cell
lymphoma tissues.
Aim: The purpose of this study was to characterize in detail the immunohistological
profile of MCL cases with purely splenic involvement and BM infiltration, seeking for
additional insight into its ontogeny and relationship with nodal MCL.
Results: The expression of Caprin 1 was observed in 49 DLBCL cases (%49,4) (24
female, 25 male). However, 50 DLBCL were negative with Caprin1. Age range was
9-83 year.
Patients and Methods: We studied spleen samples (3/5) and BM biopsies (BMB)
(5/5) of five patients (4 males, 1 female) with MCL aged 58-73 years. No case had
lymphadenopathy; 3/5 cases had pronounced lymphocytosis. Methods: (a) BMB
and spleen morphology (H&E). (b) Immunohistochemistry for CD20, CD79, CD27,
CD5, CD23, bcl6, bcl2, CD10, cyclin D1, DBA44, CD3, SIgCIg, CD138, MUM1.
Conclusion: Caprin-1 overexpression might be correlated with the cellular proliferation potential of lymphomas. We revealed the expression profile of Caprin-1 in
human tumor tissues for the first time.
Results: (a) BM biopsy: lymphomatous infiltration of the bone marrow (20-75%)
mainly with interstitial and to a lesser extent diffuse, nodular and/or intrasinusoidal pattern of infiltration by predominately small lymphocytes. Immunophenotype
of B-cell lineage - 5/5: CD20+, CD79+, PAX5+, CD5+, CyclinD1+, DBA44+ (4/5
100% expression, 1/5 20% expression) CD3-, CD23-, bcl6, CD10-, MUM1- | 3/5
IgMD+ (b) Spleen: Nodular expansion of the white pulp with variable infiltration
of the red pulp by a monomorphous lymphomatous population consisting of small
and rarely medium sized cells. Immunophenotype: of B-cell lineage 3/3 CD20+,
CD79+, PAX5+, CD5+, CyclinD1+, DBA44+ (2/3 100% expression, 1/3 40% expression), CD3-, CD23-, bcl6-, CD10, 3/3 IgD+. In view of these findings, it is
relevant to mention that DBA.44 was not detected in any of 160 nodal MCL cases
currently evaluated in a parallel ongoing study from our group.
Conclusions: The herein reported morphological and immunohistochemical similarities between primary splenic MCL and splenic marginal zone lymphoma (SMZL)
prompt speculations about the potential effect of the splenic microenvironment in
shaping the malignant phenotype. From a diagnostic perspective, our study underscores the fact that extended immunohistochemical study of tissue samples from
spleen and BM of patients with splenomegaly is essential for establishing an accurate diagnosis, with obvious implications for patient management.
Keywords: DBA.44, splenic MCL, primary
[PP-LYMP-072]
WITHDRAWN
114
Bahar Akkaya1, Zafer Cetin2, Hampar Akkaya3, Mualla Ozcan1,
Sibel Berker Karauzum2, Aysen Timuragaoglu4
| EAHP - 2014
| 17-22 October 2014
To determine of importance of Caprin -1 overexpression new studies are necessary.
Keywords: Caprin-1, lymphoma
[PP-LYMP-074]
BIOMED-2 MULTIPLEX PCR-BASED B-CELL CLONALITY
ASSESSMENT IN TISSUE SAMPLES WITH SMALL B-CELL
LYMPHOPROLIFERATIONS
Mine Hekimgil1, Nazan Ozsan1, Duygu Aygunes2, Nur Selvi Gunel2,
Asli Tetik Vardarli2, Guray Saydam3, Filiz Vural3, Ayse Uysal3,
Murat Tombuloglu3
1
2
3
Department of Pathology, Ege University Faculty of Medicine, Bornova, 35100 İzmir, Turkey
Department of Medical Biology, Ege University Faculty of Medicine, Bornova, 35100 İzmir, Turkey
Department of Haematology, Ege University Faculty of Medicine, Bornova, 35100 İzmir, Turkey
Since lymphoid malignancies have a common clonal origin, the vast majority contain clonally rearranged immunoglobulin (Ig) and/or T-cell receptor (TCR) genes.
Between September 2007 and April 2014, a total of 141 cases were studied by
standardized BIOMED-2 multiplex polymerase chain reaction (PCR) heteroduplex
analysis for Ig heavy and light chain and/or TCR gene rearrangement. Of these, 37
cases with suspected small B-cell lymphoproliferations were evaluated for B-cell
clonality. The final diagnoses and results of clonality analysis are listed on Table 1.
The morphological findings leading to B-cell clonality analysis were small tissue
biopsies, lymphoid follicle formation, follicular colonization, heavy background of
T lymphocyte population, and problems of interpretation of Bcl-2 staining in marginal zone lymphoma; meanwhile prominent mantle zone formation, marginal zone
differentiation, heavy load of tingible body macrophages characteristic of reactive
germinal centers, predominantly diffuse pattern, Bcl-2 negativity, and high Ki67 index in follicular lymphoma. Factors complicating differential diagnoses of reactive
lymphoproliferations were small tissue biopsies, formation of MALT tissue, monocytoid B-cell hyperplasia, reactive inflammatory processes consisting of mixed Tand B-cell populations, and samples that still represent a clinical suspect, despite
the reactive morphology. Discordance between histopathological examination and
molecular studies was noted in two cases. The first one, although exhibiting a polyclonal gene rearrangement pattern, was diagnosed as cutaneous marginal zone
lymphoma on pathological examination and on clinical follow-up two years after the
diagnosis a rebiopsy was evaluated from the same site and diagnosed as relapse.
The bone marrow biopsy sample of another case of marginal zone lymphoma of the
Keywords: B-cell lymphoma, clonality analysis, PCR
Introduction: In most cases of systemic T-cell lymphoma the lesions are B-cell poor.
An exception is angioimmunoblastic T-cell lymphoma (AITL), often associated with
reactive follicles, immunoblasts and, in a few cases, with a large B-cell lymphoma.
Follicular helper T-cell lymphoma (FTCL) rare and like AITL has the cell of origin
the follicular helper T-cell. The lymphoma cells in FTCL populate lymphoid nodules,
sometimes leading to the appearance of progressive transformation of germinal
centers (PTGC). We hypothesize that similar to AITL, many FTCL are B-cell rich, making them difficult to diagnose and differentiate from B-cell lesions in the absence of
an elevated index of suspicion.
Table 1. The final diagnoses and results of B-cell clonality analysis
Primary
differential
diagnosis
Malignant
Suspicious for
malignancy
Benign
Final diagnosis
Number of
cases
Monoclonal
Polyclonal
MZL, extranodal
15
14
1
FL
6
6
-
PCFCL
1
1
-
MCL
1
1
-
MZL, nodal vs CLL
1
1
-
MZL, extranodal
1
1
-
DLBCL vs FL
1
1
-
CD
1
1
-
Reactive lymphoid
infiltration
10
1
9
37
27
10
TOTAL
MZL = Marginal Zone Lymphoma; FL = Follicular Lymphoma; PCFCL = Primary Cutaneous Follicle
Center Lymphoma; MCL = Mantle Cell Lymphoma; CLL = Chronic Lymphocytic Lymphoma; DLBCL =
Diffuse Large B-Cell Lymphoma; CD = Castleman’s Disease
[PP-LYMP-075]
CHRONIC LYMPHOCYTIC LEUKEMIA WITH ABERRANT
EXPRESSION OF CD57, DELETION OF 17P AND ADVANCED
DISEASE
Xiaodong Li1, Maryam Maryam Sharifian1, Evan Yung1, Ryan Phan2,
Faramarz Naeim2
1
Department of Pathology and Laboratory Medicine, Keck USC/LAC and USC Medical Center, Los
Angeles, USA
2
Department of Pathology and Laboratory Medicine, the University of California Irvine, USA
3
Department of Pathology and Laboratory Medicine, VA Greater Los Angeles Healthcare System,
Los Angeles, USA
Chronic lymphocytic leukemia (CLL) is the most common leukemia characterized
by co-expression of CD19, CD20, CD5, CD23 and restricted surface Ig light chain.
Although about 15 % of CLL cases have been reported to have an atypical immunophenotype including expression of T cell antigens, to our knowledge, concomitant
expression of CD57 on CLL cells has not been reported.
Here we report two cases of CLL with aberrant expression of CD57, an NK/T cell
associated antigen. In the first case, CD57 expression (80%) was detected during
the initial diagnosis when the patient presented with bleeding, anemia and systemic
lymphadenopathy. The second patient did not have CD57 detected until 8 years after
diagnosis, when the leukemia relapsed with pleural involvement. Deletion of 17p,
an ultra-high risk marker, was detected by Fluorescence in situ hybridization (FISH)
in both cases. Furthermore, the good prognostic factor, the IgV (H) mutational status
was not detected in either of these two cases.
In summary, two CLL patients with aberrant CD57 expression and del (17p) are reported. Severe symptoms together with the adverse prognostic factors, in these two
patients, are suggestive of a poor prognosis of CLL with aberrant CD57 expression.
Keywords: 1, CLL. 2, CD57
FOLLICULAR HELPER T-CELL LYMPHOMA: A B-CELL RICH
VARIANT OF T-CELL LYMPHOMA
Sory J. Ruiz, Claudiu V. Cotta
Robert J. Tomsich Pathology and Laboratory Medicine Institute, Cleveland Clinic, Cleveland (Ohio), USA
Material-Methods: 4 cases of FHTL were identified and immunostains for CD2,
CD3, CD4, CD5, CD7, CD8, CD10, CD20, CD21, CD30, BCL6, CXCL13, Ki67, PD-1
were performed. Epstein-Barr virus (EBV) infection status was investigated by insitu hybridization (EBER). B- and T-cell clonality studies using Biomed 2 sets of
primers were performed on DNA from the paraffin blocks. Flow cytometry was available in 3 cases.
Results: All patients were in the 7th decade of life, 3 female and 1 male. The sites
involved were cervical (3 cases) and axillary (1) lymph nodes. Staging information
was available in 2 cases: one was stage III, the other was stage IV. The pattern of the
infiltrate was nodular in all cases. Nodules were composed of small, mature B-cells
and were centered by intermediate-sized neoplastic T-cells. In one case residual,
reactive follicles were present. In 2 cases the neoplastic cells were negative for CD3,
but positive for the other T-cell markers. In 2 cases they were positive for CD10, PD1
was detected in 3 cases and CXCL13 in 2. The B-cells had no morphologic or immunophenotypic abnormalities, no clonal population was detected by flow cytometry.
There were no Reed-Sternberg cells. EBER was negative in all cases. There were
distorted networks of follicular dendritic cells in all cases, but none showed proliferating vessels. Clonally rearranged TCR gamma and beta chain genes were detected
in all cases, while the test for clonal IGH and IGK was negative.
[PP-LYMP-076]
parotid gland showed clear monoclonal products on PCR examination. Because the
latter may reflect true clonality of a minimal neoplastic population, a further detailed
pathological review was performed, and evaluated as pseudoclonality. In conclusion, it is recommended that in most cases with suspected B-cell lymphoproliferative disorders, differential diagnosis between small B-cell lymphomas and reactive
inflammatory processes should be mainly based on a combination of morphologic
and immunophenotypic studies, and in occasional complicated cases supplemented
with molecular clonality assessment, including close interaction between the clinician, pathologist and molecular biologist.
Conclusions: This study indicates that FTCL should be retained in the differential diagnosis of B-cell rich lesions, especially with nodular pattern. Cases with B-cell rich
nodules can be misdiagnosed as B-cell lymphomas or PTGC if the small clusters
of atypical T-cells are not noticed. A second conclusion supports the separation of
FTCL from AITL as a diagnostic entity, as they have different morphologic features,
in spite of a common cell of origin. In all cases FTCL was easily differentiated from
AITL based on the absence of EBV positive B-cells, the lack of vascular proliferation or of numerous reactive follicles. The neoplastic cells formed small nests, were
surrounded by unremarkable mantles of B-cells and were a small minority of the
specimen. In contrast to AITL, in FTCL there were no clonal B-cells by immunohistochemistry, flow cytometry or molecular studies.
Keywords: T-cell lymphoma, helper T-cell
[PP-LYMP-077]
WHAT IS THE IMPORTANT OF THE F-18 FDG PET/CT’S
FOR EVALUTATION OF LYMPHOMA INFILTRATION OF BONE
MARROW IN THE INITIAL STAGING OF LYMPHOMA
Funda Aydin1, Ozan Salim2, Orhan Kemal Yucel2, Deniz Ekinci2,
Tayfur Toptas3, Firat Gungor1, Adil Boz1, Akin Yildiz1, Bahar Akkaya4,
Levent Undar2
1
Akdeniz University School of Medicine, Nuclear Medicine Department, Turkey
Akdeniz University School of Medicine, Hematology Department, Turkey
3
Marmara University School of Medicine, Hematology Department, Turkey
4
Akdeniz University School of Medicine, Pathology Department, Turkey
2
Purpose: The evaluation of bone marrow infiltration (BMI) is of crucial importance
in the staging of lymphoma. Although bone marrow biopsy (BMB) is the reference
method for the evaluation of BMI, it has limitations (e.g taken from a single field,
invasiveness). The aim of this study was to assess the performance of 2-deoxy2-[18F]fluoro-D-glucose (18F-FDG) PET/CT against bone marrow biopsy (BMB) in
the initial diagnosis of bone marrow infiltration (BMI) in patients with Hodgkin’s
Lymphoma (HL) and Non-Hodgkin’s Lymphoma (NHL), retrospectively.
| 17-22 October 2014
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115
Results: For all cases, in visual assessment; in the evaluation of lymphoma infiltration of BM, PET / CT’s sensitivity 31%, specificity 85%, positive predictive value
(POV) 39% and negative predictive value (NOV) 79% have been identified. Visual
assessment in HL patients the sensitivity 80%, specificity 78%, POV 24% and NOV
98%; in NHL patients sensitivity 24%, specificity 90%, POV 56% and NOV 70% were
reported, respectively.
Figure 1. Histologic features of primary cutaneous EBVLPDs. This case shows
polymorphous background infiltrate with a mixture of small lymphocytes, plasma
cells, histiocytes, epithelioid cells, and immunoblasts. Tumor cells are positive for
EBER.
In semi-quantitative evaluation; when the SUVmax was accepted as >= 4 in the HL
cases, the sensitivity 80%, the specificity 68% were found respectively. When the
SUVmax was accepted as >=3,2 in the NHL cases the sensitivity 65%, the specificity 58% were reported, respectively.
Conclusion: PET / CT imaging is more valuable than microscopic examination of the
BM for the staging of the lymphoma. PET/CT detects more BM involvement in HL
and NHL compared with BMB. Its good concordance with BMB makes it a complementary technique, as it helps select the biopsy site in cases.
Keywords: lymphoma, (18F-FDG) PET/CT
[PP-LYMP-078]
th
Material and Methods: A total of 172 patients (50 women, 122 men; 64 HL, 108
NHL) referred to Akdeniz University School of Medicine, Pathology Departments between July 2009 and December 2013, for investigated BMI, were included in the
study. All patients were examined by whole-body 18F-FDG PET/CT scan for initial
staging. Evaluation of BM with PET/CT scan was done visually and semi-quantitatively. The maximum standard uptake value (SUVmax) was used as quantitative
parameters in the evaluation of FDG PET. (18)F-FDG PET/CT was considered positive for BMI in cases of uni-or multifocal bone marrow (18)F-FDG uptake that could
not be explained by benign findings on the underlying CT image or history. A final
diagnosis of BMI was considered if the BMB was positive.
THE CLINICOPATHOLOGICAL FEATURES OF PRIMARY
CUTANEOUS EBVLPDS: A COMPARATIVE ANALYSIS WITH
SYSTEMIC AGE-RELATED EBVLPDS WITH CUTANEOUS
INVOLVEMENT
Naoko Asano1, Masahiro Hagiwara2, Tomohiro Kinoshita4,
Shigeo Nakamura3
Figure 2. Overall survival. Overall survival of the AR-EBVLPD group was
significantly lower than that of the primary cutaneous EBVLPD group (P = 0.0077).
[PP-LYMP-079]
1
Department of Clinical Laboratory, Nagoya University Hospital, Nagoya, Japan; Department of
Molecular Diagnostics, Nagano Prefectural Suzaka Hospital, Suzaka, Japan
2
Department of Dermatology, Nagoya Graduate School of Medicine, Nagoya, Japan
3
Department of Pathology and Clinical Laboratories, Nagoya University Hospital, Nagoya, Japan
4
Department of Hematology and Cell therapy, Aichi Cancer Center, Nagoya, Japan
Epstein Barr virus-positive B-cell lymphoproliferative disorders (EBVLPDs) develop
as a consequence of various types of immunosuppression and comprise a heterogeneous group of malignancies with highly variable forms of pathological features.
In the 2008 WHO classification, age-related-EBVLPD (AR-EBVLPDs) has been listed
as EBV-positive DLBCL of the elderly without any known underlying immunosuppression or history of lymphoma. The main lesion of extranodal involvement in
AR-EBVLPDs, in order of incidence, is skin, lung, pleural effusion, stomach, and
tonsil. A comparison between EBV-positive and EBV-negative groups showed that
the incidence of cutaneous involvement was significantly higher in patients with AREBVLPDs than in those with EBV-negative DLBCLs. To further characterize primary
cutaneous EBVLPDs, we document the clinicopathological profiles of 9 patients with
primary cutaneous EBVLPDs (4 men and 5 women; median age, 73 years) and compare the features of primary cutaneous EBVLPDs with advanced stage presentations of AR-EBVLPD with secondary cutaneous involvement. In 4 primary cutaneous
EBVLPDs cases, immunosuppression resulted from methotrexate (MTX) treatment
for rheumatoid arthritis (RA), while in 5 other cases it resulted from age-related
immunosenescence. Four patients presented with isolated, sharply circumscribed
ulcers involving the skin, which were classified as EBV-positive mucocutaneous
ulcers (EBVMCUs). Lesions were histologically characterized by a polymorphous
infiltrate of inflammatory cells and atypical large B-cell blasts. The atypical cells
had a prototypic immunophenotype of CD20+ CD15- CD30+ EBER+. Our 9 primary
cutaneous EBVLPDs cases showed significantly milder clinical symptoms and better prognosis than AR-EBVLPD with secondary cutaneous involvement. The conservative management in primary cutaneous EBVLPDs patients is recommended, as
shown by the regression after discontinuation of MTX and a waxing and waning of
the clinical course.
Keywords: EBV, B-cell lymphoproliferative disorders, cutaneous ulcer
116
| EAHP - 2014
| 17-22 October 2014
IMMUNOSUPPRESSIVE RELATED INDOLENT CYTOTOXIC
T-CELL LYMPHOPROLIFERATIVE DISORDER OF THE GASTROINTESTINAL TRACT IN A PATIENT WITH CROHN’S COLITIS
Philippe Trougouboff
Tissue Diagnosis and Cancer Research Institute, Emek Medical Center, Afula, Israel
Inhibitors of tumor necrosis factor (TNF)-alpha are increasingly used to treat Crohn’s
disease which is not responsive to conventional treatment. Unfortunately lymphomas, among them Hepatosplenic T-cell lymphoma (HSTCL), are more and more
recognized complication of TNF-alpha inhibition treatment for autoimmune and inflammatory diseases. Here we describe the case of a 27 years old patient treated
with Adalimumab (Humira) for resistant Crohn’s disease, who developed a primary
intestinal CD8+ cytotoxic T-cell lymphoproliferative disease (TCLPD) three months
after the initiation of the immunosuppressive treatment. This TCLPD mimics T-cell
lymphoma of the gastrointestinal tract, an aggressive disease associated with a dire
prognosis. Biopsies of the colon showed foci of inflammation, rich in plasma cells
and eosinophils, many crypts with reactive changes infiltrated with neutrophils, and
a characteristic picture of focal active Inflammatory Bowel Disease (IBD). These foci
were associated with a dense infiltrate of small lymphocytes. Apart from these foci
of active IBD, the other biopsies from the colonic mucosa were normal. The small
lymphocytes in the aggregates were nearly all stained for CD2, CD3, CD5, CD7,
CD8, with a few CD4 positive T-cells and small groups of CD20 positive B-cells. The
CD4/CD8 ratio was inversed (1/4). The CD8 positive T-cells were TIA-1 positive and
negative for Granzyme-B, CD56, and CD57. A PCR study for T- cell clonality revealed
a monoclonal population positive for TCRB and negative for TCRG and TCRD, consistent for -T-cells, allowing for the exclusion of a HSTCL.
TNF-alpha inhibitor treatment was withdrawn and a “wait and see” follow up was
adopted, without any cytotoxic treatment. The subsequent endoscopic biopsies at
three, six months and 1 year after the original diagnosis showed a gradual restoration of the CD4 and CD20 population and a regression of the CD8 positive T-cell
infiltrates.
INTEGRATING CLINICAL AND PATHOLOGICAL FEATURES
IN RISK STRATIFICATION AND THERAPEUTIC DECISION
MAKING IN PATIENTS WITH EPSTEIN-BARR VIRUS-RELATED
LYMPHOPROLIFERATIVE DISORDERS; A ROLE FOR IG
CLONALITY STATUS
Patricia Groenen1, Jos Rijntjes1, Wendy Stevens2,
Lakshmi Venkatraman3, Mark Catherwood3, Lisette Van De Laar1,
Moniek Craenmehr1, Hongxiang Liu4, Hesham Eldaly5,
Marieke Van Rijn2, Han Van Krieken1, Walter Van Der Velden2
1
Department of Pathology, Radboud University Medical Centre, Nijmegen, the Netherlands
Department of Hematology, Radboud University Medical Centre, Nijmegen, the Netherlands
3
Haematology Department, Belfast City Hospital, Belfast, United Kingdom
4
Molecular Malignancy Laboratory and Department of Histopathology, Addenbrooke’s HospitalCambridge University Hospitals, Cambridge, United Kingdom
5
Haematology Department, Addenbrooke’s Hospital-Cambridge University Hospitals, Cambridge,
United Kingdom
2
Background: Retrospective analyses have revealed risk factors that predict outcome
in patients with EBV-related lymphoproliferative disorders (EBV-LPD). However, pretreatment risk stratification that can be used to guide therapeutic decisions remains
difficult and algorithms are lacking.
Methods: 62 patients with an EBV positive PTLD (41 hematopoietic stem cell transplantation and 21 solid organ transplantation) and 24 with iatrogenic EBV-LPD
were included. Clinical and pathological data were collected. Clonality testing involving IGH and IGK gene rearrangements were assessed by the BIOMED-2 PCRs.
Morphology was examined by experienced haematopathologists.
Figure 1. Original biopsy. First colonic endoscopic biopsy at the time of original
diagnosis. Dense cytotoxic T-cell lymphoid infiltrate in the lamina propria. HxE, CD3,
CD8, CD4, TIA-1, CD20. Original magnification x100.
Results: Evaluation of morphology, clonality, clinical stage and mortality showed
a mortality rate of 35% in high-stage patients that displayed polymorphic/reactive morphology and IG monoclonality (Figure 1). In iatrogenic EBV-LPD Ann-Arbor
staging seemed most predictive for outcome. Stage I disease (group 1) was effectively cured in most patients by only modifying immunosuppressive therapy regardless the clonality status. More advanced stages, II-IV (group 2), mostly being
monoclonal, required treatment with rituximab alone or combined with chemotherapy (R/R-chemo), but still a mortality rate of 25% was encountered (Figure 2).
In patients with monomorphic PTLD (group 3) mortality was considerable despite
the use of R/R-chemo, being 38,5% (Figure 3). 90% of these patients had monoclonal disease. In patients with reactive/polymorphic PTLD the clonality status
seemed important. Monoclonality was associated with an unfavorable outcome
(mortality rate of 33%; group 4), which is comparable to the monomorphic PTLD.
However, a considerable number of deaths (5/11) were caused by undertreatment
probably resulting from inadequate risk assessment based on morphology alone.
In contrast, reactive/polymorphic PTLD that proved to be polyclonal had a good
outcome (group 5; Figure 3).
Keywords: lymphoproliferative, immunosuppressive, gastrointestinal tract
[PP-LYMP-080]
With the increased use of immunosuppressive drugs for autoimmune and chronic
inflammatory diseases such cases of indolent TCLPD should be recognized to avoid
a misdiagnosis and inadequate aggressive treatment for the patients.
Conclusion: IG Clonality status can be used to determine which patients with reactive/polymorphic PTLD can be managed by just reducing immunosuppressants
(polyclonal) or require more intensive therapy (monoclonal).
Keywords: IG Clonality, EBV, lymphoproliferative disorder
Figure 1. Clinicopathological features and outcome of patients with EBV-LPD.
Figure 2. CD4 and CD8 immunostains at diagnosis, 3 months and 12 months.
The CD8-positive T-cell infiltrate in the lamina propria progressively regress after
TNF-alpha inhibitor withdrawn. Original magnification x40.
| 17-22 October 2014
| EAHP - 2014
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117
th
[PP-LYMP-081]
CAUSATIVE AGENTS OF KIKUCHI-FUJIMOTO DISEASE
(HISTIOCYTIC NECROTIZING LYMPHADENITIS):
A META-ANALYSIS
Yosep Chong, Tae Jung Kim, Eun Jung Lee, Chang Suk Kang
The Catholic University of Korea College of Medicine, Yeouido St. Mary’s Hospital, Department of
Hospital Pathology, Seoul, Republic of Korea
Background: Kikuchi-Fujimoto disease (KFD) is a self-limiting disorder characterized by histiocytic necrotizing lymphadenitis in the cervical lymph nodes of young
women. Although an infectious etiology has been postulated, a definitive causative
agent has not been identified. The few dozens of published studies are limited by
small sample size and poorly structured study designs.
Figure 2. Flowchart of the clinicopathological features of patients with EBVpositive iatrogenic immunodeficiency-associated LPD. The clinical stage is the
risk determining factor for the patients treatment and outcome. The number of
patients that received treatment are indicated in the green boxes. Single-agent
rituximab (R) or rituximab with chemo (R-chemo). Outcome is expressed as
mortality rate and indicated in the red boxes. C= Monoclonality of immunoglobulin
gene rearrangements, nonC= polyclonal or oligoclonal immunoglobulin gene
rearrangements.
Materials and Methods: To evaluate the association of each infectious agent to KFD
that has been studied, we performed metaanalysis using major electronic database
(MEDLINE(pubmed), Cochrane library (Central), Embase, Web of Science, NML gateway, LILACS, and Google Scholar). Cross-sectional studies on the positivity of each
agent in clinicopathologically diagnosed KFD and matched controls by polymerase
chain reaction (PCR) or in situ hybridization (ISH) were carefully retrieved. The included infectious agents were herpes simplex virus (HSV) type 1, 2, varicella-zoaster
virus, cytomegalovirus, Epstein-Barr virus (EBV), human herpesvirus (HHV) 6, 7, 8,
parvovirus B19, human papilloma virus, hepatitis B virus, human T-lymphotropic
virus 1, Brucella, and Bartonella henselae. After an exclusion process of 2491 studies, six, three, four, two, two, and three studies on EBV-PCR, EBV-ISH, HHV6-PCR,
HHV8-PCR, parvovirus B19-PCR and HHV7-PCR, respectively, were suitable for
meta-analysis.
Results: None of them revealed a significant association with KFD compared to the
controls.
Conclusions: Our study verified that none of viral agents that were most commonly
found to be related with human diseases was associated with KFD more than controls. More studies focusing on the histologic features of KFD are necessary to identify the causative agent.
Keywords: Kikuchi-Fujimoto disease, histiocytic necrotizing lymphadenitis,
meta-analysis
[PP-LYMP-082]
SIP-F1: CHARACTERIZATION OF A NEWLY ESTABLISHED GRAY
ZONE LYMPHOMA CELL LINE
Benjamin Rengstl, Frederike Schmid, Martin Leo Hansmann,
Sebastian Newrzela
Goethe-University Clinic Frankfurt, Dr. Senckenberg Institute of Pathology
Figure 3. Flowchart of the clinicopathological features of patients with EBVpositive PTLD. Patients with PTLD with polyclonal or reactive morphology and
monoclonal IG gene rearrangements (group 4) have a high mortality and need more
intensive therapy. The number of patients that received therapy are indicated in
the green boxes. R / R-chemo: Single-agent rituximab (R) or rituximab with chemo
(R-chemo). Outcome is expressed as mortality rate and indicated in the red boxes.
Not monoclonal = polyclonal or oligoclonal immunoglobulin gene rearrangements.
118
| EAHP - 2014
| 17-22 October 2014
Lymphoid cancers are mainly classified according to their clinical presentation,
morphology, immunology and molecular genetic features. Diagnosis of classical
Hodgkin´s lymphoma (cHL) is mainly based on pathognomonic multinucleated, giant
Hodgkin and Reed/Sternberg cells (HRS). However, in the group of HRS cell-lacking
non-Hodgkin lymphomas (NHL), diffuse large B-cell lymphoma (DLBCL) can resemble typical features of cHL. In some cases, a diagnostic pitfall exists at the interface
of cHL and NHL, so that, morphological overlaps and missing clear-cut diagnostic
criteria complicate classification of these so-called gray zone lymphomas (GZL). In
a recent study, we analyzed a newly established GZL cell line derived from a lymph
node biopsy of a male 20-year-old lymphoma patient. Histological sections of the
primary GZL were characterized by striking pleomorphism of in part multinuclear
blast infiltrates and areas of necrosis. Immunohistochemistry revealed tumor cell
positivity for the markers CD15, CD30 and CD20. Subsequently, cell suspensions
were prepared from tumor tissue and after several weeks of passaging and under
normal cell culture conditions, we were able to establish a new GZL cell line (SIP-F1).
SIP-F1 presented clonal Ig gene rearrangement and flow cytometry analysis showed
expression of B-cell markers CD20 and CD19. Moreover, 50-60% of the established
cell line displayed HL marker CD30. Interestingly, but yet quite surprisingly, first
cytogenetic analysis revealed a normal karyotype for SIP-F1. For closer characterization and further differentiation of the new cell line, we performed gene expression
profiling (GEP) and compared several prominent B-cell lymphoma lines. However,
we were not able to classify SIP-F1, as the GEP of the cell line did not cluster with
any of the compared lymphoma entities. Nevertheless, we observed tumor inducing
growth of SIP-F1 in an immunodeficient mouse model, morphologically resembling
[PP-LYMP-083]
EVALUATING EXPRESSION OF DOWNSTREAM OF KINASE (DOK)
ADAPTOR MOLECULES IN LYMPHORETICULAR TISSUES WITH A
FOCUS ON THEIR USE AS NOVEL MARKERS FOR MAST CELLS
AND RELATED NEOPLASMS
Hasan Rizvi1, Laura Casey2, James Wilton3, Ayse U. Akarca3,
Vishvesh Shende3, Teresa Marafioti2
1
Barts Health NHS Trust
2
UCLH NHS Foundation Trust
3
University College London
The Downstream of Kinase/docking (Dok) family of proteins - currently with seven
members, Dok 1-7 - are adaptor proteins that function as substrates of both receptor and non-receptor tyrosine kinase. Though they share structural homology they
vary in function and are currently divided into two groups, group A with Dok1-3 that
are negative regulators distributed predominantly in haematolymphoid tissues and
group B with Dok4-7 that are distributed mainly in non-haematolymphoid tissues.
(Dok-4 - epithelial tissues +/- nerves, Dok5-6 - neural tissue and Dok-7 - muscle).
In the current study, we focussed on Dok1-3 and Dok-5 based on their tissue distribution and due to lack of availability of suitable antibodies for use in formalin fixed
paraffin embedded (FFPE) tissue for the other Dok proteins. The antibodies used for
this study included two mouse monoclonals Dok1 (IgG1, clone A3) and Dok2 (IgG1,
clone E10) a rabbit polyclonal Dok3 (H206) all from Santa Cruz Biotechnology, CA,
USA; and a goat polyclonal against Dok5 (B0023) from Everest Biotech (Oxford, UK).
All antibodies were tested at a range of dilutions and the optimal concentration
showed absence of background and selective cellular labelling. Conventional immunohistochemistry was performed on routine sections (3 micron thickness) from
FFPE tissue blocks of normal and neoplastic haematolymphoid tissues selected
from the histopathology archives.
Table 1. Distribution of Dok1-3 and Dok-5 in normal haematolymphoid
tissues
TONSIL/LYMPH NODE
-Lymphoid follicles
--Germinal centers
--Mantle zones
-Monocytoid B-cells (1)
-Plasma cells
-T cells
SPLEEN
-B cell areas
--Marginal zones
--Mantle zones
-T cell areas
-Red pulp
THYMUS
-Cortex
-Medulla
BONE MARROW
-Erythroid
-Granulocytic
-Mast cells
-Megas
-Plasma cells
Dok1
Dok2
Dok3
Dok5
Negative
Negative
Negative
Positive
Negative
Negative (3)
Negative
Negative
Negative
Positive
Positive (5)
Positive
Positive
Weak
Negative (6)
Negative
Negative
Negative
Negative
Negative
Negative
Negative
Negative
Negative
Negative
Negative
Positive
Negative
Positive
Positive
Negative
Negative
Negative
Negative
Negative
Negative
Negative
Negative
Weak (4)
Positive
Negative
Rare
Negative
Negative
Negative
Negative
Negative
Negative
Positive (2)
Negative
Negative
Negative
Weak
Negative
Negative
Positive
Negative
Positive
Negative
Negative
Negative
Positive (7)
Negative
Negative
(1) This cell population was identified in lymph nodes reacting to toxoplasmosis. (2) Dok1 staining was restricted to plasma
cells in all of the normal hematopoietic tissues evaluated. (3) Dok2 stained histiocytes weakly in hematopoietic tissues. (4)
Scattered weak staining was also seen in cortical thymocytes by Dok2. (5) Dok3 was weakly positive in histiocytes. (6) Dok3
was strongly expressed in a few lymphocytes scattered in interfollicular areas, of which a subset showed morphology of
activated lymphocytes. (7) Dok5 staining was restricted to mast cells in all of the normal hematopoietic tissues evaluated.
Dok1 (clone A3) expression is reported. Another antibody was also evaluated with similar staining patterns (clone 36), but
weaker expression. Dok2 (clone E-10) expression is reported. Another antibody (clone 28) was also evaluated with similar
staining patterns, but weaker expression.
Keywords: Gray zone lymphoma
the primary tumor. Histological analysis of tumor tissue demonstrated large tumor
cells, including lacunar, R-S like cells growing in sheets. In future studies of SIP-F1
we want to determine the EBV-status of the cell line and perform next generation
sequencing. In conclusion, considering the paucity in prospective differentiation of
GZL cases, their distinct clinical behavior and treatment, we think that SIP-F1 could
be a helpful tool to study morphological and molecular features of GZL and to better
understand the biology of these unusual cases.
Table 2. Immunohistochemical staining of downstream of Kinase (DoK)
proteins in human haematolymphoid neoplasms
The immunohistochemical results are summarised in Tables 1-3 and outlined below:
Normal haematolymphoid tissues:
Dok-1: Mainly in mature plasma cells;
Dok-2: nearly exclusive expression in T-lymphocytes in the tonsil, lymph node,
spleen and thymus (strong staining in the medulla, weaker in the cortex);
Dok-3: Positive in B-cells with weak plasma cell staining. Positive in BM granulocytes and megakaryocytes;
Dok-5: Expression limited to bone marrow mast cells.
Mast cell neoplasms:
Dok-1: Weak nucleolar staining in 2/8 cases of Cutaneous mastocytosis (CM); Dok2: Variable weak nuclear +/- cytoplasmic staining in 5/9 Cutaneous Mastocytosis,
3/7 Systemic mastocytosis and 1/1 Mast Cell Leukaemia (MCL).
Dok-3: None.
Dok-5: 100% in Mastocytosis (8/8 Cutaneous Mastocytosis, 8/8 Systemic
Mastocytosis and 1/1 Mast Cell Leukemia. Negative in Non-Hodgkin and Hodgkin
Lymphomas.
Table 3. Comparison of Dok-5 with commonly used mast cell markers
All investigated DOK molecules were negative in myeloid neoplasms.
Our study highlights the immunodiagnostic utility of Dok-5 in identifying mast
cells in normal tissues and in the diagnosis of mast cell related neoplasms. Also,
Dok-5 is negative in myeloid neoplasms compared to some other mast cells markers (See Table 3) which may give it a diagnostic edge in bone marrow evaluation.
Furthermore, other Dok proteins show that their expression is confined to distinct
haematolymphoid tissues and neoplasms (see Table 2).
Keywords: Downstream of Kinase (DoK) proteins, Diagnostic immunohistochemistry, Mast cell neoplasms
| 17-22 October 2014
| EAHP - 2014
|
119
th
[PP-LYMP-084]
MIB1 IN CLL/SLL CORRELATES WITH INFILTRATING CD3
POSITIVE T CELLS AND ATYPICAL NUCLEAR MORPHOLOGY
Tanuja Shet, Kiran Ghodke, Sridhar Epari, Sumeet Gujral,
Hari Menon
Tata Memorial Hospital, Parel, Mumbai, 400012, India
Background: There is no data on prognostic significance of MIB1 in SLL/CLL. This
study evaluated 64 patients included 50 males and 14 females with a view to
evaluate adverse prognostic impact of MIb1. A CD38 and ZAP70 staining was also
performed on the nodes. CD3 immunohistochemistry was used to quantify tumor
infiltrating lymphocytes in these nodes.
sites other than the spleen demonstrated dim to negative staining for both CD200
(1/5 (20%) dim, 4/5 (80%) negative) and CD1d (2/5 (40%) dim, 3/5 (60%) negative);
these cases were also negative for CD103, but showed positivity for CD25 and
CD11c in 2/5 cases. No positive staining for CD200 was seen in any case of SMZL
or marginal zone lymphoma involving sites other than the spleen.
Conclusions: Flow cytometric analysis of CD200 and CD1d, along with CD25,
CD103, and CD11c, reliably distinguishes lymphoplasmacytic lymphoma, hairy cell
leukemia, hairy cell leukemia-variant, marginal zone lymphoma, and splenic marginal zone lymphoma, and should be incorporated into flow cytometric panels used
in the diagnosis of CD10-negative, CD5-negative B-LPDs.
Keywords: Hairy cell leukemia, lympoplasmacytic lymphoma, marginal zone
lymphoma
Results: Age group ranged from 33 to 93 years with both mean and median of 60
years. Rai Stage in 29 patients was Stage I, Stage II in 16 patients, Stage III in 6
patients and Stage IV in 13 patients. Seventeen tumors had atypical morphology
with cleaved, often nucleolated pleomorphic cells. MIB1 ranged from 10 to 70%
with a mean of 26%. MIB1 correlated significantly with increased presence of CD3
staining T cells within the nodes( p value – 0.0001). MIb1 also correlated with atypical nuclear morphology (p value – 0.01). MIB1 staining also marginally correlated
with ZAP70 staining in the lymph node (p value 0.054). MIb1 did not correlate with
Rai stage or CD38. Rai Stage, CD3 percentage ( p – 0.307) ZAP70( p 0.794), CD38(
p – 0.678), increase in growth centers( p value-0.587) and MIb1( p value 0.282) did
not impact overall survival of patients.
Conclusion: As opposed to other low grade lymphomas tumor, infiltrating T cells
may add to the increased detection on MIb1 in SLL/CLL and hence has lesser value
in prognostication of this lymphoma.
Keywords: Small lymphocytic lymphoma, chronic lymphatic leukemia, MIB1
Figure 1. CD1d and CD200 expression paterns in CD10-/CD5- mature B-cell
lymphomas.
Table 1. Antigenic profiles of CD10-/CD5- mature B-cell lymphomas
[PP-LYMP-085]
FLOW CYTOMETRIC PATTERNS OF CD200 AND CD1D
EXPRESSION DISTINGUISH CD10-NEGATIVE, CD5-NEGATIVE
MATURE B-CELL LYMPHOPROLIFERATIVE DISORDERS
Emily F. Mason, Olga Pozdnyakova, Betty Li, David M. Dorfman
Department of Pathology, Brigham and Women’s Hospital and Harvard Medical School, Boston,
MA, USA
Objectives: The importance of distinguishing mature B-cell lymphoproliferative
disorders (B-LPDs) is highlighted by the distinct treatments used for and varying
prognoses seen in association with these different diseases. Immunophenotyping
allows for accurate and efficient differentiation of many B-LPDs. Due to overlapping
immunophenotypes and antigenic variation, panels including multiple flow cytometric markers are often more effective in diagnosing B-LPDs than is any single marker
used alone. Recently, we showed that CD200 is highly expressed in hairy cell leukemia, but not in marginal zone lymphoma, lymphoplasmacytic lymphoma, or hairy
cell leukemia-variant. Here, we have assessed the usefulness of a flow cytometric
panel combining CD200 with CD1d, CD25, CD103, and CD11c to distinguish among
CD10-negative, CD5-negative B-LPDs.
Methods: We analyzed 15 cases of lymphoplasmacytic lymphoma (LPL), 11 cases
of hairy cell leukemia (HCL), 1 case of hairy cell leukemia-variant (HCL-v), 5 cases
of marginal zone lymphoma (MZL), and 4 cases of splenic marginal zone lymphoma
(SMZL) for expression of CD200-PerCP-Cy5.5, CD1d-PE, CD11c-APC, CD25-PECy7, CD103-FITC, and CD19-APC-Cy7 (all from BD Biosciences) by flow cytometry.
The samples analyzed included 20 bone marrow, 13 peripheral blood, 1 spleen, and
2 lymph node specimens. Expression of CD200 and CD1d was scored as negative,
dim or positive based on fluorescence intensity.
Results: Distinct patterns of CD200 and CD1d expression were seen in the examined B-LPDs (Table and Figure). LPL demonstrated dim or negative staining for
both CD200 (7/15 (47%) dim, 8/15 (53%) negative) and CD1d (11/15 (73%) dim,
4/15 (27%) negative) and was also negative or dimly positive for CD25, CD103, and
CD11c. No positive staining for CD200 or CD1d was seen in any case of LPL. HCL
demonstrated positive staining for both CD200 and CD1d in 11/11 (100%) cases
and stained positively for CD25, CD103, and CD11c. In contrast, the one case of
HCL-v examined was negative for both CD200 and CD1d as well as for CD25 and
CD103, but was positive for CD11c. SMZL demonstrated positive staining for CD1d
but negative staining for CD200 in 4/4 (100%) cases, and showed dim or negative
staining for CD25, CD103, and CD11c. Finally, marginal zone lymphoma involving
120
| EAHP - 2014
| 17-22 October 2014
[PP-LYMP-086]
DIFFUSE LARGE B CELL LYMPHOMA WITH AN UNUSUAL
IMMUNOPHENOTYPE: CD99 EXPRESSION MIMICS T-CELL
ORIGIN
Emoke Horvath, Brigitta Gnandt, Mihai Lazar Turcu
Department of Pathology, University of Medicine and Pharmacy Targu-Mures, Romania
Background: Diffuse large B-cell lymphoma (DLBCL) is the most common subtype
of NHL Intratumoral heterogeneity and variability of tumor cell’s immunophenotype
are important pitfalls in lymphoma diagnosis.Previously, we found CD99 expression
in a few extranodal DLBCL lymphoma cases, making it difficult to diagnose.
Methods: Tissue micro-arrays (TMA) were constructed for the paraffin blocks of
59 patients with de novo DLBCL with nodal presentation (all of them without well
defined subclass entities). TMA were tested by the immunoperoxidase method using
specific antibodies against CD20, CD10, Bcl-6, Ki 67 and CD99.
Results: CD99 expression with membrane staining was observed in 11 samples
(18,64%). Concommitant expression of CD99 and CD10/Bcl-6 antigenes was present in 9 cases (15,25%). Regarding the correlation of studied antigen with germinal center or non-germinal center derived immunophenotype, we have not found a
statistically significant correlation between CD99 expression and DLBCL subtypes.
All of CD99 positive DLBCL were associated with high Ki67 index (ranging between
60-90%).
Conclusions: LCA and CD99 positive lymphoid neoplasms with high Ki67 index do
not exclude the mature B-cell origin of the tumor, on the other hand, DLBCL should
be included in the differential diagnosis of CD99 positive nodal or extranodal tumors.
Keywords: DLBCL, TMA, CD99
PROGNOSTIC SIGNIFICANCE OF BONE MARROW
INVOLVEMENT PATTERN IN WALDENSTRÖM
MACROGLOBULINEMIA: ANALYSIS OF A SERIES OF 70
PATIENTS
PRIMARY LYMPHOMAS OF THE SPLEEN: 20 YEARS
EXPERIENCE IN A SINGLE CENTER
Alejandro Martín Martín1, Laura Magnano2, Hugo Álvarez Argüelles1,
Carolina De Bonis1, Taida Martín Santos1, Beatriz Soria1,
Sunil Lakhwani1, Eva Giné2, Sonia García Hernández1,
Miguel Teodoro Hernández García1, Luis Hernández Nieto1,
María Rozman2, José María Raya1
1
2
Hospital Universitario de Canarias. La Laguna, Tenerife, Spain
Hospital Clínic, Barcelona, Spain
Background and Aims: Waldenström macroglobulinemia (WM) is defined as a lymphoplasmacytic lymphoma with bone marrow involvement and an IgM monoclonal
gammopathy of any concentration. With a typically indolent course, factors associated with a worse prognosis include advanced age, cytopenias, functional status,
elevated beta-2 microglobulin level and, for some authors, a diffuse pattern of bone
marrow involvement. Our aim was to analyze the clinical and biological features of
patients with a WM diagnosis, with special attention to the clinical and prognostic
significance of bone marrow involvement pattern.
Methods: We studied 70 patients diagnosed between 1992 and 2013. Data collected at diagnosis were age, sex, presentation, hyperviscosity and B symptoms,
organomegaly, lymphadenopathy, blood count, peripheral expression, beta-2 microglobulin, serum IgM, serum LDH, lymphoplasmacytic infiltration in bone marrow aspirate, tumor burden and predominant marrow pattern (interstitial, nodular
or diffuse) in bone marrow trephine, and cytogenetics. We also collected treatment schemes and response, transformation to aggressive lymphoma, overall
survival (OS), and mortality. Statistical analysis was performed using SPSS and
Statxact programs.
Results: Mean age was 67 years (range 37-92), male 63%. The presence at diagnosis of lymphadenopathy, splenomegaly, hepatomegaly and peripheral expression
was 29%, 11%, 7% and 11%, respectively. Serum LDH and beta-2 microglobulin
were found elevated in 10% and 30% of cases each. The average values of tumor
burden in bone marrow biopsy and lymphoplasmacytic infiltration in aspirate were
43% (range 5-95) and 40% (5-96), respectively. Only two cases of transformation to
aggressive lymphoma were observed. The predominant pattern was interstitial in 31
cases (44%), nodular 23 (33%) and diffuse 16 (23%). In 11 of 23 cases with a nodular pattern (48%), paratrabecular aggregates were also present. A diffuse pattern
was correlated with lower platelet counts (p=0.005), lower values of hemoglobin
(p=0.003), and a higher frequency of B symptoms (p=0.047). The median OS was
68 months (range 1-247). The type of marrow involvement pattern did not influence
the OS and, specifically, the diffuse pattern was not associated with a worse prognosis. Mortality correlated with elevated serum LDH (p=0.046). We observed a strong
statistical correlation between older age and a worse OS (p<0.001); the presence
of B symptoms (p=0.046) and an elevated serum beta-2 microglobulin (p=0.037)
were also related with shorter OS.
Conclusions: In our experience, in line with other authors, an elevated serum beta2 microglobulin and, mainly, advanced age, are adverse prognostic factors in WM.
However, the pattern of bone marrow involvement does not influence the OS, and
specifically the diffuse pattern does not carry a worse prognosis when compared
with other patterns.
Keywords: Waldenström macroglobulinemia, bone marrow biopsy, prognosis
Iva Borisova, Alejandro Martín Martín, Carolina De Bonis,
Taida Martín Santos, María José Rodríguez Salazar,
Beatriz Soria, Hugo Álvarez Argüelles, Patricia Pecos,
Miguel Teodoro Hernández García, Luis Hernández Nieto,
José María Raya
Hospital Universitario de Canarias. La Laguna, Tenerife. Spain
Background: Primary lymphomas of the spleen constitute a rare subset of B-cell
lymphoproliferative disorders, being “classic” and “variant” hairy-cell leukaemia (cHCL and v-HCL) and splenic marginal zone lymphoma (SMZL) the most common.
An indolent course and certain clinical and biological heterogeneity (some degree of
overlap included) are significant features.
Patients and Methods: A total of 35 patients (18 SMZL, 15 c-HCL and 2 v-HCL)
were diagnosed in our center from 1994 to 2013. We retrospectively noted, at
diagnosis, age and sex, presentation, splenomegaly and lymphadenopathy, blood
count parameters, cell morphology, serum lactate dehydrogenase (LDH) and beta-2
microglobulin, viral serologies, direct Coombs test, and serum immunoglobulins.
We have studied the characteristics of bone marrow aspirate and trephine biopsy.
The presence of chromosomal abnormalities and cell immunophenotype by flow
cytometry or by immunohistochemistry were also collected. Splenomegaly was determined by both physical examination and imaging tests (ultrasound and computed
tomography), and the extra-splenic involvement by PET. The performance or not
of splenectomy and the first-line treatment approaches (including “wait and see”
attitude) were recorded. Finally, we also noted disease-free survival (DFS), overall
survival (OS) and mortality. The statistical analysis was performed using SPSS software updated for Windows.
[PP-LYMP-088]
[PP-LYMP-087]
Results: When compared with c-HCL, patients with SMZL were older (50 vs. 64year old; p=0.011) and presented more frequently with abdominal discomfort (7%
vs. 38%), elevated serum LDH (7 vs. 39%; p=0.046), a trend towards an elevated
serum beta-2 microglobulin (10% vs 41%; p=0.087) and a more frequent presence
of M-protein (0% vs. 22%; p=0.069). Leucopenia, neutropenia and thrombocytopenia were more frequent in patients with c-HCL (73%, 73% and 80%, vs. 28%, 18%
and 61%, respectively), while leukocytosis was predominant in SMZL (45 vs. 13%).
Main differences in phenotype were related to a net positive expression of CD103
and CD25 in c-HCL in comparison with SMZL (100% vs. 17%: p=0.001; and 100%
vs. 42%: p=0.005, respectively). Dry-tap marrow aspiration was observed in 58% of
c-HCL and never in SMZL (p=0.001), and an intrasinusoidal bone marrow infiltration
was more frequent in SMZL (33% vs. 0%: p=0.042). Splenectomy was perfomed
at diagnosis in 50% of SMZL cases and only in 8% of c-HCL (p=0.02). In HCL,
expectant attitude (38%) and cladribine (31%) were the most frequent front-line
approaches, while in SMZL R-CHOP scheme (39%) and expectant attitude (33%).
Despite a high degree of heterogeneity in the therapeutic approach over the years,
there were no differences in terms of DFS and OS and only one c-HCL and two
SMZL patients have died. The small number of patients with v-HCL did not allow
comparisons with the other two groups.
Conclusions: In reviewing our results we found some clinical and biological features
which may help to differentiate the two diseases, as well as a high degree of heterogeneity in the treatment of patients. The performance of splenectomy was much
more common in SMZL due to greater diagnostic difficulty and greater clinical need.
The observed survival in our series is very long, as already noted in the literature,
but despite the “indolent” course of these lymphoproliferative disorders, we think
that therapeutic criteria must be unified, for which, prospective multicenter clinical
trials are required.
Keywords: Hairy cell leukemia, splenic marginal zone lymphoma, splenic lymphoma
| 17-22 October 2014
| EAHP - 2014
|
121
th
[PP-LYMP-089]
MANTLE CELL LYMPHOMA WITH AN IN SITU MANTLE
COMPONENT: REPORT OF TWO CASES
Barbara Christoforidou1, Persefoni Xirou1, Sotirios Barbanis1,
Dimitrios Minotakis1, Pavlina Konstantinidou2, Frideriki Patakiouta1
(OS) of 90% and a 5 year disease free survival (DFS) of 81%. The clinical variables
(R-IPI, stage), the cellular subtype (germinative center B-cell or non-germian center
B-cell ) defined by Hans and Choi IH algorithms, high c-MYC and BCL2 protein
expressions (c-MYC>20% and BCL2>40%) and TLG did not show significant correlations with disease progression. High Ki67 index (cutoff:67%), high SUVmax (cutoff
value: 13,77) and low MTV values (cutoff value: 234 cm3) were significantly associated with longer DSF (p=0,002, p<0,0001 and p=0,002 respectively).
We report two cases of mantle cell lymphoma, which coexist with an in situ mantle
component.
Conclusion: Our results indicate that the MTV of pretreatment PET is a potential
prognostic marker for disease progression in R-CHOP treated DLBCL patients. The
significant correlation of high proliferation index and high SUVmax with longer DFS
are in contradiction with previous results published in the literature and might reflect the effectiveness of the chemo-immunotherapy of high grade tumors with
higher tumor cell growth fractions.
Our patients are an 85 and a 66 years old men.
Keywords: diffuse large B-cell lymphoma, PET/CT scan, prognostic factors
1
Department of Pathology Theagenion Cancer Hospital, Thessaloniki, Greece
Department of Haematology Theagenion Cancer Hospital, Thessaloniki, Greece
2
The first one presented with vomiting, anemia and weight loss. Clinical investigation
showed general lymphadenopathy. Biopsy of a right axillary lymph node revealed
an overt mantle cell lymphoma. Additionally an “ in situ mantle cell lymphoma” was
indicated in a gastric biopsy as an incidental, simultaneous finding.
Table 1. Univariate analysis using Kaplan-Meier curves
Prognostic marker, cutoff
The other patient presented with diffuse lymphadenopathy and fever. Biopsy of a
right inguinal lymph node revealed a mantle cell lymphoma with a mantle zone
growth pattern.
27
96
MTV>234 cm3
21
62
SUVmax<13,7
14
50
Cases diagnosed as in situ mantle cell lymphomas include two distinct groups with
different clinical implications.
SUVmax>13,7
34
94
TLG<16975
24
92
Most patients present with a localized enlarged lymph node or extranodal lymphoid
tissue. The normal architecture is preserved and cyclin-D1 positive cells are restricted to mantle zones of otherwise normal appearing follicles. The restricted distribution of the atypical cyclin-D1 positive cells to the mantle zone of follicles suggests
that these lesions may represent an early step in the development of mantle cell
lymphoma. However some patients, like in our first case, have evidence of more
widespread disease.
The second group, like our latter case, are overt mantle cell lymphomas with a
mantle zone growth pattern, that may have disseminated disease at diagnosis.
Keywords: In situ mantle cell lymphoma
TLG>16975
24
71
Ki67index<67%
17
65
Ki67index>67%
22
100
c-myc<20%+bcl2<40%
4
100
c-myc>20%+bcl2>40%
25
80
Total/overall
48
81
p-value
[PP-LYMP-091]
PROGNOSTIC BIOMARKERS FOR DISEASE PROGRESSION IN
R-CHOP TREATED DLBCL PATIENTS
EOSINOPHILIC FOLLICULITIS OCCURRING AFTER BONE
MARROW AUTOGRAFT IN A PATIENT WITH CLL
Botond Tímár1, Tamás Györke2, Dávid Molnár3, Judit Demeter4,
Lajos Gergely5, Tamás Masszi6
Erman Ozturk1, Ebru Zemheri2, Seyma Ozkanlı2,
Ayse Serap Karadag3, Tulay Zenginkinet2, Abdullah Aydın2
Semmelweis University, 1st Department of Pathology and Experimental Cancer Research,
Budapest, Hungary
2
Semmelweis University, Department of Nuclear Medicine, Budapest, Hungary
3
Semmelweis University, Faculty of Medicine, Budapest, Hungary
4
Semmelweis University, 1st Department of Internal Medicine, Budapest, Hungary
5
University of Debrecen, Institute of Internal Medicine, Division of Haematology, Debrecen, Hungary
6
St. István and St. László Hospital, Department of Haematology and Stem Cell Transplantation,
Budapest, Hungary
Aims: Prognostic factors for diffuse large B-cell lymphoma treated with standard
R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, prednisolone)
regimen were investigated in 48 newly diagnosed DLBCL patients (median age
55 years, range 18-81). The prognostic value of immunohistochemical (IHC) biomarkers, namely Ki67, c-MYC and BCL2 protein expressions and the following 3
quantitative parameters of pretreatment 18F-FDG PET/CT scans were evaluated:
maximum standardised uptake value (SUVmax), whole-body metabolic tumor volume (MTV) and total lesion glycolysis (TLG).
Materials-Methods: 48 patients with newly diagnosed DLBCL underwent PET before standard R-CHOP treatment. MTV and TLG were measured with gradient-maximizing region growing algorithm included in the Interview Fusion software package (Mediso Ltd, Hungary). Immunohistochemical stainings was performed on 5μm
sections using standard procedures. GCB and non-GCB phenotypes were defined
by Hans and Choi IH algorithms. Receiver operating characteristic (ROC) analysis
was performed to determine optimal cut-off values for MTV, TLG, SUVmax and Ki67.
The published cut off values were used for c-MYC and BCL2. Estimates of diseasefree survival (DFS) in the different patient groups were calculated according to the
Kaplan-Meier method and compared with the log-rank test.
| 17-22 October 2014
0,0001
0,08
0,002
0,702
1
Department of Hematology, Medeniyet University Göztepe Training and Research Hospital, İstanbul,
Turkey
2
Department of Pathology, Medeniyet University Göztepe Training and Research Hospital, İstanbul,
Turkey
3
Department of Dermatology, Medeniyet University Göztepe Training and Research Hospital,
İstanbul, Turkey
Eosinophilic pustular folliculitis (EPF) is a rare disorder. Recently, many EPF cases
have been observed in patients affected by acquired immunodeficiency syndrome
and EPF is rarely associated with other immunologic malfunctions such as lymphoma and leukemia. Generally, the prognosis for EPF is good when the EPF occurs after auto-bone marrow transplantation (BMT) and is treated with oral corticosteroids.
We report a patient who was diagnosed as chronic lymphocytic leukemia and eosinophilic folliculitis. The skin lesions were seen 4 months after allo-BMT while under cyclosporine medication. Skin biopsy specimen revealed numerous eosinophil
infiltrations from the hair follicles to the sebaceous glands. No sign of skin graftversus-host disease (GVHD) was found.
EPF after BMT is a distinctive entity and it may be an extremely rare occurrence
and it is clinically indistinguishable from ordinary seborrheaic eczema. As reported
previously, it is important that this skin lesion must be distinguished from the signs
of skin GVHD.
Keywords: Eosinophilic pustular folliculitis (EPF), bone marrow transplantation
(BMT), CLL
Results: After a median follow-up time of 43 months (range 6-71 months) 5/48
patients died and 9/48 patients developed relapse resulting a 5 year overall survival
| EAHP - 2014
0,002
Abreviations: DFS: disease free survival; MTV: metabolic tumor volume; SUVmax: maximum
standardised uptake value; TLG: total lesion glycolysis.
[PP-LYMP-090]
1
122
Number of cases 5 year DFS %
MTV< 234 cm3
Fina Climent1, Gloria Pérez Esteve4, Ramon Bosch2, Mar Varela1,
Cristina Muniesa3, Enric Condom1, Octavi Servitje3
1
Department of Pathology, Hospital Universitari de Bellvitge-IDIBELL, L’Hospitalet de Llobregat
(Barcelona), Spain
Department of Pathology, Hospital Verge de la Cinta-IISPV-FF, Tortosa (Tarragona), Spain
3
Department of Dermatology, Hospital Universitari de Bellvitge-IDIBELL, L’Hospitalet de Llobregat
(Barcelona), Spain
4
Department of Pathology, Hospital Universitari de Bellvitge-IDIBELL, L’Hospitalet de Llobregat
(Barcelona), Spain; Department of Pathology, Hospital Universitari de Girona Dr. Trueta, Girona, Spain
2
Objective: To study the clinicopathological, immunophenotipical features and molecular findings of B-cell cutaneous lymphoid hyperplasia (CLH) triggered by the
administration of vaccines or allergens adsorbed by aluminium.
Methods: The clinical, pathological, immunophenotipical and molecular findings of
vaccine-induced B-cell CLH cases were reviewed.
Results: Our series comprised 5 women with a median age of 40 years old (range:
25-56). Three of them had had tetanus vaccinations and two had had specific immune therapy. The delay between vaccination and the onset of skin symptoms was
variable, ranging from 1 to 8 years. 2 patients had disseminated lesions.
Primary and disseminated lesions showed deep dermal and hypodermal lymphoid
follicular infiltrates, with germinal center formation that displayed immunophenotipic features of reactive germinal centers, mostly composed of B cells without atypia
(CD20+). The germinal centers were positive for BCL6 and CD10 and negative for
BCL2. The rare plasma cells showed a polyclonal expression of kappa and lambda
light chains. Studies of the clonality status revealed polyclonal patterns of IgH gene
rearrangement.
Conclusion: B-cell CLH is a rare complication of vaccination that clinically and histologically mimics cutaneous B-cell lymphoma. Primary and disseminated lesions did
not show any histological, immunophenotipical and molecular differences.
Keywords: B-cell pseudolymphoma, vaccination
Conclusions: Our findings from this case report demonstrate for first time that mutations
of the 266R codon of p53 gene may be an early (driver) gene mutation involved in FL
transformation. Therefore, this case along with previously reported data for the potential
predictive value of p53 mutations in FL at diagnosis, support the use of immunohistochemical and molecular testing of p53 gene in all newly diagnosed low grade FL.
Keywords: Follicular lymphoma, p53, transformation
[PP-LYMP-094]
GENE EXPRESSION PROFILING IN HODGKIN’S LYMPHOMA CELL
LINES AND EPSTEIN-BARR VIRUS (EBV)-POSITIVE VERSUS
EBV-NEGATIVE CLASSICAL HODGKIN’S LYMPHOMA SAMPLES
B-CELL CUTANEOUS LYMPHOID HYPERPLASIA (B-CELL
PSEUDOLYMPHOMA) RELATED TO VACCINATION: STUDY OF
FIVE CASES
Results: A 66-year old patient with a previous history of rheumatoid arthritis treated
with rituximab and a TNF-alfa-blocking agent, presented with B-symptoms and a
large tumor mass in abdomen with infiltration of the omentum. At presentation, LDH
was 3,8 microkat/L and s-albumin 34 g/L. Bone marrow was negative. A diagnosis
of stage III low grade follicular lymphoma was made with tumor cell proliferation
<5%. The tumor cells were positive for t(14;18) assessed by FISH. The patient initially received rituximab (4x) without response and subsequently CHOP-21 x 6 with
stable disease for 3 months. Because of clinical progression, the patient thereafter
received bendamustin followed by local radiation therapy. Thirty five months after
initial diagnosis, a new biopsy from abdomen confirmed transformation to DLBCL
with high tumor cell proliferation (90%). In both, diagnostic low grade FL and DLBCL
(transformed FL), a rare c.796G>A pG266R mutation of the p53 gene (exon 8) was
detected, which is reported for first time in FL. This mutation is rarely detected
in other lymphoma types accounting for <0.7% of all lymphomas (IARC database,
R17). In addition, a silent mutation in exon 4 of the p53 gene was found in both
samples. No mutations in EBF1 or MYD88 genes were detected in the lymph nodes
at diagnosis or at transformation of FL. Moreover, aberrant loss of CD45 (LCA) in
FL cells was seen in the diagnostic lymph node and DLBCL cells. By immunohistochemistry, overexpression of p53 protein (100% positive tumor cells) was detected
in both samples. No change in the levels of c-myc protein was seen.
[PP-LYMP-092]
Paloma Martin1, M. Jose Coronado2, Elvira Ramil3, Yolanda Vicente5,
Isabel Millan4, Carmen Bellas5
1
[PP-LYMP-093]
RAPID TRANSFORMATION OF A GRADE 1-FOLLICULAR
LYMPHOMA ASSOCIATED WITH AN UNCOMMON C.796G>A
PG266R MUTATION OF THE P53 GENE AT DIAGNOSIS BUT NOT
WITH MYD88 OR EBF1 GENE MUTATIONS
Anna Kwiecinska1, Mehran Ghaderi1, Ioanna Xagoraris1,
Andras Matolcsy2, Leonie Saft1, Anders Österborg3,
George Z. Rassidakis1
1
Department of Pathology and Cytology, Karolinska University Hospital & Karolinska Institute,
Stockholm, Sweden
First Department of Pathology and Experimental Cancer Research, Semmelweis University, Budapest,
Hungary
3
Department of Hematology, Karolinska University Hospital & Karolinska Institute, Stockholm, Sweden
2
Background: Transformation of low-grade follicular lymphoma (FL) to aggressive diffuse large B-cell lymphoma (DLBCL) has been associated with acquiring new genetic
events including p53 mutations during the disease course. In addition, the presence of
p53 mutation at the time of diagnosis of follicular lymphoma may identify a high-risk
group of patients with rapid disease progression and poorer survival (Blood 2008;
112:3126). Recent evidence suggests that additional mutations in several genes such
as EBF1 and MYD88 are gained at transformation (Nat Genet 2014; 46(2):176).
Purpose: We report a case of rapid follicular lymphoma transformation associated
with an uncommon p53 mutation, and loss of CD45 expression at diagnosis.
Methods: Histologic examination, immunohistochemistry, flow cytometry, molecular
diagnostics including PCR for IgH and light chain gene rearrangements, and fluorescent in situ hybridization (FISH) for t(14;18) translocation were used as diagnostic
methods for this case. Mutation analysis for p53 (exons 4-8), EBF1 and MYD88
genes was performed using standard PCR and automated direct sequencing techniques. Additional immunohistochemical studies were performed to assess p53 and
MYC protein levels.
Molecular Pathology Laboratory, Instituto de Investigación Sanitaria Hospital Puerta de Hierro, Madrid,
Spain
2
Confocal Microscopy Unit, Instituto de Investigación Sanitaria Hospital Puerta de Hierro, Madrid, Spain
3
Sequencing and Molecular Biology Unit, Instituto de Investigación Sanitaria Hospital Puerta de Hierro,
Madrid, Spain
4
Clinical Biostatistics Unit, Hospital Puerta de Hierro, Madrid, Spain
5
Department of Pathology, Hospital Puerta de Hierro, Madrid, Spain
There is long-standing recognition of a relation between Hodgkin Lymphoma (HL)
and Epstein-Barr virus (EBV), although the mechanisms underlying the contribution
of virus infection to the development and maintenance of the transformed phenotype remain to be established. It is known that EBV-encoded proteins mimic key
signalling pathways in B cells and seem to play a role in the development of the
lymphoma. LMP2A, an EBV encoded protein, is thought to contribute to the pathogenesis of HL. Two cHL cell lines (L428 and L1236) were infected with LMP2A using
lentiviral vectors and an initial oligonucleotide microarray was performed to compare the gene expression pattern between infected and non-infected cell lines. Eight
genes (GDF15, ATF5, GRK4, CENPE, MIR17HG, KIF18A, LAMB3, and DEK) differentially expressed were selected and further investigated for validation. Expression
levels of GDF15, ATF5, GRK4, CENPE, MIR17HG, KIF18A, LAMB3, and DEK were
analyzed in 24 cases of cHL; 12 EBV positive and 12 EBV negative, using RealTime
ready Custom Panels. The target gene expression was normalised by three non-regulated reference genes (HMBS, YWAZ, and SDHA) for relative analysis quantification.
Expression analysis showed high levels of ATF5 and DEK in most cHL cases. GDF15,
GRK4, CENPE, MIR17HG, KIF18A, and LAMB3 were down regulated. Many studies
have reported that DEK is implicated in several signalling pathways in tumour cells
and played an important role in cancer progression. ATF5 is a transcription factor of
the ATF/CREB protein family, and previous studies suggested that ATF5 is required
for the survival of cancer cells. DEK and ATF5 are up-regulated in our series of cHL;
its implication in cHL etiopathogeny needs further investigation.
When we compared the relative expression ratios, no significant differences were
found between EBV positive and EBV negative cases. Our results suggest that expression of the analyzed genes is not due to EBV infection; there is discordance
between cell lines results and human samples, probably owing to its histological
complexity, with a minor population of the neoplastic Hodgkin and Reed-Sternberg
cells diluted in a reactive inflammatory background.
Keywords: Hodgkin Lymphoma, Epstein-Barr virus, Gene expression
| 17-22 October 2014
| EAHP - 2014
|
123
th
[PP-LYMP-095]
NUCLEAR OPTICAL DENSITY IS HIGHER IN FOLLICULAR
LYMPHOMA GRADE 1 THAN IN MANTLE CELL LYMPHOMA AND
SMALL LYMPHOCYTIC LYMPHOMA
Dragan Slavoljub Mihailovic, Zaklina Zarko Mijovic
University of Nis, Faculty of Medicine, Centre of Pathology, Serbia
Introduction: Mantle cell lymphoma (MCL) is defined as a B-cell neoplasm composed of monomorphic small to medium-sized lymphoid cells with irregular nuclei
that morphologically resemble centrocytes but often have slightly less irregular
nuclear contours. Immunohistochemistry have allowed a more precise definition of
lymphomas and distinction from other types of of small B-cell lymphomas.
to adult women (50-65 years). Clonal IgH gene rearrangement by endpoint PCR was
positive in all cases. One case (1.7%) showed BCL2/IGH t(14;18) (Kreatech) by FISH
and was BCL-2 positive using E/17, being considered as pseudo negative BCL-2
(100/D) FL. The clinical course was favorable in all patients and all of them are currently asymptomatic. (Table 1 and 2)
Conclusions: BCL-2 negative follicular lymphomas are rare entities, however their
recognition has increased. In this work we demonstrate the utility of BCL-2 antibody
clone E17, to identify BCL2 pseudo negative FL with t(14;18). This study shows the
importance of new technologies to identify this particular subtype of FL (BCL-2 negative real and BCL-2 pseudo negative) and their utility in the differential diagnosis
between follicular hyperplasia and FL (BCL2 100/D negative), with the subsequent
clinical impact that this represents.
Keywords: Follicular lymphoma, BCL2 (100/D) negative, BCL2 (E17)
Table 1. Demographic and clinical characteristics
Aim: To estimate karyometric variables in nodal non-Hodgkin lymphomas composed of small B-cells.
Case Age Gender Clinical characteristics
Material and Methods: At Centre of Pathology, Clinical Centre of Nis, 5 cases of
mantle cell lymphoma, 7 cases of follicular lymphoma grade 1, and 10 cases of
small lymphocytic lymphoma (SLL) were diagnosed using appropriate histological
and immunohistochemical criteria. Using ImageJ software, on digital images obtained at x40 objective, seven nuclear variables were estimated: nuclear area, mean
optical density (OD), modal optical density, perimeter, Feret diameter, integrated optical density (IOD) and roundness.
Results. Nuclear area, perimeter and Feret diameter were significantly higher in mantle cell lymphoma than in SLL and follicular lymphoma grade 1. Modal nuclear optical
density was significantly higher in follicular lymphoma grade 1 than in mantle cell
lymphoma and SLL. Differences in nuclear shape were not statistically significant.
Conclusion: Karyometric analysis may provide additional information in differential
diagnosis of small B-cell lymphomas.
Treatment and clinical
course
1
63
Female
Cervical, axillar
and retroperitoneal
lymphadenopathy, as well as
B symptoms. Stage: IIIBx.
FLIPI: High risk.
CHOP.
Alive 14 months.
2
50
Female
Inguinal bulky, swelling
and pain. Stage: IIA. FLIPI:
Low risk.
CHOP.
Alive 14 months.
3
65
Female
Cervical lymphadenopathy.
Stage: IIA. FLIPI: Low risk.
RCHOP. Alive 15 months.
4
60
Female
Painful palpable inguinal
mass.
Without treatment, Alive 6
months.
FLIPI: Follicular lymphoma international prognostic index. R: Rituximab. CHOP: Cyclophosphamide,
doxorubicin, vincristine and prednisone.
Keywords: lymphoma, karyometry, density
Table 2. Immunohistochemistry and molecular profile
Case 1
Case 2
Case 3
Case 4
CD20
+
+
+
+
CD3
-
-
-
-
CD10
+
+
+
+
[PP-LYMP-096]
BCL-2 (100/D) NEGATIVE FOLLICULAR LYMPHOMA:
PATHOLOGICAL, IMMUNOHISTOCHEMICAL AND MOLECULAR
FEATURES IN 4 MEXICAN ADULT PATIENTS
Carmen Lome Maldonado1, Lorena Viramontes Aguilar1,
Jose Tovar Bobadilla1, Jorge Garcia Vera1,
Roberto Hernández Mora1, Braulio Martinez Benitez1,
Arturo Angeles Angeles1, Yvette Neme2, Leticia Quintanilla Fend3
Pathology department Instituto Nacional de Ciencias Medicas y Nutricion “Salvador Zubiran”
Mexico city, Mexico
2
Hematology department ABC Medical Center Mexico City, Mexico
3
Institute of Pathology, University of Tüebingen. Tüebingen Germany
Purpose: To determine the frequency of BCL-2 negative follicular lymphomas in
Mexican adult patients and to describe the clinical, prognostic and genetic profiles (using PCR, FISH and sequencing) in BCL-2 negative cases without t(14;18).
Furthermore, to assess the utility of the antibody BCL-2/E17 in BCL-2 negative
cases with or without t(14;18)
Materials-Methods: We reviewed 56 cases of FL in a 10 year interval. Four of these
were diagnosed as LF BCL-2 negative. We analyzed the clinical, histological and
immunophenotypical features using 2 antibody clones of BCL-2 (E/17 and 100 / D)
assessed by molecular (FISH, PCR, MBR/mcr) and sequencing tests for the presence
of BCL2/IGH t(14; 18) fusion, BCL-6/IGH t(3;14) (q27q32), and del(1p36).
Results: A total of 56 cases of FL were identified. Four cases (5.3%) were diagnosed
as FL BCL-2 negative (100/D clone, Biocare). All were FL grade 3A and corresponded
124
| EAHP - 2014
| 17-22 October 2014
-
-
-
-
BCL-2 (E/17 clone)
+
-
-
-
BLC-6
1
Background: Follicular lymphoma (FL) has a germinal center origin, it is the second
Non-Hodgkin´s lymphoma in frequency and it is classified into 3 histological grades,
based on centroblasts percentage and the presence or absence of centrocytes. The
t(14;18) (q32, q21) is reported in 85% of cases and it is associated with overexpression of BCL-2. Nevertheless, there are BCL-2 negative FL subtypes with t(14;18);
this cases are considered as “pseudo negative BCL-2 (100/D) follicular lymphomas”
and exhibit different acquired somatic mutations. Another subtype of LF is BCL-2
negative, which lacks t(14;18). This subgroup is considered as BCL2 real negative
FL and its genetic and molecular characteristics are different from the classic form
that has BCL-6 translocations and del(1p36).
BCL-2 (100/D clone)
+
+
+
+
Ki67
10-50%
>75%
30%
70%
FISH
t(14;18) (q32;q21)
*
Negative
Negative
Present
Present
Present
Present
**
*
*
*
Clonal IgH gene
rearrangement
(PCR)
Secuencing
*In course. **Not necessary.
[PP-LYMP-097]
C-MYC AND BCL-2 TRANSLOCATION FREQUENCY IN DIFFUSE
LARGE B CELL LYMPHOMAS
Bahar Akkaya1, Ozan Salim2, Hampar Akkaya3, Mualla Ozcan1,
Orhan Kemal Yucel2, Ramazan Erdem2, Utku İltar2, Levent Undar2
1
2
3
Akdeniz University School of Medicine Pathology Department, Turkey
Akdeniz University School of Medicine Hematology Department, Turkey
Bașkent University School of Medicine Pathology Department, Turkey
Introduction: Diffuse large B-cell lymphoma (DLBCL) is the most common type of
malignant lymphoma and constitutes 30-40% of adult non-Hodgkin’s lymphomas in
western countries. Diffuse large B-cell lymphomas (DLBCLs) are a biologically heterogeneous group with varied clinical courses, histology, molecular and cytogenetic
characteristics. Recent studies showed that DLBCL cases with gene expression profiles similar to germinal center B cells had much better prognosis than DLBCL cases
with gene expression profiles resembling activated B cells. 3 antibody panel (CD 10,
Materials-Methods: In the present study, we evaluated the expression patterns of
CD 10, BCL6 and MUM 1 by immunohistochemistry on paraffin-embedded tissues
from 70 DLBCL patients. MYC, BCL2 rearrangements were investigated by interphase fluorescence in situ hybridization on tissue microarrays in 47 DLBCLs.
Results: Mean age of patients is 56.1 year. There were 34 women (48,5%) and 36
men (51,5%) patients. Tumors were located at lymph nodes in 30 patients (42,85%).
There were 34 GCB subtypes and 36 ABC sub types DLBCL cases.
C-MYC translocation was observed in 6,3 % (3/47). (2; GCB, 1; ABC) Two of them
were extranodal lymphoma, one case was nodal lymphoma. No relationship between
C-MYC translocation and/or age, gender, DLBCL subtype, stage was observed.
BCL2, rearrangements were detected in (6/47) DLBCL patients. Five of them were
extranodal lymphoma, one case was nodal lymphoma. No relationship between
C-MYC translocation and/or age, gender, DLBCL subtype, stage was observed.
One case showed a combination of c-MYC and Bcl-2 rearrangements in 47 DLBCL
cases. The patient was male, 63 years old. The case was GCB type of DLBCL. The
disease recurred in 19. month and the patient was died in 35. month after diagnosis.
Conclusion: Cases in which clinical, morphological and immunohistochemical diagnosis are not adequate to discriminate DLBCL-BL, cytogenetic studies are beneficial. In this way, not only the patient will be able to reach the correct diagnosis but
also get the effective treatment. We concluded that C-MYC may contribute to aggressive transformation, and more mechanism-based therapy should be explored.
Keywords: C-MYC translocation, Bcl-2 translocation, lymphoma
[PP-LYMP-098]
THE PREVALENCE OF EPSTEIN-BARR VIRUS INFECTION IN
FOLLICULAR LYMPHOMA IN KOREA: SINGLE CENTER 22 YEAR
EXPERIENCES
CUTANEOUS INTRAVASCULAR NK/T-CELL LYMPHOMA MIMICKING
ERYTHEMA NODOSUM AND DEEP VEIN THROMBOSIS - A CASE REPORT
Masaru Hosone1, Satoru Arai1, Naoyuki Higashi2, Shin-ichi Osada2,
Zenya Naito1
1
2
Nippon Medical School, Department of Pathology, Tokyo, Japan
Nippon Medical School, Department of Dermatology, Tokyo, Japan
Introduction: Intravascular lymphoma is a rare neoplasm and well known for a cause
of fever of unknown origin. Almost all cases of IVL are of B-cell origin and introduced
in WHO classification as intravascular large B-cell lymphoma. We herein report an extremely rare case of NK/T-cell IVL showing deep vein thrombosis (DVT)-like symptoms.
Clinical Course: A 66-year-old Japanese female visited our hospital with erythema and
painful subcutaneous nodules on left upper arm and left thigh and erythema nodosum
and vasculitis were suspected. Under a close follow-up at the outpatient skin clinic,
the right thigh was also swelled suddenly and severe gait disturbance was triggered.
The patient was admitted urgently and DVT, adult Still disease, collagen diseases and
related malignancies including hematolymphoid neoplasms were also suspected.
Pathologic Findings and Diagnosis: The left thigh skin biopsy revealed some small vessels in deep dermis and subcutis were filled with medium/large-sized atypical lymphoid
cells and fibrinoid necrotic tissue. These lymphoid cells showed positivity for CD2, cCD3,
CD30, CD43, CD45, CD56, Granzyme B, TIA-1, EBER1-in-situ hybridization and negativity
for CD5, CD7, CD10, CD20, CD23, CD79a, PAX5. The combined findings of the morphology and immunostaining reached the diagnosis of IVL with NK/T-cell immunophenotype.
Discussion: Our case should be included in extranodal NK/T-cell lymphoma, nasal
type in recent WHO classification and also categorized as an unusual type of IVL.
The common IVL with B-cell origin itself is a rare form of mature B-cell lymphoma
and IVL with NK/T-cell origin is an extremely rare disease and only a limited number
of case reports have been published. They reported female predoninance, earlier
onset, common cutaneous manifestation with EBV infection and poor prognosis. Our
case showed typical features except for an old age and survival after multi-drug
chemotherapy. We report this time a distinctly rare and interesting case of NK/T-cell
IVL with a comprehensive review of reported cases.
The aim of this study was to investigate the frequency and prognostic impact of
BCL2 and MYC rearrangements.
[PP-LYMP-099]
BCL6, MUM 1) of immunohistochemical stains can be used to subclassify DLBCL into
prognostically significant germinal center B-cell like (GCB) DLBCL and activated B-cell
like (ABC) DLBCL subtypes. Both CD 10 and BCL6 are considered as germinal center
markers while MUM 1 is expressed in activated B-cells and plasma cells.
Keywords: Intravascular lymphoma, NK/T-cell lymphoma, skin
Eun Mi Son, Jooryung Huh, Heounjeong Go
[PP-LYMP-100]
University of Ulsan College of Medicine, Asan Medical Center, Seoul, South Korea
Background: Follicular lymphoma(FL) is most common type of lymphoma. The frequency is relatively lower in East Asia. However recently in korea the relative frequency of FL is increasing in past decade. But the clinicopathologic chatacteristics
of FL in korea has not been well studied. Epstein-Barr virus is an important transforming virus for B cells and associated with several lymphoproliferative disorders.
But study has not been conducted up till now to establish the prevalence of EBV
infection in patients of FL in Korean population. The aim of this study was to investigate the characteristics of FL in korea with regard to prevalence of EBV infection.
Materials and methods: 159 cases with the diagnosis of follicular lymphoma form
January 1991 to March 2014 in Asan Medical center. The histologic sections of all cases are reviewed by three pathologists and medical records were reviewed for clinical
characteristics. Using automated in situ hybridization (ISH) instrument( Benchmark,
ventana Medical systems) EBV-encoded small RNA was examined in 123 cases.
Results: FL in korea is more common in men (57.9%) than women (42.1%) with
younger median age about 49 years. Disease staging(Ann Arbor stage) in 158 patients included 41 patients (25,9%) with stage I disease, 13 (8.2%) with stage II,
33(20.9%) with stage III, and 71(44.9%) with stage IV. The cases were classfied as
grades 1 (63 patients, 39.6%), 2 (25 patients 15.7%), 3A (47 patients, 29.6%), and
3B ( 24 patients, 15.1%). The 5-year survival rate was 81.0%.In 152 of 159 cases
the FLIPI index was assessed which are distributed as follows; low risk (70 patients,
46.1%), intermediate risk ( 43 patients, 28.3%), poor risk ( 39 patients, 25.6%).
According to the FLIPI grade, the patients with low and intermediate risk group had
a better survival rate than patients with poor risk group ( log-rank test, p<0.001).
However, WHO grade was not statistically significant with regard to survival. EBVencoded small RNA are positive in 67(54.5%) of 123 cases.
HISTIOCYTIC SARCOMA OF STOMACH: CASE REPORT
Aydan Kilicarslan1, Ozge Basaran Aydogdu1, Hayriye Tatli Dogan1,
Gulnur Guler2
1
Department of Pathology, Medical School, Atatürk Training and Research Hospital, Yıldırım Beyazıt
University, Ankara, Turkey
2
Department of Pathology, Medical School, Hacettepe University, Ankara, Turkey
Background: Histiocytic sarcoma (HS) is a rare neoplasm of the lymphohematopoietic
system. HS presents with histiocytic differentiation both morphologically and immunophenotypically. HS is preferentially seen gastrointestinal tract, skin and other extranodal sites.
Case: 52 years old male patient presented with dyspepsia and weight loss. Upper GI endoscopic examination showed a giant ulcer lesion comprise fundus and proximal corpus
of stomach. Radiologically there was a hypodense tumor mass on the stomach wall with
34 mm thickness which was also present in perigastric fatty tissue by computerized tomography. A diagnosis of poorly differentiated neoplasm was made on endoscopic biopsy
of lesion. Patient underwent gastrectomy operation. Resection specimen examination
revealed an ulcerative tumoral lesion located on corpus and fundus. Grossly tumor infiltration was seen in subserosal fat. Microscopically non-cohesive, sheet-like intiltration of
atypical round/oval cells with abundant eosinophilic cytoplasm. Neoplastic cells also had
irregular nuclear contours with prominent nucleoli. Signifigant proportion of the cells were
bi-nuclear and some of them in giant morphology. Immunohistochemically diffuse CD68
and focal CD45 and lysozyme positivity were seen in neoplastic cells. Cytokeratins, S100
and myeloperoxidase were negative. After histiocytic sarcoma diagnosis chemotherapy
protocol was given to the patient. Patient died eighteen month later from diagnosis.
Conclusion: This is the largest series of Korean FLs. We found clinicopathologic
features of FL in korea was similar to the global trends with some differences. High
grade FL (,71/159, 44.7%) is common while bone marrow involvement is uncommon. The frequency of EBV associated FL is common.
Conclusion: HS cases located on stomach, colon, ileum and anal region were reported in literature. Systemic spread potential is quite high. HS has poor prognosis
on lagre proportion of the cases. There is also limited response to the classic treatment protocols. Diagnosis of HS can also be challenging and immunohistochemical
studies may help in suspected cases.
Keywords: Follicular lymphoma, Epstein-Barr virus
Keywords: Histiocytic sarcoma, stomach
| 17-22 October 2014
| EAHP - 2014
|
125
th
[PP-LYMP-101]
[PP-LYMP-103]
MORPHOLOGICAL PATTERN OF NODAL MARGINAL ZONE
LYMPHOMA IN EGYPT
SIGNET-RING-CELL VARIANT OF FOLLICULAR LYMPHOMA:
A CASE REPORT
Howayda Sayed Abd El All1, Dalia Ahmed Nafeeh2,
Rasha Abdelmotagally3
Tatjana Terzic1, Jelena Petkovic2, Snezana Sretenovic3,
Novica Boricic1, Vladimir Jurisic4
1
Pathology Department, Faculty of Medicine, Suez Canal University, Ismaliya; and
Immunohistochemistry Laboratory, Nasser Institute, MOH, Cairo, Egpyt
2
Hemato-Oncology Department, Faculty of Medicine, Alexandria University, Alexandria, Egypt
3
Oncology Department, Nasser Institute, MOH, Cairo, Egpyt
1
The aim of this study is to evaluate the morphological pattern of nodal marginal zone
lymphoma. Seventy five cases were included based on the WHO criteria (2008), 31
males and 26 females, 19-77 years, average 48 years.
Background: Signet-ring-cell (SRC) lymphoma is a rare variant of mature B and
T-cell lymphomas. We present a case of SRC follicular lymphoma, which highlights
diagnostic difficulties.
The following were studied: 1- Four architectural patterns, diffuse, nodular, parafollicular and interfollicular; 2- monotonous vs pleomorphic; 3- presence of monocytoid cells; 4- number of large cells estimated as <10%, 10-20%, > 20%; 5- number
of Ki67+ CD20+ cells estimated as <10%, 10-20%, > 20%; 6- pattern of follicular
dendritic cells (FDC) stained by CD35 and CD23 and evaluated as absent, shrunken
and expanded irregular.
Case: A 50-year-old female initially presented with isolated enlargement of inguinal
lymph node. Excisional biopsy was performed. Morphologic analysis revealed effacement of lymph node architecture by neoplastic follicles. The neoplastic cells were predominantly small, cleaved centrocytes, with scattered centroblasts (0-15 centroblasts/
hpf). Many of small cells showed cytoplasmic vacuolization. These clear vacuoles were
PAS and Alcian blue negative. The diagnosis was obtained on review of hematoxylin
and eosin stained paraffin-embedded slides (Fig.1) and immunohistochemical data
(Cytokeratin, S-100, Pax-5, CD20, CD3, CD5, CD23, CD43, CD10, bcl-2, bcl-6, CD138,
cyclin D1, IgG, IgM, kappa, lambda, Ki-67). These SRCs have intrafollicular distribution
and were positive for Pax-5 (Fig.2), CD20, CD10, bcl-2 (Fig.3), bcl-6 and IgG. Follicular
lymphoma, grade 1-2 (SCR variant) was diagnosed. After diagnosis the patient was
treated by radiotherapy, with clinical resolution of the affected inguinal node. But, one
year later, paraaortal lymph nodes were enlarged and second biopsy was performed.
The morphological and immunohistochemical finding was similar to the previous biopsy. The patient received combination immunochemotherapy (8 cycles of R-CHOP)
and complete remission was achieved.
None of the studded features correlated with either the age or the sex. The diffuse
pattern (28 cases) showed cellular pleomorphism, and absent FDC (p<0.005). The
perifollicular pattern (13 cases) was also pleomorphic, associated with expanded irregular FDC, increased number of large cells and ki67+ cells (p<0.005). The nodular
pattern (12 cases), was monomorphic, formed of small to medium size irregular
cells and shrunken FDC (p<0.005). The interfollicular pattern (4 cases), was pleomorphic and lacks any association with the other factors. The monocytoid cells
didn’t correlate with any pattern.
The association of these patterns with patients survival/ disease progression and
role of viral infection especially hepatitis B/C is currently under evaluation and will
be discussed.
Institute of Pathology, Faculty of Medicine, University of Belgrade, Belgrade, Serbia
Opsta bolnica Cuprija, Serbia
Clinic for hematology, Clinical Center Kragujevac, Serbia
4
Faculty of Medical Sciences, University of Kragujevac, Serbia
2
3
Keywords: architectural pattern, FDC, Ki67
Conclusion: SRC morphology is the most frequently reported in follicular lymphoma,
but also in extranodal marginal zone lymphoma, small lymphocytic lymphoma, diffuse large B-cell lymphoma and T-cell lymphoma. It has been described in nodal,
but also in a variety of extranodal locations. Therefore, it is important to avoid erroneous diagnosis of metastatic SRC carcinoma.
[PP-LYMP-102]
Keywords: Signet-ring, lymphoma
DETECTION OF EPSTEIN-BARR VIRUS (EBV) IN GASTRIC
DIFFUSE LARGE B CELL LYMPHOMA CASES
Aydan Kilicarslan1, Mehmet Dogan2, Hayriye Tatli Dogan1,
Nuran Sungu1, Hayriye Ergin Akkoz2
1
Yıldırım Beyazıt University Ankara Atatürk Education and Research Hospital, Department of
Pathology, Ankara, Turkey
2
Dr. Abdurrahman Yurtaslan Ankara Oncology Education and Research Hospital, Department of
Background: There are numerous studies about the role of Epstein - Barr virus in
the pathogenesis of lymphomas. In this study we evaluated the EBV status of gastric
diffuse large B cell lymphomas (DLBCL) from two institutions.
Figure 1. bcl-2x400.
Materials-Methods: Gastric DLBCL cases retrieved from the databases of Yıldırım
Beyazıt University Ankara Atatürk Education and Research Hospital, Department of
Pathology and Abdurrahman Yurtaslan Ankara Oncology Education and Research
Hospital, Department of Pathology between years 2008 and 2013. 23 gastric
DLBCL cases reviewed according to WHO 2008 classification. The presence of
Epstein–Barr virus small ribonucleic acids was examined by in situ hybridization using Epstein-Barr virus (EBV)-encoded small RNA (EBER) oligonucleotides
in automated platform.
Results: There are 10 male and 13 female patients. Mean age was 60.6. There were
two positive cases (8.7%) by EBER in situ hybridization.
Figure 2. HEx400.
Conclusion: EBV positive DLBCL is not common in Turkish population. Percentage of
EBV positivity was 8.6% and 5.3% in two previous studies. Although we had small
number of cases in our study, we found concordant result with previous Turkish
reports. Further studies are needed to evaluate the importance of EBV positivity in
gastric DLBCL in terms of etiology, prognosis and treatment.
Keywords: DLBCL, EBER, Gastric
Figure 3. pax5x1000.
126
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WITHDRAWN
Keywords: follicular lymphoma, CD5, t(14;18)
[PP-LYMP-105]
B-CELL LYMPHOMA, UNCLASSIFIABLE, WITH FEATURES
INTERMEDIATE BETWEEN DIFFUSE LARGE B-CELL LYMPHOMA
AND CLASSICAL HODGKIN LYMPHOMA WITHOUT MEDIASTINAL
DISEASE: 4 CASES MIMICKING NODULAR SCLEROSIS
CLASSICAL HODGKIN LYMPHOMA
Nurhan Sahin1, Ibrahim Sari2, Zehra Bozdag2,
Ozge Dilara Colakkadioglu2
1
2
İnönü University, Malatya, Turkey
Gaziantep University, Gaziantep, Turkey
B-cell lymphoma, unclassifiable, with features intermediate between diffuse large
B cell lymphoma and classical Hodgkin lymphoma (BCLu-DLBCL/CHL) is a diagnostic provisional category in the World Health Organization (WHO) 2008 classification
of lymphomas. It is also known as gray-zone lymphoma, has overlapping clinical
and biological characteristics of both diffuse large B-cell lymphoma and classical
Hodgkin lymphoma (CHL). These lymphomas are most commonly associated with
mediastinal disease. Similar cases are less commonly in the peripheral lymph node
groups as a primary site. The primary extranodal involvement is rarely seen. BCLuDLBCL/CHL is described by the WHO into four patterns along with detailed clinical,
morphological and immunophenotypic characterization and outcome data. In this
report, we present four cases with BCLu-DLBCL/CHL without mediastinal disease.
Three cases are in the peripheral lymph nodes as a primary site and a case is in the
extranodal region. All of the cases were histologically similar to nodular sclerosis
CHL and the tumor cells were positive for CD30 and CD20. We examined the cases
according to WHO 2008 criteria and discussed in the light of the literature.
Keywords: Gray-zone lymphoma, immunohistochemistry, in situ hybridization (ISH)
Figure 1. CD10 expression on B-lymphoid cells.
follicular lymphoma has been reported rarely in the English literature (less than 20
cases) but even more rarely they were reported as being negative for t(14;18) or
presenting with extranodal involvement.
[PP-LYMP-104]
Figure 2. CD5 expression on T cells (strong) and B-lymphoid cells (weak to
moderate).
[PP-LYMP-106]
TRANSLOCATION T(14;18) NEGATIVE LOW-GRADE
FOLLICULAR LYMPHOMA WITH CONCOMITANT EXPRESSION
OF CD5 AND CD10
Chad Ellermeier, Diana Olguta Treaba
Department of Pathology and Laboratory Medicine, Lifespan Academic Medical Center, The Alpert
School of Medicine at Brown University, Providence, Rhode Island
Introduction: Concomitant co-expression of CD5 and CD10 by B-cell lymphomas
is rarely seen with an estimated incidence of 0.4% of the B-cell lymphomas in the
series of Henry Y et al, 2003.
Case: We present a 79-year old female patient who noticed increasing difficulty in
swallowing and a direct fiberoptic examination identified right tonsillar hypertrophy
and a base of tongue lesion.
Results: A right tonsillectomy and associated biopsy of the tongue lesion were remarkable for effacement of the normal architecture by variable in size ill-defined
lymphoid follicles composed predominately of small lymphoid cells, centrocytes and
admixed scattered centroblasts (<15 centroblasts/hpf). By immunohistochemistry,
the neoplastic lymphoid follicles were CD20 and PAX5 positive, had strong co-expression of bcl2 and germinal center markers (bcl6 and CD10) and had also weak
to moderate CD5 co-expression. They were CD43 positive and negative for CD23,
Cyclin D1 and MUM-1. The neoplastic lymphoid follicles were centered by variably
disrupted CD23 positive follicular dendritic meshworks and their proliferation rate
ranged from 10-15% to 20-30% as identified by the immunoreactivity to the MIB-1
antibody. FISH studies were also performed and were reported negative for t(14;18)
and also negative for bcl6 rearrangements.
Conclusions: A diagnosis of low-grade follicular lymphoma with concomitant coexpression of CD5 and CD10 was rendered. The bone marrow examination was
negative for involvement by lymphoma. A CT of the neck, chest and abdomen did
not detect any regional or metastatic disease. For her stage IE low-grade follicular
lymphoma the patient was treated with local radiation therapy and is still in remission18 months since her diagnosis. The unusual co-expression of CD5 positivity in
Figure 3. Tonsil, hematoxylin and eosin stain, 50x.
[PP-LYMP-107]
CLINICOPATHOLOGICAL FEATURES OF PRIMARY BONE
LYMPHOMA, A SINGLE- INSTITUTION STUDY
Fatemeh Varshoee Tabrizi1, Mohammad Reza Ghavam Nasiri1,
Amir Aledavood1, Bahram Memar2, Kamran Ghafarzadegan2
1
Department of Radiation Oncology, Cancer Research Center, Mashhad University of Medical
Sciences, Mashhad, Iran
2
Department of Pathology, Mashhad University of Medical Sciences, Mashhad, Iran
Background: Primary bone lymphoma (PBL) is a rare disease and distinct clinicopathological entity. The optimal treatment strategy is still unclear. Because of rarity
of PBL, we report our institute experience in PBL clinicopathological feature and
treatment results.
Patients and Methods: 28 patients diagnosed with PBL were referred to Omid
Hospital (C.R.C), between March 2001 and February 2009. Immunophenotype
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127
Table 1. Patients’ demographic characteristics
Results: 14-28 patients with PBL were analyzed retrospectively.17 patients (60.7%)
were male and 11(39.3%) female with a median age of 41 years (range: 11-79).long
bones were the most primarily site of involvement (71%). 26 (93%) patients had
DLBCL and 2(7%) with small lymphoblastic lymphoma. One(3%) patient received
radiation alone, 18(66%) cases received combined modality (chemotherapy + radiotherapy) and 8(30%) received only chemotherapy during their treatment period.
Parameter
The median follow up was 18 months (range: 1-82).
Sex
Mean of DFS was 51 months (range: 37-66).
Male
17 (60)
Overall survival was 54 months (range: 40-68).
Female
11 (40)
Diffuse large B cell lymphoma (DLBCL) and 2 (7%) with small lymphoblastic lymphoma.OS was significantly better in the chemoradiotherapy group (64 versus 27
months) p=0.014.
Male:female ratio
DFS was also significantly better in combined modality arm (64 versus 21 months)
p=0.003.
Table 2. Patients’ clinical characteristics
number%
Patients
28
Median age (year)
41
<60 year
23 (82)
>60 year
5 (18)
Parameter
Conclusion: In spite of small number of patients reported in this study, combined
modality treatment (chemotherapy and radiotherapy) is useful as an effective treatment strategy in PBL.
1.5/1
Number %
Stage
Keywords: primary bone lymphoma, chemotherapy, radiotherapy
I
17 (60)
II
4 (15)
IV
B symptoms
7 (25)
15 (54)
LDH
th
studies on 16 out of 28 pathological blocks were performed. We analyzed disease
free survival (DFS) and overall survival (OS).
Normal
6 (30)
Elevated
14 (70)
IPI score
Low
6 (30)
Low-intermediate
7 (35)
High
7 (35)
Treatment
Figure 1. DFS curves comparing Chemoradiotherapy(CRT) (1) versus
chemotherapy alone.
Chemotherapy
8 (30)
Chemotherapy + Radiotherapy
18 (66)
LDH, lactate dehydrogenase; IPI, international prognostic index; B symptoms (fever>=38•c, night
sweating, weight loss>=10 kg in six months)
[PP-LYMP-108]
PRIMARY GASTROINTESTINAL LYMPHOMA
Soodabeh Shahidsales1, Amir Aledavood1,
Mohammad Reza Ghavam Nasiri1, Bahram Memar2,
Hamid Reza Raziee1, Kamran Ghafarzadegan2, Samira Mohtashami1
1
Cancer Research Center, Faculty of Medicine, Mashhad University of Medical Sciences,
Mashhad, Iran
Department of Pathology, Omid Hospital, Mashhad University of Medical Sciences, Mashhad, Iran
2
Figure 2. OS curves comparing Chemoradiotherapy(CRT) (1) versus
chemotherapy alone.
Background: extranodal lymphoma may arise anywhere outside lymph nodes, gastrointestinal (GI) tract is the most frequent site of extranodal involvement by nonHodgkin’s lymphoma.We reviewed the clinicopathological features and treatment
results of patients with primary GI lymphoma.
Materials-Methods: A total number of 30 cases with primary GI lymphoma were
included in this study. Patients referred to the Radiation Oncology Department of
Omid Hospital (Mashhad, Iran) during a 5-year period (2006-11). Clinical, paraclinical, and radiological data was collected from medical records of the patients. The
patients’ clinical staging was determined according to the Ann Arbor classification.
Figure 3. Overall survival curve.
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Results: Out of the 30 patients with primary GI lymphoma in the study, 12 were
female (40%) and 18 were male (60%) (male to female ratio: 3/2). B symptoms were
present in 27 patients (90%). The mean age of patients was 50 ± 16.9 years (range:
15-79 years). The characteristics of the patients and results of laboratory tests are
summarized in Tables 1 and 2, respectively. Lactate dehydrogenase (LDH), as an
important prognostic factor in non-Hodgkin’s lymphoma,was elevated in 9 patients
(32.1%). The most common primary site was stomach in 14 cases (46.7%). Other
common sites included small intestine and colon each in 8 patients (26.7%). All patients had histopathologically proven non-Hodgkin’s lymphoma. The most common
Table 2. Results of Laboratory tests of patients with primary GI Lymphoma
Variables
International Prognostic Index (IPI) score and final condition of patients are presented in Figures 1 and 2,respectively. In addition, 28 patients (93.3%) received
chemotherapy with cyclophosphamide, vincristine, doxorubicin, prednisolone (CHOP
regimen). The median course of chemotherapy was 6 courses. Moreover, 8 patients
(26.7%) received radiotherapy with cobalt 60. The median follow-up time was 26
months. The overall 5-year survival rate was 53% and the median survival time
was 60 months.
Conclusion: Primary GI lymphoma is commonly seen in stomach and small intestine
and mostly is DLBCL or mucosa associated lymphatic tissue (MALT) lymphoma.
Frequency (%)
Lactate dehydrogenase
(LDH)
<=500
>500
19 (67.9%)
9 (32.1%)
Erythrocyte sedimentation rate
(ESR)
<=50
>50
20 (87%)
3 (13%)
Hemoglobin
(Hb)
<10
10-12
>12
7 (23.3%)
13 (43.3%)
10 (33.3%)
<=100.000
>100.000
1(3.3%)
28 (93.3%)
<=3000
>3000
2 (6.7%)
27 (90%)
Aspartate aminotransferase
(SGOP)
<=40
>40
22 (73.3%)
7 (23.3%)
Alanine aminotransferase
(SGPT)
<=40
>40
25 (83.3%)
4 (13.3%)
Platelets
(PLT)
Keywords: Primary Gastrointestinal Lymphoma, Diffuse Large B-Cell Lymphoma,
Extra nodal Lymphoma
White blood cell count
(WBC)
[PP-LYMP-109]
ANGIOIMMUNOBLASTIC T-CELL LYMPHOMA PRESENTING
WITH EBV POSITIVE DIFFUSE LARGE B-CELL LYMPHOMA OF
THE ELDERLY
Gitte Wooler1, Peter Nørgaard1, Tim S. Poulsen1, Michael Pedersen2,
Xiangrong Zhao3, Stefania Pittaluga3, Elaine S. Jaffe3,
Francesco d’Amore4, Signe Ledou Nielsen1
Figure 1. IPI score of patients with primary GI lymphoma.
histologic subtype was diffuse large B-cell lymphoma (DLBL) in 16 patients (53.3%).
According to Ann Arbor staging system, 43.3% of patients were in stage I, 36.6% in
stage II, 6.6 % in stage III, and 13.3% in stage IV. The
1
Department of Pathology, Herlev University Hospital, DK-2730 Herlev, Denmark
Department of Haematology, Herlev University Hospital, DK-2730 Herlev, Denmark
Lab. of Pathology, Center for Cancer Research, National Cancer Institute, National Institutes of
Health, Bethesda, MD20892, USA
4
Department of Hematology, Aarhus University Hospital, Aarhus, Denmark
2
3
Angioimmunoblastic T-cell lymphoma (AITL), one of the most common types of
peripheral T-cell lymphoma, is often associated with immunodeficiency. In rare
cases this leads to secondary Ebstein-Barr virus (EBV) driven large B-cell lymphoma
(LBCL). We report a case of AITL with the unusual presentation of EBV-positive diffuse LBCL, one year prior to clinical manifestations from AITL.
Figure 2. Final condition of patients with primary GI lymphoma in Omid hospital
(Mashhad/ IRAN).
Table 1. Characteristic features of patients with primary GI Lymphoma
Variables
Value
Frequency (%)
Age
53.44±15.8
44.75±17.9
sex
male
Female
18 (60%)
12 (40%)
Primary site
Intestine
Colon
Stomach
8 (26.7%)
8 (26.7%)
14 (46.7%)
B symptoms
performance
International Prognostic
Index
(IPI score)
27 (90%)
0
1
2
3
4
1 (3.3%)
7 (23.3%)
11 (36.7%)
10 (33.3%)
1 (3.3%)
low
Low intermediate
High intermediate
high
13 (46.4%)
10 (35.7%)
4 (14.3%)
1 (3.6%)
A 64 year old, previous healthy, woman was diagnosed with EBV-positive DLBCL of
the elderly in a cervical lymph node showing strongly positive EBER-ISH and clonally rearranged immunoglobulin heavy chain (IgH) genes. The lymph node showed
no signs of AITL. Clinically the patient had stage IVB (lymphomas on the neck, thorax and abdomen, bone marrow negative), IPI=III, increased lactate dehydrogenase
(LDH), and high numbers of EBV-copies. Immunoglobulins were in the normal range.
The patient received 6 cycles of R-CHOP chemotherapy which resulted in complete
remission (PET/CT scan) although LDH remained slightly increased. One year later
clinical control showed relapse with enlarged, PET-positive lymph nodes. The patient was however well and LDH had spontaneously normalized. AITL was diagnosed
in an excised axillary lymph node, based on typical histological and immunohistochemical characteristics and clonally rearranged T-cell receptor (TCR) genes. The
patient received high dose chemotherapy (BEAM) followed by successful bone marrow stem cell transplantation. However four months later the patient experienced a
rapid relapse with generalized lymphadenopathy and clinical deterioration. Axillary
lymph node excisions showed AITL with clonal TCR gene rearrangements identical
to previous analyses and polyclonal IgH gene rearrangements. The bone marrow
was strongly infiltrated with EBV-negative DLBCL with no signs of AITL (confirmed
by flow cytometry). The patient was discharged home with palliative care.
No previous cases have been reported of subclinical AITL presenting with EBVpositive DLBCL. While AITL is sometimes associated with EBV-positive B-cell proliferations in this case we cannot conclude whether these were two distinct processes or there was subclinical AITL possibly predisposing the observed EBV-positive
DLBCL. We suggest that in patients with EBV-positive DLBCL of the elderly, clinicians
should be aware of the possibility of predisposing, subclinical AITL.
Keywords: Angioimmunoblastic TCL, EBV positive DLBCL
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129
th
[PP-LYMP-110]
WITHDRAWN
[PP-LYMP-111]
UNIQUE COMPOSITE FOLLICULAR LYMPHOMA AND MANTLE
CELL LYMPHOMA WITH INTERMINGLED TUMOR CELLS IN
THEIR APPROPRIATE MORPHOLOGIC COMPARTMENTS
Kristina Fasig1, Vladimir D. Kravtsov 2nd1, Annette S. Kim2,
David Head2
1
2
PathGroup, Brentwood, Tennessee, USA
Department of Pathology, Vanderbilt University Medical Center, Nashville, Tennessee, USA
History: The patient is a 55 year old male with no significant prior medical history,
who presented with an enlarged left inguinal lymph node of at least several months
duration. An ultrasound showed multiple enlarged inguinal nodes, the largest measuring 3 cm. in greatest dimension. A biopsy was performed.
Morphology: Review of H&E stained material revealed increased, closely spaced,
regular follicular centers with loss of polarity, relatively uniform cellular composition,
and lack of tingible body macrophages, with expanded mantle zones. Interfollicular
and subcapsular regions of the node showed an infiltrate of uniform slightly enlarged
lymphocytes with a diffuse pattern, with morphology similar to those comprising
the mantle zones around follicles. Morphologic assessment was thus suspicious for
presence of two distinct processes, one with a pattern of follicular lymphoma, the
second suspicious for mantle cell lymphoma.(Fig.1) Immunohistochemistry demonstrated the follicles were composed of cells that were CD20+/CD10+/BCL6+/
BCL2+/cyclin D1-,(Fig.2a,c) while smaller extrafollicular cells in mantle zones surrounding these follicles and elsewhere in the node were CD20+/CD10-/BCL6-/
BCL2+/cyclin D1+.(Fig.2b,d)
Figure 2. Immunohistochemical stains of serial sections of lymph node, 4x. A.
CD10; B. BCL2; C. CD5; D. Cyclin D1. A and B show follicular centres staining for
CD10 and BCL2, characteristic of follicular lymphoma; note that mantle cells also
stain positively with BCL2. C and D show mantle cells staining with CD5 and Cyclin
D1, characteristic of mantle cell lymphoma.
Flow cytometry: Flow cytometry demonstrated 2 populations of clonal B-cells. The
first was CD19+/CD20+/CD10+/monotypic lambda,(Fig.3a) the second CD19+/
CD20+/CD5+/ CD23-/monotypic kappa.(Fig.3b)
FISH: FISH analysis revealed an IgH/BCL2 rearrangement in 17% of cells analyzed,
and a CCND1/IgH rearrangement in 38% of cells.
Diagnosis: Lymph Node, left groin: Composite Small B-cell Lymphoma With 2
Distinct Processes, Mantle Cell Lymphoma (approximately 70%) and Follicular
Lymphoma, Follicular Pattern, WHO Grade 2/3 (approximately 30%)
Comment: This case is unusual, perhaps unique, in that it shows two newly diagnosed distinct lymphoproliferative processes superimposed on one another in the
diagnostic biopsy, within their normal and intermingled lymph node compartments.
The two processes are apparently clonally unrelated and likely represent the spontaneous and separate acquisition of their respective disease-driving translocations
that are neoplastically populating the same lymph node(s). This is distinct from the
typical collision lymphoma where a lymph node might have two geographically
separated lymphomatous processes. We are aware of a single report of a similar
case (Liu et al, Am J Clin Pathol 2014;141;737-741, which showed these same
two diseases admixed in a single lymph node, but with only the pattern of follicular
lymphoma.
Keywords: unique composite lymphoma
Figure 1. Lymph node section (H&E) 4x, insert 20x, showing closely packed
regular follicles with loss of polarity (fine arrows) with surrounding prominent mantle
zones (course arrows).
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Figure 3. Flow cytometry histograms gating on CD19+ cells. A. CD10+ B-cells
are kappa negative. B. CD5+ cells are kappa positive.
[PP-LYMP-112]
T CELL LYMPHOBLASTIC LYMPHOMA BREAST INVOLVEMENT:
A CASE REPORT
Figen Atalay1, Semsi Yildiz2, Oyku Gulmez3, Hakan Altay3,
Ebru Demiralay2
1
Bașkent University school of medicine, Department of Hematology, İstanbul
Bașkent University school of medicine, Department of Pathology, İstanbul
3
Bașkent University school of medicine, Department of Cardiology, İstanbul
2
Introduction: T lymphoblastic cells derived from T cell precursors. T lymphoblastic
cells can produce T cell lymphoblastic lymphoma (T-LBL). Difference between T-LBL
and T- Acute lymphoblastic leukemia ( T-ALL) defined as infiltration of bone marrow
above 25% in the World Health Organization (WHO) classification in 2008 (1). T-LBL
infiltration predominantly is seen at lymph nodes, central nervous system, testes.
but occasionally extramedullary infiltration can be seen (2). We reported here a patient who has T-LBL was presenting with breast masses and pericardial effusion.
Case: A 49 years old caucasian woman, who was no prior medical history, admitted
to cardiology clinic because of chest pain and dsypnea in februrary 2014. She had
no prior known medical history. Massive pericardial effusion was determined in
examination of transthorasic echocardiography. Pericardiocentesis was performed
to her. The pericardial fluid was found to be exudative and atypical lymphocyte predominancy. At the same time bilaterally breast masses were palpated in physical
examination of her and bilaterally multifocal breast nodulary lesions were found in
breast ultrasound. Mediastinal, paraaortic lymph node enlargements were detected
in thoracal and abdominal computurized tomography. There was no abnormality in
her complete blood count, peripheral blood film and biochemical laboratory tests.
Bone marrow biopsy performed to her and %20 T lymphoblastic cell infiltration
was shawn. The Iymphoblasts in T-ALL/ LBL are usually TdTpositive and variably
express CDla, CD2, CD3, C04, CD5, C07 and CD8. Of these,CD7 and cytoplasmic
CD3 are most often positive, but of these only CD3 is considered lineage specific.
Conclusion: Extramedullary T-LBL infiltration of the breast is extremely rare. We
present here a patient who found that infiltration of breast with lymphoma cells
while diagnostic and staging procedures apllying.to her.
Keywords: T cell lymphoblastic lymphoma, breast
[PP-LYMP-113]
NON-HODGKIN LYMPHOMA WITH RENAL INVOLVEMENT
Seyma Ozkanlı1, Ebru Zemheri1, Erman Ozturk2, Bengu Cobanoglu1,
Ali Bakan3, Tulay Zenginkinet1, Abdullah Aydın1, Tulay Tecimer4
1
Department of Pathology, Medeniyet University Göztepe Training and Research Hospital, İstanbul,
Turkey
Department of Hematology, Medeniyet University Göztepe Training and Research Hospital, İstanbul,
Turkey
3
Department of Nephrology, Medeniyet University Göztepe Training and Research Hospital, İstanbul,
Turkey
4
Department of Pathology, Acıbadem University, Faculty of Medicine, İstanbul, Turkey
2
BLASTIC PLASMACYTOID DENDRITIC CELL NEOPLASM:
CASE REPORT
Mehmet Dogan1, Aydan Kilicarslan2, Sibel Orhun Yavuz2,
Nuran Sungu2, Gulsen Akoglu3
1
Dr. Abdurrahman Yurtaslan Ankara Oncology Education and Research Hospital, Department of
2
3
Dermatology, Ankara, Turkey
Background: Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is also known
as CD4+ / CD56+ hematodermic neoplasm. It is a rare and highly aggressive hematopoietic malignancy. According to World Health Organization (WHO) classification
system, this tumor was originally termed as blastic natural killer-cell lymphoma
because of its distinct cytology and CD56 expression. In addition the latest studies
directed us using the term “plasmacytoid dendritic cell leukemia/lymphoma” may
be more appropriate. Clinically skin is the most frequently involved site. But lymph
node and bone marrow involvement can also be seen.
Case: 65 years old male patient presented with a firm mass in the forehead skin. The
lesion had gradually grown for six months. The lesion was 5x4 cm in diameter and
1 cm elevated from skin. Surface of the lesion was red to violet in color. Excisional
biopsy of the lesion was characterized by a non-epidermotropic, dermal and subepidermal infiltration of homogeneous medium-sized cell resembling lymphoblasts
or myeloblasts. Immunohistochemical stains showed that tumor cells were diffusely
positive with CD4 and CD56. Negative markers are included CD3, CD20, pax-5,
CD30, myeloperoxidase and TdT. Ki-67 proliferation index was about %80 of the
cells. Neither T nor B cell receptor gene rearrangements were detected. There was
no involvement in bone marrow biopsy. In follow-up new lesions observed on trunk
and upper extremity skin which were histologically confirmed as same lesion with
primary lesion. The lesions regressed remarkably after chemotherapy was started.
Discussion: Lymphoblastic lymphoma is an aggrressive lymphoma which was
classified in WHO 2008 (1). T-LBL usually presents with mediastinal lymph node
enlargements, But occasionally it present with extramedullary infiltration. NonHodgkin lymphoma’s (NHL) of the breast is very rare (1.7-2.2% of all NHL) and
originate from usually B cell lineage (3). T-cell subtype lymphoma of breast is uncommon.Only 10 cases was reported in literature (4-9). Our case is the second case
in the literature which was infiltrated the breast with T- LBL cells. ALL like regimens
recommended that the patients who have T-LBL (10). We gave her CALGB CT protocole and her treatment protocole is still continue.
[PP-LYMP-114]
CD4 and CD8 are frequently co-expressed on the blasts, and CD10 may be positive.
These latter phenotypes are not specific for T-ALL as CD4 and CD8 double positivity In addihon to TdT, the most specific markers to indicate the precursor nature of
TIymphoblasts are CD99, CD34 andCDla. Expression of CD117, or c-Kit, is thought
to be a relatively specific marker for myeloid differentiation; however, CD117 is
occasionally seen in T-cell ALL as well. CD117 expression is observed in very early
normal T-cell precursors. Same immunocytochemistrical staining features were
seen in breast biopsy. She accepted as T-LBL Ann-Arbor stage IVB and CALGB 8811
chemotherapy (CT) treatment protocole was applied to her. After one week of the
CT her pericardial effusion,dyspnea and chest pain were resolved. She is in early
intensification phase according to the CT protocole.
Discussion: BPDCN newly defined entity which is originating from CD123 positive
plasmocytoid dendritic cells. Awareness of such a tumor and new immunohistochemical markers (CD123, CD303, TCL1) may help the diagnosis of BPDCN.
Keywords: dendritic cell, neoplasm, plasmacytoid
Renal involvement in non-Hodgkin lymphoma (NHL) was previously reported, including glomerulonephritis (GN), acute kidney injury, and lymphoma infiltrating the
kidney parenchyma. Previous studies showed that up to 10% of the patients with
NHL and lymphocytic leukemia may have kidney injury. On the other hand, a limited
number of cases of GN have been described in patients with NHL demonstrated by
renal biopsy in the literature.
A 59 year-old male patient was presented as severe abdominal pain. On physical examination, unilateral 2x3 cm lymphadenopathy was detected. At blood
and urine tests, WBC:7200/mm3; Hb:7,6gr; Htc:22%; MCV:96fl; Plt:227.000/
mm3; Creatinin:1,8mg/dl; total protein:4,4gr; Albumin:2,5 gr; ESR:66mm/h and
5g/day proteinuria was detected. Subsequent bone marrow biopsy revealed a
low grade small B cell lymphoma with CD5 positive, CD20 positive, bcl-2 positive. Membranoproliferative glomerulonephritis –like pattern (MPGN) and infiltration of low grade small B cell lymphoma with CD5 positive, CD20 positive, bcl-2
positive was diagnosed in renal biopsy. There was seen small B cell lymphoma
with plasmositoid differentiation, kappa monoclonal in biopsy of lymph node. The
case was evaluated as a low-grade B-cell lymphoma with kidney and bone marrow
involvement.
NHL might be first diagnosed by renal biopsies for evaluation of kidney injury or proteinuria. CLL/SLL were the most common types of NHL associated with renal injury,
and the most common pattern of glomerular lesion was MPGN-like pattern. Renal
biopsy is necessary to establish the underlying cause of renal involvement in NHL.
Keywords: non-Hodgkin lymphoma (NHL), renal biopsy, kidney involvement
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SOCIAL PROGRAM
SOCIAL PROGRAM FOR PARTICIPANTS
Participants will have the opportunity of enjoying a diverse programme of activities, such as the Welcome
Reception, Speakers Dinner on Bosphorus and the Gala Dinner on Suada reflecting the rich multicultural
character of İstanbul.
WELCOME COCKTAIL
Will be held in “Hilton Hotel-Ball Room “(Congress venue, -1 Floor)
Date
Time
: October 18, 2014, Saturday
: 18:30 – 20:00
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S O C IAL PRO G RA M
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Will be held in “Suada”
Gala Dinner Fee Only: 115 €* (for unregistered participants, except accompanying and Pack Registration)
*Child: under 12 years (0-6 free, 7-12 full charge)
Date
Dinner Time
: October 20, 2014, Monday
: 20:00 – 23:00
Bus Transfer from Hilton İstanbul to Kuruçeşme Port
Time : 18:45
Place : Hilton Hotel Convention Center (Congress Venue)
Transfer from Kuruçeşme Port to Hilton İstanbul
Time : 23:00
Place : Kuruçeşme Port
The island at the Kuruçeşme district of Bosphorus,
which is composed of several big stone blocks 165
meter away from the shore was given as a present
to the Serkis Kalfa, the chief architect of the palace
in 1872 by Sultan Abdulaziz, the ruler of the Ottoman
Empire. Having built a three storey pavilion in this
island, Serkis Kalfa moved here.
GALA DINNER
The worldwide famous painter Ayvazovski, invited
by the Sultan Abdulaziz, had been a guest to Serkis
Kalfa at the Kuruçeşme Island in 1874 and was
introduced to the sultan.
Ayvazovski painted the pictures of Dolmabahçe
Palace ordered to be drawn by the Sultan Abdulaziz
at this island. The island was rented out to the
‘’Şirket-i Hayriye Ferry Enterprise’’ by the heirs of
Serkis Bey after the 2nd World War and it was used
as a coal depot for long years.
This island was called as ‘’The island of Serkis Bey’’ up to the 1st World War years and was known to be
‘’A corner of heaven’’ of the time. Serkis Kalfa (1835-1899), who continued to render service as the chief
architect of the palace subsequent to the death of Sultan Abdulaziz and during the reign of Abdulhamit 2nd,
lived at this island until his death
The island was bought by Galatasaray Sports Club in 1957 and it was turned into a place of social facilities.
You will be required to show your invitation cards to attend the Gala Dinner.
REMINDER
Please change your Gala Dinner Ticket at the Registration Desk for your invitation card latest by
Saturday, October 18, 2014 – 16.00
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İstanbul RAIL LINES NETWORK MAP
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İstanbul
İstanbul, being the only city in the world built on two
continents, has been determined by its vital strategic
location and enchanting beauty. İstanbul stretches
along the two shores of the Bosphorus that links the
Sea of Marmara with the Aegean Sea in the South
and with the Black Sea in the North.
It has long been coveted by powerful empires, and
served as capital to Roman, Byzantine and Ottoman
Empires, and currently enjoys being the largest city of
the Republic of Turkey, with a population of 14 million.
İstanbul is also the capital of art and culture with a
rich tradition in opera and ballet, theatres performing
Turkish and foreign plays, concerts, art exhibitions,
festivals, auctions, conferences and of course unique
museums. The city also boasts the country’s largest
and finest universities.
As an imperial capital for 1500 years, İstanbul has
acquired a highly original personality. At every turn
in the city you are faced with Roman, Byzantine and
Ottoman palaces, mosques, churches, monasteries,
monuments, walls and ruins.
TOURIST INFORMATION
Yet İstanbul is not a city living in its past. It is a vibrant,
modern and future-oriented metropolis. Bazaars and
ultra-modern department stores, street vendors and
stock-brokers, old crumbling buildings and skyscrapers, horse-drawn carts and luxurious limousines, fine
dining restaurants, modern cafes and street corner food
shops coexist and this mix gives the city a multi-faceted
outlook and flavor. You could taste the pre-modern,
modern, and post-modern side by side in this city.
All about İstanbul (Turkish, English):
http://english.İstanbul.com/
Banking & Exchange Facilities
Foreign money can be changed at banks during business days (09.00-17.00 Monday-Friday) as well as at
hotels, at the airport and in exchange offices. Exchange rates are set daily by the Central Bank. All major
credit cards are accepted in most hotels, restaurants and shops. Automated bank machines are available at
many points throughout the city and at the airport.
Climate & Clothing
The start of October is the best time in Spring. Daytime temperatures are in the range of 18–26ºC and
evening temperatures are 13–17ºC. We recommend you bring the clothes regarding the weather conditions
and a raincoat as rain showers occur.
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The official meeting currency is Euro, fees for registration, hotel accommodation and tours need to be paid
in Euro. The exchange rate for Turkish Lira can be learned from the web site of Turkish Central Bank’s web
site (www.tcmb.gov.tr).
Credit Cards
International credit cards are accepted in cash dispensers, hotels, restaurants and most shops, as well as
car rental agencies. The most common credit cards are VISA, Euro Card, and MasterCard.
Driving License
International Driving Licenses are recognized throughout Turkey. Car rental companies ask for a valid
driving license.
Electricity
The electrical power supply in Turkiye has 220 volts.
Emergency Telephone Numbers
Ambulance
Fire Dept
Police
112
110
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Currency
Health
Turkiye has a high standard of hygiene and doctors and dentists are highly trained and hospitals are well
equipped. In the event of illness, hotel staff can arrange a doctor for you.
Insurance
Registration fees do not include insurance of any kind. It is strongly recommended that delegates take out
adequate travel and health insurance prior to commencement of travel. Further information can be obtained
from your travel agent.
Mobile Phone
There are 3 GSM operators in Turkey being as Turkcell, Avea and Vodafone. All of them are daughters or
partners of international companies and have roaming agreements with network operators of all participating
countries. There are two wave bands for mobile phones in Turkey: 900 and 1800 MHz. Please check with
your provider regarding roaming costs and wave bands. Local GSM operators have prepaid lines, which can
be provided at newspaper booths and groceries. Please check that the prepaid cards are in accordance with
your line. Additionally, there will be renting possibilities of mobile phones with lines.
Post Office
Post offices are indicated PTT (Post, Telegraph, and Telephone) throughout the city.
The central Post office is open Monday through Saturday from 8:00 to 21:00, Sunday from 9:00 to 19:00.
Smaller ones are open Monday through Friday between 8:30 and 17:00.
Hotel concierges also take the mail.
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There is a wide choice of restaurants in İstanbul offering everything from excellent national cuisine to first
class international dishes. Traditional Turkish cuisine is famous for its many specialties prepared with fresh
vegetables. There are lots of quality restaurants and fast-food shops in the vicinity of the hotels reserved
for this meeting.
Security
Based on data provided by well known statistical institutes worldwide, İstanbul, among other metropolises,
is regarded as one of the few cities with a low rate of crime. In the Hilton Hotel Convention Center security
staffs are on duty using the newest technological security systems.
Shopping
One of the most enjoyable parts of a trip to İstanbul is shopping for the rich variety of Turkish crafts. İstanbul
is a shopping paradise with its Covered Bazaar as well as the modern malls. Turkish carpets, kilims, suede
and leatherwear as well as cotton shirts and clothing are the best buys here. Copper, silver, brassware and
jewelers are also choices for buying, along with works of ceramics and the famous Turkish meerschaum.
Bargaining over the sale price with shop traders is expected.
Smoking
Smoking is not permitted inside of restaurants, public service vehicles, museums, libraries, lifts, theatres,
cinemas, supermarkets, department stores and government offices.
Offenders can be fined up to 100 Turkish Liras.
Inline with the efforts to improve the nightlife experience for all, there are smoking restrictions on entertainment
outlets. Smoking is no longer allowed in all pubs, discos, karaokes, bars and other public nightspots unless
within approved smoking rooms /corners.
Effective 1 January 2009, smoking is prohibited at:
• Indoor public places e.g. shops, shopping/neighborhood centers factories, offices regardless of whether
they are air conditioned.
• Lift and hotel lobbies.
• Within 5 m of entrances and exits to indoor area of buildings and facilities where smoking is prohibited.
• Playground and exercise areas.
• Markets.
• Ferry terminals.
Restaurants & Turkish Cuisine
Taxis
Available at taxi stands or hailed on the street. All are yellow and have meters.
Telephones
Pay phones are available at the meeting venue as well as in the city.
Time Differences
Turkey is 2 hours ahead of Greenwich Mean Time and 7 hours ahead of Eastern Standard Time.
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Tipping
Tipping is widespread or regulated in Turkiye as it is in other parts of the world. But tipping depends on your
choice, a reward for service. It is customary to tip hotel porters, and a gratuity of about 5-10% is usual in
restaurants if good service is received. Service charge can be added to bill at some hotel and restaurants.
If service charge is added to the bill, you do not need to tip for them.
Transportation
İstanbul Atatürk International Airport is an important destination and transfer point for many international
flights from all over the world.
Transfer information from the Airport to City Center by taxi, train or bus:
http://www.Atatürkairport.com/en-EN/Transportation/Pages/AirportTrasnportation.aspx
Airlines
TURKISH AIRLINES
BRITISH AIRWAYS
SABENA AIRLINES
AIR FRANCE
KLM
LUFTHANSA AIRLINES
SWISS AIRLINES
FINNAIR
LOT POLISH AIRLINES
ALITALIA
MALEV
SCANDINAVIAN AIRLINES
TAP-AIR PORTUGAL
ROMANIA AIRLINES
OLIYMPIC AIRLINES
AUSTRIAN
IBERIA
AEROFLOT
AIRLINES EASYJET
SUN EXPRESS
(Sun Express has very reasonable rates from Germany, Switzerland, Austria to İstanbul.)
Airport İstanbul – Atatürk International Airport
34149 Yeşilköy, İstanbul, Turkey
Phone: +90 212 465 55 55
Fax: +90 212 465 50 50
e-mail: info@tav.aero
www.Atatürkairport.com
Atatürk Airport is located in Yeşilköy, in the european side of İstanbul. Airports distance is 28 km to downtown,
30 km to Harbiye Military Museum and 4 - 4.5 km to sea shore.
Sabiha Gökçen International Airport
34912 Pendik, İstanbul, Turkey
Phone: +90 216 585 50 00
Fax : +90 216 585 51 14
e-mail : heas@sgairport.com
Sabiha Gökçen Airport is located in Pendik/Kurtköy, in the Asian side of İstanbul. Airports distance is 40
km to Kadõköy, 12 km to Pendik, 50 km to Taksim and 60 km to Harbiye Military Museum.
It has a really convenient traffic in terms of transportation with its 1.5 km connection to the TEM motorway.
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www.goturkey.com
Official tourism portal of Turkey (Turkish, English, German, French)
www.mfa.gov.tr
Official portal of the Ministry of Foreign Affairs (English, French, Arabic)
www.allaboutturkey.com
An award winning private portal of tourism (English, French, German, Spanish, Russian, Arabic, Italian,
Indian)
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TURKEY
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O F F IC IA L C A RRI ER
SAVE UP TO 20% ON TRAVEL WITH THE STAR ALLIANCE™ NETWORK
The Star Alliance member airlines are pleased to be appointed as the Official Airline
Network for 17th Meeting of the European Association for Haematopathology.
To obtain the Star Alliance Convention Plus discounts please follow the below steps to
access the Convention Plus online booking tool:
• Visit www.staralliance.com/en/business-solutions/conventions-plus/delegates/
• Under “Delegates login” enter conventions code TK03S14.
• The online booking tool opens in a separate window*
*Should the online booking tool not open, please ensure that your Pop-Up blocker is disabled.
Registered participants plus one accompanying person travelling to the event can qualify
for a discount of up to 20%, depending on fare and class of travel booked.
The participating airlines for this event are: Ethiopian Airlines, Air Canada, Air New
Zealand, ANA, Austrian Airlines, Lufthansa, Scandinavian Airlines, Singapore Airlines,
THAI, United, Asiana Airlines, LOT Polish Airlines, TAP Portugal, South African Airways,
Air China, Turkish Airlines, EgyptAir, Adria Airways, Croatia Airlines, Brussels Airlines,
Aegean Airlines, US Airways.
Discounts are offered on most published business and economy class fares, excluding
website/internet fares, senior and youth fares, group fares and Round the World fares.
Please not: For travel to/from Japan and New Zealand special fares or discounts my be
offered by the participating airlines on their own network. To obtain these special fares
or discounts and for booking office information please visit www.staralliance.com/en/
business-solutions/conventions-plus/delegates/ and:
• Click on “Conventions Plus Booking Contacts” and enter the conventions code
TK03S14.
• Choose one f the participating airlines listed
• Call the respective reservation contact listed and quote the conventions code TK03S14
when requesting the special ticket
When making your travel plans please present confirmation of your registration or proof of
attendance for the Event/Convention.
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İSTANBUL TOURS INFORMATION
TR-01 THE IMPERIAL TOUR (Daily Morning Tour)
We begin our tour of the Sultanahmet
district, the heart of old İstanbul, at Hagia
Sophia*. Built by the Emperor Justinian
in the early 6th century AD and designed
by Anthemius of Tralles and Isodore of
Miletus, the church is one of the marvels
of world architecture. Converted into a
mosque in 1453, it is now a museum. Its
massive dome still dominates the skyline of
old İstanbul. It is also famed for its mosaics, including glittering portraits of emperors and empresses and a poignant Virgin and Child. Next we visit the Blue Mosque which takes its name
from the exquisite tiles adorning its interior. Built by Sultan Ahmet Ist in the early 17th century and designed
by a pupil of Sinan, the greatest of Ottoman architects, it is the only imperial mosque with six minarets. Its
courtyard is especially grand. The Hippodrome, the stadium of ancient Byzantium, held 100,000 spectators
and featured objects from all corners of the empire. Of these, an Egyptian obelisk and a bronze sculpture of
three entwined serpents from Delphi survive. The Grand Bazaar was the commercial heart of the old city and
its 4,000 shops are full of treasures including carpets and kilims, silks, jewelry, ceramics, icons, and leather
goods. Wandering through the Grand Bazaar, indulge in some shopping, Ottoman style.
(Afternoon tour also available on Tuesdays)
TOURS
*On Mondays, when Saint Sophia is closed, we visit the Chora Church, famed for its mosaics.
TR-02 OTTOMAN SPLENDOURS (Daily Afternoon Tour)
We begin our tour at Topkapı Palace*, which, from the 15th to the 19th century, was
the principal residence of the Ottoman Sultans. We will visit the fabulous Imperial
Treasury and the Baghdad Kiosk. Topkapı Palace is now a museum and has unrivalled collections of jewelry, including the Spoonmaker’s Diamond, the 3rd largest
in the world. It also possesses numerous Ottoman court costumes and ceramics,
notably including one of the world’s finest collections of Chinese celadon ceramics, many of which were gifts from other rulers. Interestingly, some of the ceramics
have a special glaze that was said to change color in the presence of poison. We
also visit the Imperial Armory, displaying centuries of Ottoman weaponry. But perhaps the loveliest feature of Topkapı Palace are its courtyards with their ancient
trees; it is easy to imagine the sultan strolling here far from the cares of state and
empire. Last but by no means least, we visit the Rüstem Pasha Mosque, completed in 1563. A masterpiece designed by Sinan the Magnificent, it has the most
exquisite and extensive Iznik tile decoration of any mosque in İstanbul. The large
quantities of exquisite Iznik tiles, arranged in a wide variety of beautiful floral and
geometric designs, as well as the spacious central courtyard, make this mosque a
must-see İstanbul attraction.
*On Tuesdays, when Topkapı Palace is closed, this tour is not offered.
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Combining tours TR-01 and TR-02
TR-04 MORNING BOSPHORUS CRUISE (Daily Morning Tour)
We begin with a brief visit to the 17th century Spice Bazaar*, one of İstanbul’s most
colorful, bustling attractions. Next, we travel
the Golden Horn on our way to an unforgettable cruise along the Bosphorus, the majestic strait that runs through İstanbul, linking
Europe and Asia. From our cruise boat, we
view the dramatic sights lining the Bosphorus’
wooded shores: mosques, a bridge that for
a time was the world’s longest and Rumeli Hisar, a massive fortress built by Mehmet the Conqueror in just
three months as he prepared to take İstanbul. Also noteworthy on this tour are the 19th century mansions of the
Ottoman elite and the Sultans’ fanciful gingerbread palaces and hunting lodges.
*On the rare Sundays when the Spice Bazaar is closed, we offer an orientation session.
TR-05 AFTERNOON BOSPHORUS CRUISE (Daily Afternoon Tour)
We cruise the Bosphorus, and from the deck of
our cruise boat, take in the sights and sounds
of this legendary waterway, lined with historic
villages, grandiose waterfront mansions, imposing fortresses, like Anadolu Hisarı, and the
Baroque palaces of the late Ottoman sultans.
After our cruise we travel along the breathtaking Golden Horn to the Spice Bazaar*, a
thrilling riot of colors, sounds and the smells
of exotic spices.
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TR-03 SPECIAL: FULL DAY OLD CITY
*The Spice Bazaar may be visited prior to
the Bosphorus Cruise, depending on the
number of tour participants.
TR-06 ASIA (Daily Afternoon Tour)
We begin by driving across the first Bosphorus
Bridge, which for a time was the world’s longest
suspension bridge, and head for the summit of
Çamlıca Hill, which affords panoramic views of
İstanbul, the Sea of Marmara and the Princes’
Islands. From here a short drive brings us to the
Palace of Beylerbeyi on the shore of the Bosphorus. Perhaps the most elegant of the late
Ottoman palaces, Beylerbeyi boasts six sumptuously furnished reception halls with Bohemian crystal chandeliers and
Sèvres and Chinese vases, including a main salon with an indoor fountain. The sultans’ guests at the palace included
Empress Eugénie of France, Shah Nasruddin of Persia and Grand Duke Nicholas of Russia. (Every day except
Mondays and Thursdays.)
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Combining tours TR-04 and TR-06
The combination of The Morning Bosphorus Cruise and Afternoon Bosphorus Cruise Tours (TR-04 & TR-06)
TR-08 İstanbul BY NIGHT
We visit an exclusive nightclub where we will
enjoy authentic Turkish cuisine, including the
delicious array of appetizers known as meze.
The Turks are justly proud of their food. They
are equally proud of their traditional music,
and after dinner folk musicians from different
regions of Anatolia will perform for us. And
of course no night out in İstanbul would be
complete without belly-dancers. We will be
entertained by some of the city’s finest.
TR-09 THE PRINCES ISLANDS (Daily Full-day)
There is no better way to escape the bustle
of İstanbul for a day than with a visit to the
idyllic Princes’ Islands. Enjoy cool breezes and
charming sights along the way to Büyükada,
the largest Island in the chain. Famous for
their mild climates, lush vegetation, and ornate
Ottoman houses, all the islands are unspoiled
by traffic. Instead of cars there are carriages,
called phaetons, which we will use to tour the island and its beautiful scenery studded with elegant mansions
draped with purple bougainvillea, reminiscent of a more leisured and graceful era. We will enjoy our lunch at the
best of one of the many excellent fresh fish restaurants that line the waterfront, gazing across the Asian shore of
İstanbul, so close – though it feels worlds away.
TR-07 SPECIAL: FULL DAY BOSPHORUS TOUR
TR-10 BURSA & CUMALIKIZIK VILLAGE ((Full-day)
y)
We travel by ferry to Yalova, then by luxury
coach to the city of Bursa*, which dates back
to the 2nd century B.C. The first capital of the
Ottoman Empire, Bursa is one of the greatest
treasure houses of Islamic architecture.
We will first visit the Green Mosque and the
neighbouring Green Mausoleum, both worldrenowned for their superb tile decorations, as
well as the late 14th century Great Mosque and the Koza Han, which for centuries was the center of Bursa’s
flourishing silk trade. As the capital of Turkey’s silk production and importation, Bursa was also the home of
the skilled artisans who produced kaftans, pillows, embroidery and other silk products for the Ottoman palaces
until the 17th century. Afterwards, enjoy a leisurely cable car ride up the slopes of Mount Olympus, in Turkish,
Uludağ, or if the cable car is not running, take in the panoramic views from beneath the more than 1,000 yearold plane tree that gives Çınaraltı its name. Lunch is at one of Bursa’s most highly-regarded traditional Turkish
restaurants, the Darüzziyafe, where we will enjoy the finest authentic Ottoman cuisine in a historic setting,
where Sultan Murat the Second once dined. (This tour is available on Mondays, Thursdays & Saturdays. In the
event the cable car is closed due to weather conditions, Çınaraltı is substituted for Uludağ.)
*A minimum of eight (8) participants are required for this tour.
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TRP-01 TWO-DAYS OF CAPPADOCIA
On our way to Cappadocia, we fly to Ankara or Kayseri and
drive to Üçhisar in the midst of the Cappadocian heartland,
where we will be staying at one of Cappadocia’s unique
boutique Cave Hotels. Cappadocia is a region unlike
any other. Its extraordinary landscape is composed of
soft volcanic stone that wind and rain have sculpted into
fantastic clusters of multicolored spires and pinnacles. What
is more, the softness of the stone encouraged the medieval
Christian inhabitants of the region to carve out of the living
rock literally hundreds of churches, monasteries, towns
and villages. A trip to Cappadocia is a magical experience
you will never forget. On our way to Üçhisar we will stop
off at the magnificent 13th century Selçuk caravanserai
known as Ağzıkarahan, and at one of the region’s famous
‘underground cities’. These go down many storeys into the
earth and are among the marvels of medieval engineering.
Once in Üçhisar, we will visit the great rock-hewn citadel
that looms over the town. On our second day we will visit
the open-air museum of the Göreme Valley, which contains
the region’s highest concentration of painted churches. The
frescoes are among the masterpieces of Byzantine art. A
short distance beyond Göreme lie the villages of Avcılar and
Çavuşin, both of which have many cave-dwellings, and the
dramatic Zelve Valley.
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PRE & POST CONGRESS TOURS INFORMATION
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We catch an early flight to the city of İzmir on the Aegean coast, then drive south to the antique city of
Ephesus. This is one of the world’s richest and most rewarding archaeological sites with impressive ruins
dating from the Classical Greek, Hellenistic, Roman and early Byzantine periods. It is also closely associated
with the beginnings of the Christian faith. In addition to being the city to which the Letters to the Ephesians
were written, St. John and St. Paul both visited the city, and the Virgin Mary herself is said to have spent
her last days nearby. Among the highlights of our tour of Ephesus will be the antique theatre, the famed
Library of Celsus, the brothel, the vast Basilica of St. John, the Mosque of Isabey, the Temple of Artemis (one
of the Seven Wonders of the Ancient World) and a group of recently excavated early Byzantine mansions
that feature incredibly well-preserved floor mosaics and frescoes. At Ephesus, the past comes to life in
a pageantry of architectural and artistic marvels the like of which can be found almost nowhere else in
the world. Finally we will make a short pilgrimage to the House of the Virgin Mary, which stands high on a
mountainside overlooking the Aegean Sea and the Greek island of Samos.
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TRP-02 FULL-DAY TOUR OF EPHESUS
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TRP-03 FULL-DAY TOUR OF TROY AND GALLIPOLI (Upon Request)
Our tour starts with an early departure for Çanakkale, scene of the epic First World War amphibious landing.
After touring Çanakkale, we cross the Hellespont to the battlefields of the Gallipoli peninsula, the crucible
where the national consciousnesses of Australia and New Zealand were forged in steel and blood. Planned
by Winston Churchill, the Gallipoli invasion was the first modern amphibious landing, and one of Turkey’s
greatest military victories of modern times, but it was won at the cost of enormous losses on both sides,
particularly among the heroic ANZAC (Australian and New Zealand Army Corps) troops. They landed
at what is known as Anzac Cove on April 25, 1915, and fought there eight months in a courageous but
doomed struggle. Over 300.000 men from all sides were killed or wounded in the battle’s eight furious
months, making it one of the bloodiest campaigns in history. In the words of Mustafa Kemal Atatürk, Turkish
commander at the battle and founder of the modern Republic, “There is no difference between the Johnnies
and the Mehmets where they lie side by side in this country of ours... Having lost their lives on this land they
have become our sons as well.”
The tours might be organized according to the number of participants.
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PLATINUM SPONSOR
GOLD SPONSOR
BRONZE SPONSORS
S UP P OR T I N G I N S T I T UT I ON S & EX H I BI T ORS
S U PP O RTI NG I NSTI TU TIONS & EXHIB IT OR S
SUPPORTERS
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E X H IB ITI ON FL OOR PL A N
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NO
COMPANY
Sqm
1
2
3
4
12
13
14
15
20-21-22-23
24-25-26-27
Leica
SH 2015
Basel 2016
Celgene
Cell Marque
ATQ / Genova
Cancer Genetics
Abbott
Roche Ventana
3D HISTECH
6 Sqm
Info Desk
Info Desk
9 Sqm
9 Sqm
6 Sqm
6 Sqm
4 Sqm
24 Sqm
24 Sqm
| EAHP - 2014
| 17-22 October 2014
Scientific Secreteriat
Prof. Isinsu Kuzu, MD
University of Ankara, School of Medicine
Department of Pathology, Morphology Building Level 1
06100 Ankara, Turkey
Phone : +90 312 595 81 24 / +90 312 595 80 69
E-mail : scientific@eahp2014.org
Organization Secreteriat
Serenas International Tourism Congress Org. Co.
Turan Güneş Bul. 5. Cad. No: 13 06550 Yildiz, Ankara, Turkey
Phone : +90 312 440 50 11
Fax
: +90 312 441 45 62
URL
: www.serenas.com.tr
E-mail : organizing@eahp2014.org
| 17-22 October 2014
| EAHP - 2014
|
C ORRES P ON DEN C E
C O RRE SPOND E NCE
153
Abate, Francesco 54, 61, 97
Abdelmotagally, Rasha 126
Abdullah, Shahed 84
Abken, Hinrich 60
Acikalin, Arbil 73
Adkins, Elizabeth B. 78
Agostinelli, Claudio 57, 65, 87, 88, 95
Aguilar, Lorena Viramontes 94, 124
Ahlberg, Åsa Jeppson 107
Akarca, Ayse U. 65, 74, 88, 105, 119
Akioka, Toshikazu 80
Akkaya, Bahar 114, 115, 124
Akkaya, Hampar 114, 124
Akkoz, Hayriye Ergin 126
Akoglu, Gulsen 131
Akturk, Seda 108
Aladily, Tariq N. 72
Alaibac, Mauro 84
Aledavood, Amir 127, 128
All, Howayda Sayed Abd El 126
Almeida, Antonio 113
Alnajjar, Khalil 113
Alsancak, Perihan 73
Altay, Hakan 130
Amato, Teresa 61, 98
Ambrosio, Maria Raffaella 61, 92, 97
Anagnostopoulos, Achilles 99, 114
Anagnostopoulos, Ioannis 71
Angeles, Arturo Angeles 124
Angelopoulou, Maria K. 66
Anguita, Eduardo 67
Anila, K.R. 103
Ansell, Stephen M. 53
Antic, Darko 75
Aoun, Patricia 99
Appenzeller, Silke 112
Arai, Satoru 125
Arcaini, Luca 60, 84
Arcila, Maria E. 96
Argüelles, Hugo Álvarez 121
Arra, Mariarosa 60
Asano, Naoko 116
Ascani, Stefano 84
Ashton-Key, Margaret 56
Aster, Jon C. 53
Atalay, Figen 75, 77, 130
Atesoglu, Elif Birtas 75
Attygalle, Ayoma 88
Audry, Georges 110
Aydın, Abdullah 122, 131
Aydın, Funda 115
Aydogdu, Ozge Basaran 125
Aygunes, Duygu 114
B
Baarlen, Joop Van 56
Bağır, Emine Kılıc 73
Bai, Alexandra Papoudou 91
Baia, Maryse 57, 105
Bakan, Ali 131
Balatzenko, Gueorgui 107
Balatzenko, Gueorgui N. 95
Balharek, Tomas 73
Bang, Hae In 76
Barbanis, Sotirios 122
Barbouti, Alexandra 91
Barcena, Carmen 99
Barrans, Sharon 59
Barreira, Rui 113
Basheska, Neli 110
Bastard, Christian 57
Baxter, Richard H.G. 55
Bayık, Mahmut 75
Beato, Margarita Sanchez 67
Becker, Petra S.A. 60
Bedekovics, Judit 71, 72
Bekafigo, Irena Seili 77
Beke, Lívia 72
Bellan, Cristiana 54, 61, 97, 98
Bellas, Carmen 123
Bench, Anthony 56
Benitez, Braulio Martinez 124
Berger, Michael F. 54
Bergman, Christiane Copie 57, 99, 105
Bernd, Heinz Wolfram 87
Bernstein, Bradley E. 62
Berti, Emilio 57, 84
Beverdam, Janetta 56
Bhatt, Ami 53
Bigerna, Barbara 55
Bisig, Bettina 99
Bi, Yingwen 78
Blachnio, Katarzyna 100
Bobadilla, Jose Tovar 94, 124
Boddicker, Rebecca L. 85
Boéchat, Chloé 99
Boer, Jan Paul De 58
Bogdanovic, Andrija 74
Bogusz, Agata M. 55
Bolli, Niccolo 56
Bonfichi, Maurizio 60
Bonis, Carolina De 121
Bont, Eveline De 56
Bonzheim, Irina 68, 90, 93
Boricic, Novica 126
Borisova, Iva 121
Bosch, Ramon 123
Boudjemaa, Sabah 110
Boutsis, Dimitrios 114
Boveri, Emanuela 84
Boz, Adil 115
Bozdag, Zehra 74, 127
Brand, Michiel Van Den 61, 77
Brown, Peter 86
Brugger, Kim 56, 59, 78
Brunnberg, Uta 96
Brynes, Russell K 64
Buechner, Stanislaw A. 96
Bueso-Ramos, Carlos E. 98
Busarla, Satya V. Prasad 92
Bystydzienski, Zbigniew 100
C
Cabecadas, Jose 113
Campo, Elias 63
Carbonell, Felix 67
Carrió, Anna 63
Carvalho, Susana 113
Casey, Laura 119
Castillo, Paola 63
Catherwood, Mark 117
Cazorla, Maite 63
Cazzola, Mario 60
Cerroni, Lorenzo 57
Cetin, Nesibe Kahraman 71
Cetin, Zafer 114
Chamizo, Cristina 89
Chang, Sheng Tsung 103, 106, 113
Chan, John 99
Chatziandreou, Ilenia 66
Chen, Bo Jung 113
Chen, Chih Jung 103
Chen, Jing 78
Chen, Qing Ching 108
Chen, Sonja 109
Chen, Yi Hua 82
Choe, Ji Young 94
Chong, Yosep 118
Chou, Shih Cheng 103
Christensson, Birger 107
Christoforidou, Barbara 122
Chuang, Shih Sung 78, 103, 106, 113
Chung, Stephen S. 54
Chung, Young Rock 54
Chu, Pei Yi 103, 113
Climent, Fina 123
Climent, Jose Angel Martinez 56
Clipson, Alexandra 56, 59
Cloos, Jacqueline 58
Clubwala, Rashna 109
Cogliatti, Sergio 78
Cobanoglu, Bengu 131
Colakkadioglu, Ozge Dilara 127
Collado, Rosa 67
Collie, Angela M. B. 89
Colomer, Dolors 63
Condom, Enric 123
Cordoba, Raul 67
Cork, Kamlai Saiya 53
Coronado, M. Jose 123
Costa, Dolors 63
Cotta, Claudiu V. 115
Coulomb, Aurore 110
Craenmehr, Moniek 117
Croci, Giorgio 84
Croci, Giorgio Alberto 60
Cupic, Dora Fuckar 77
Cvetkovic, Zorica 112
D
Dainese, Linda 110
Dallera, Elena 60
d’Amore, Francesco 129
David, Linch 74
David, Owen 83
David, Schaeffer 83
Daw, Stephen 65
Deangelo, Daniel J. 53
Degroote, Annemarie 83
Delton, Denai R. 72
Demeter, Judit 122
Demiralay, Ebru 75, 130
Dias, Lizalynn 64
Diebold, Jacque 110
Diepstra, Arjan 56
Dieter, Kathrin 93
Dikov, Tihomir 107
Dikov, Tihomir I. 95
Dimopoulou, Maria 66
Dirnhofer, Stephan 81
Divakar, Kiran M. 89
Dogan, Ahmet 96
Dogan, Hayriye Tatli 125, 126
Dogan, Mehmet 126, 131
Doger, Furuzan Kacar 71
Dojcinov, Stefan Dojcin 112
Dorfman, David M. 120
Döring, Claudia 60, 87, 96
Drakos, Elias 106
Drndarevic, Neda Cedomir 112
Dukova, Blagica 110
Du, Ming Qing 56, 59, 78
Dupuis, Jehan 57
Durkin, Lisa 67
Dutilh, Bas 112
A UT H OR I N DEX
A
IN DEX O F A BSTRA CT AU T HORS
E
Ekinci, Deniz 115
Eldaly, Hesham 117
Elgnaoui, Taoufic 105
Elinzano, Heinrich 109
Ellermeier, Chad 109, 127
Ellidokuz, Hulya 104
Epari, Sridhar 86, 120
Epstein, Charles 62
Eray, Mine 96
Erdem, Ramazan 124
Ergin, Melek 73
Ersen, Ayça 104
Esteban, Daniel 63
Esteve, Gloria Pérez 123
Etebari, Maryam 54
| 17-22 October 2014
| EAHP - 2014
|
155
A UT H OR I N DEX
th
156
F
Facchetti, Fabio 57, 87
Falco, Giulia De 54, 61, 92, 97
Falini, Brunangelo 55, 65
Fang, Fang 53
Fasig, Kristina 130
Fazekas, Ferenc 72
Federmann, Birgit 68
Feldman, Andrew L. 85
Feldstein, Julie 54
Feller, Alfred C. 79
Fend, Falko 68, 90, 93
Fend, Leticia Quintanilla 94, 124
Ferretti, Valeria Virginia 60
Fiacacdori, Valeria 60
Firth, Andrew 59
Flevari, Pagona 66
Foe, Jerome Lo Ten 56
Foix, Maria Pane 65
Follows, George 56
Foncillas, Jesús García 89
Fong, Cindy 99
Forshew, Tim 59
Fortini, Elisabetta 55
Frederick, Lori A. 53
Fu, Kai 67
Fuligni, Fabio 54, 57, 97
Furman, Richard R. 53
G
Gaag, Ellen Van Der 56
Gaidano, Gianluca 60
Gainaru, Gabriella 66
Galani, Vassiliki 91
Gambacorta, Marcello 84
Gang, Anne O. 86
Gao, Juehua 82
Garcia, Mar Garcia 77
García, Miguel Teodoro Hernández 121
Gascoyne, Randy 87
Gasljevic, Gorana 76
Gaulard, Philippe 57, 105
Gawronska, Beata Sledz 100
Gazaneo, Sara 92, 97
Gazic, Barbara 76
Gazzola, Anna 93
Gergely, Lajos 122
Gersmann, Ann Kathrin 93
Gert, De Hertogh 83
Gert, Matthijs 83
Geyer, Julia 53
Ghaderi, Mehran 106, 123
Ghafarzadegan, Kamran 127, 128
Gheorghe, Gabriela 74
Ghodke, Kiran 120
Giese, Sabrina 90
Gillissen, Christian 112
Giné, Eva 63, 121
Glaser, Claire 57
Gnandt, Brigitta 120
Go, Heounjeong 125
Gomez, Sagrario 67
Gonlusen, Gulfiliz 73
Gonsalves, Wilson I. 53
Gonzalez, Marcos 67
Goodlad, John 78
Goolsby, Charles L. 82
Goteri, Gaia 84
Gotti, Manuel 60
Goussia, Anna 91
Grada, Zakaria 109
Graf, Mario 96
Granjeaud, Samuel 60
Greipp, Patricia T. 53
Grela, Barbara Pienkowska 100
Grigoropoulos, Nicholas F. 59
Groenen, Patricia 61, 77, 112, 117
Grygalewicz, Beata 100
Gsponer, Joel 81
Guadarrama, Ricardo Aguilar 94
Guenova, Margarita 107
Guenova, Margarita L. 95
Guillermo, Armando López 63
| EAHP - 2014
Gujral, Sumeet 120
Guler, Gulnur 125
Gulmez, Oyku 130
Gundem, Gunes 56
Gunel, Nur Selvi 114
Gungor, Firat 115
Györke, Tamás 122
K
H
Hagiwara, Masahiro 116
Haioun, Corinne 57, 105
Hanafusa, Toshiaki 80
Handy, Beverly 72
Hansmann, Martin Leo 60, 79, 85, 86, 87, 96,
118
Hartmann, Elena 59
Hartmann, Sylvia 85, 86, 87, 96
Havelka, Marija 74
Hay, Ulrich 63
Head, David 130
Hebeda, Konnie 61, 71, 77, 112
Heinrich, Tim 60, 85
Hekimgil, Mine 114
Hermelink, Hans Konrad Muller 110
Hernández, Sonia García 121
Hiddemann, Wolfgang 87
Higashi, Naoyuki 125
Hijmering, Nathalie 58
Hill, Brian T. 67, 89
Hill, Theresa Davies 84
Hiraoka, Nobuya 80
Hirata, Yuji 80
Hirose, Yoshinobu 111
Hitpass, Ludger Klein 55
Hoeller, Sylvia 91
Hoffmann, Karl 90
Høgdall, Estrid 86
Holson, Keegan Barry 64
Hong, Hao 99
Hong, Mineui 89
Hoogendoorn, Mels 56
Horlings, Hugo 82
Horn, Heike 63, 79, 87
Horvath, Emoke 120
Hosone, Masaru 125
Houldsworth, Jane 64
Howard, Matthew T. 53
Hrischev, Vasil G. 95
Hsieh, Pin Pen 106, 113
Hsie, Pin Pen 103
Hsi, Eric D. 67, 89
Hsu, Jeng Dong 103, 106
Huang, Wan Ting 106, 113
Huang, Ying Huang 93
Huang, Yuanxue 59
Huh, Jooryung 94, 125
Hummel, Michael 79, 87
Hunting, Jacco C. 56
Hu, Wenhuo 54
I
Ibarz, Leire Escudero 56, 59, 78
Iltar, Utku 124
Imperi, Elisa 55
Irsai, Gábor 72
Issner, Robbyn 62
Ivancevic, Tijana Dragovic 74, 75
Iwaki, Kazuki 80
J
Jack, Andrew 59
Jacobo, Fernando Perez 94
Jacob, Priya Mary 103
Jaffe, Elaine S. 78, 84, 129
Jain, Shweta 78
Jakobus, Christina 87
Jakovic, Ljubomir 74, 75
Janevska, Vesna 110
Janton, Anna 86
Jayasudha, A.V. 103
Jeon, Yoon Kyoung 94
Jhuang, Jie Yang 103
| 17-22 October 2014
Jong, Daphne De 58
Jonjic, Nives 77
Jovanovic, Maja Perunicic 74, 75
Jovanovik, Rubens 110
Jurisic, Vladimir 126
Juskevicius, Darius 81, 91
Kalpadakis, Christina 66
Kanavaros, Panagiotis 91
Kanellis, George 99, 114
Kang, Chang Suk 118
Kang, So Young 79, 89
Kantarjian, Hagop M. 98
Kanthe, Katha 86
Karadag, Ayse Serap 122
Karakus, Sema 75
Karauzum, Sibel Berker 114
Karmiris, Themistoklis 114
Karube, Kennosuke 63
Kaur, Jasleen 62
Kaygusuz, Gulsah 108
Kazakov, Dmitry V. 96
Keimpema, Martine Van 82
Kempf, Werner 96
Kersten, Marie Jose 82
Kibbelaar, Robby 56
Kim, Annette S. 130
Kim, Eunhee 54
Kim, Hyun Jung 94
Kim, Ji Eun 94
Kim, Tae Jung 118
Kim, Young A. 94
King, Rebecca L. 53
Kinoshita, Tomohiro 116
Kiss, Csongor 72
Kilicarslan, Aydan 125, 126, 131
Kivrak, Hale 108
Klapper, Wolfram 59, 79, 87
Klass, Michael 93
Klausen, Tobias W. 86
Klopcic, Ulrika 76
Kluin, Hanneke C. 56
Kluin, Philip 82
Kluin, Philip M. 56
Kluk, Michael J. 53
Knezevic, Vesna 74
Knoechel, Birgit 53
Knowles, Daniel M. 53
Knudsen, Helle 86
Koc, Eltaf Ayca Ozbal 77
Kokkini, Garifalia 99
Kong, Lilly I. 89
Konoplev, Sergej 98
Konstantinidou, Pavlina 122
Konstantopoulos, Konstantinos 66
Koo, Chiew Loon 103
Korkolopoulou, Penelope 66
Kose, Kenan 108
Kovach, Alexandra E. 55
Kovalchuk, Alexander L. 78
Ko, Young Hyeh 79
Ko, Younh Hyeh 89
Kraan, Willem 82
Kraguljac, Nada 74
Kravtsov, Vladimir D. 2nd 130
Krenács, László 71
Krieken, Han Van 61, 77, 112, 117
Kucukosmanoglu, Aysegul 74
Kuil, Annemieke 83
Kujala, Paula 96
Kuo, Frank 53
Küppers, Ralf 60, 85, 87
Kurtovic, Nada Kraguljac 75
Kuzu, Isinsu 108
Kwiecinska, Anna 106, 123
L
Laar, Lisette Van De 117
Laginestra, Antonella 57
Laginestra, Maria Antonella 54, 97
Lai, Raymond 67
Lakhwani, Sunil 121
M
Ma, Charles 64
Maclennan, Kennan 110
Maffi, Aldo 60, 84
Magnano, Laura 121
Magunacelaya, Nerea Martínez 89
Maldonado, Carmen Lome 94, 124
Malek, Sami N. 53
Mali, Kinga Sikorska 100
Malik, Muhammad Adnan 108
Manilich, Elena A. 89
Mannucci, Roberta 55
Mannu, Claudia 93
Manso, Rebeca 89
Marafioti, Teresa 57, 65, 88, 95, 105, 119
Marass, Francesco 59
Marcinek, Juraj 73
Marco, José Antonio Garcia 67
Marijke, Spaepen 83
Martelli, Maria Paola 55
Martín, Alejandro Martín 121
Martínez, Daniel 63
Martinez, Leticia Quintanilla 68, 90, 93
Martinez, Nerea 67
Martin, Howard 59
Martinovic, Tamara Ivan 112
Martinovic, Vesna Mihajlo Cemerikic 112
Martin, Paloma 123
Mason, Emily F. 120
Masszi, Tamás 122
Masuda, Yuki 80
Mathijssen, Janneke 77
Matolcsy, Andras 123
Medeiros, Jeffrey 98
Méhes, Gábor 71, 72
Melle, Federica 57
Melnick, Ari M. 53, 54
Memar, Bahram 127, 128
Mendes, Larissa Sena Teixeira 88
Menon, Hari 86, 120
Menter, Thomas 91
Meteoğlu, Ibrahim 71
Meyerson, Matthew 53
Michova, Antoaneta S. 95
N
Naeim, Faramarz 115
Nafeeh, Dalia Ahmed 126
Nair, Rekha A. 103
Nair, Sreejith 103
Naito, Zenya 125
Nakamura, Shigeo 116
Nakashima, Megan O. 67
Nakayama, Shoko 80, 111
Nasiri, Mohammad Reza Ghavam 127, 128
Natwani, Bharat 110
Neme, Yvette 124
Neto, Lara 113
Newrzela, Sebastian 60, 85, 86, 87, 118
Nicolae, Alina 78, 84
Nicola, Marta 60, 84
Nicoletti, Ildo 55
Nielsen, Signe L. 86
Nielsen, Signe Ledou 129
Nieto, Luis Hernández 121
Nishiwaki, Uta 80
Nissen, Loes 112
Noessel, Max M. Van 56
Nolling, Jork 89
Noorduyn, Arnold 82
Nørgaard, Peter 86, 129
Pangalis, Gerasimos A. 66
Panganiban, Jeff 93
Pan, Li 53
Pan, Shien Tung 103
Panzacchi, Riccardo 88, 95
Papadaki, Theodora 99, 114
Papanastasiou, Konstantinos 99
Papanikolaou, Asimina 99
Park, Christopher Y. 54
Park, Jae H. 54
Park, Rojin 76
Park, Sanghui 111
Patakiouta, Frideriki 122
Patsouris, Efstratios 66, 88, 106
Paulli, Marco 60, 84
Paydas, Semra 73
Pecos, Patricia 121
Pedersen, Mette Ø. 86
Pedersen, Michael 86, 129
Peeters, Christiane De Wolf 87
Pektas, Gokhan 71
Peterson, Loann C. 82
Petkovic, Jelena 126
Petrusevska, Gordana 110
Pettirossi, Valentina 55
Phan, Ryan 115
Piccaluga, Pier Paolo 54, 57, 61, 87, 93, 97
Picquenot, Jean Michel 57
Pierce, Sherry A. 98
Pileri, Stefano 61, 87, 95
Pileri, Stefano A. 65, 88
Pileri, Stefano Aldo 54, 57, 93, 97
Pimpinelli, Nicola 84
Pinilla, Socorro María Rodríguez 89
Pinyol, Magda 63
Piris, Miguel Ángel 67, 89
Pittaluga, Stefania 84, 129
Plank, Lukas 73
Plata, Eleni 66
Pomplun, Sabine 74
Ponz, Olga Balagué 58
Ponzoni, Maurilio 87
Pouliou, Evdoxia 99, 114
Poulsen, Tim S. 129
Pozdnyakova, Olga 120
Prem, Sruthi 103
Prevodnik, Veronika Kloboves 76
Priebe, Valdemar 107
Pucciarini, Alessandra 55
A UT H OR I N DEX
Miedema, Iris 58
Mihailovic, Dragan Slavoljub 106, 124
Mihaljevic, Biljana 75
Mijovic, Zaklina Zarko 106, 124
Milionis, Vasilis 66
Millan, Isabel 123
Miller, Jeffrey 93
Milosevic, Violeta 75
Minotakis, Dimitrios 122
Miyoshi, Takuji 80
Mohtashami, Samira 128
Möller, Peter 79, 87, 90
Molnár, Dávid 122
Moody, Sarah 56, 59, 78
Mora, Roberto Hernández 94, 124
Morello, Lucia 60
Moreno, Santiago Montes 98
Morice, William G. 53
Morscio, Julie 91
Morse, Herbert C. 78
Mortland, Leslie J. 74
Moschogiannis, Maria 66
Mourmouras, Vasileios 92
Mujdaba, Metin Timur 109
Mundo, Lucia 92, 97
Muniesa, Cristina 123
Lakiotaki, Eleftheria 66
Lambein, Kathleen 83
Lam, King 56
Landi, Sylvaine Just 60
Langerak, Ton 56
Lapadat, Razvan 74
Larue, Marie Hélène Delfau 57, 105
Lautitzen, Anne F. 86
Lazzi, Stefano 92, 98
Lee, Eun Jung 118
Lee, Seung Eun 79
Lehtinen, Tuula 96
Leich, Ellen 87
Le, Long P. 55
Lemale, Julie 110
Lennerz, Jochen K. 90
Leoncini, Lorenzo 54, 61, 92, 97, 98
Le, Suong 60
Leval, Laurence de 56, 99, 105
Leverger, Guy 110
Levidou, Georgia 66, 88, 95
Li, Betty 120
Lies, Knippenberg 83
Li, Hongxiang 78
Linch, David 65
Lin, Jeffrey J. 89
Lin, Pei 72
Liu, Hongxiang 56, 117
Liu, Yi Fang 53
Li, Xianqian 67
Li, Xiaodong 115
Loeffler, Markus 79
Lord, Martin 107
Louie, Carrie 99
Louis, Libbrecht 83
Lozano, Carol 67
Lucin, Ksenija 77
Lucioni, Marco 60, 84
Q
Quaglino, Pietro 84
R
O
O’Brien, Susan 98
Olgun, Aybuke 104
Olive, Daniel 60
Onida, Francesco 84
Orazi, Attilio 53, 74
Ortiz, Catalina Amador 82
Osada, Shin-ichi 125
Oscier, David 56
Ott, German 59, 63, 79, 87
Ott, M. Michaela 63
Oud, Monique 82
Ouillette, Peter 53
Ozcan, Mehmet Ali 104
Ozcan, Mualla 114, 124
Ozkal, Sermin 104
Ozkanlı, Seyma 131, 122
Ozsan, Nazan 114
Ozturk, Erman 122, 131
Ö
Österborg, Anders 123
P
Paik, Jin Ho 94
Pals, Steven 82
Pals, Steven T. 83
Panayiotidis, Panayiotis 66
Raaij, Annemiek Kastner Van 112
Rabadan, Raul 54, 57, 61, 97
Raffeld, Mark 78
Ramil, Elvira 123
Ramponi, Antonio 60, 84
Ramsay, Alan 65
Rassidakis, George Z. 106, 123
Rattotti, Sara 60, 84
Raya, José María 121
Raziee, Hamid Reza 128
Rengstl, Benjamin 60, 85, 86, 87, 118
Rey, Françoise Gondois 60
Riboni, Roberta 60, 84
Rich, Amy 98
Rieger, Michael A. 85
Righi, Simona 88, 95
Rijn, Marieke Van 117
Rijntjes, Jos 112, 117
Rincon, Julia Gonzalez 67
Rintels, Peter 109
Rizvi, Hasan 65, 119
Robles, Eloy F. 56
Robson, Alistair 78
Rocca, Bruno Jim 92
Rodriguez, Antonia 67
Rojo, Federico 89
Rooij, Martin F. De 83
Roopenian, Derry 78
Rosati, Stefano 56
| 17-22 October 2014
| EAHP - 2014
|
157
A UT H OR I N DEX
th
158
Rosenfeld, Nitzan 59
Rosen, Neal 54
Rosenwald, Andreas 59, 63, 79, 87
Rosini, Francesca 95
Rossi, Davide 60
Rossi, Maura 54
Roussou, Paraskevi 99
Royo, Cristina 63
Rozman, María 121
Rózsa, Tímea 72
Rudelius, Martina 59
Rufle, Alexander 81
Ruiz, Sory J. 115
Russo, Guido 55
Rüther, Nele 90
Ryan, Russell J.H. 62
Rygier, Jolanta 100
Rymkiewicz, Grzegorz 100
Stefanaki, Kalliopi 91
Stefanoudaki, Ekaterini 99
Steidl, Christian 87
Stein, Harald 79, 87
Steinhilber, Julia 93
Stevens, Wendy 117
Su, Jing 59
Sukumaran, Renu 103
Sungu, Nuran 126
Süsskind, Daniela 90
Szablewski, Vanessa 57, 105
Szepe, Peter 73
S
Tabanelli, Valentina 57
Tabrizi, Fatemeh Varshoee 127
Takayama, Ayami 80
Talamo, Anna 99
Tallman, Martin S. 54
Tam, Wayne 53, 74
Tasidou, Anna 114
Tatic, Svetislav 74
Tecimer, Tulay 131
Telatar, Milhan 99
Teresa, Marafioti 74
Terre, Christine 57
Terzic, Tatjana 74, 126
Thalheimer, Frederic B. 85
Thomas, Tousseyn 83
Tiacci, Enrico 55
Tímár, Botond 122
Timuragaoglu, Aysen 114
Tinguely, Marianne 96
Todorovska, Magdalena Bogdanovska 110
Tol, Esther Schilder 82
Tomasini, Carlo 84
Tombuloglu, Murat 114
Topalov, Yavor 107
Tops, Bastiaan 112
Toptas, Tayfur 115
Tousseyn, Thomas 87, 91
Tower, Richard L. 74
Trajkova, Sanja 110
Treaba, Diana Olguta 109, 127
Treon, Steven P. 83
Trougouboff, Philippe 116
Tsionos, Konstantinos 114
Tsou, Mei Hua 106
Tsuji, Motomu 80, 111
Turcu, Mihai Lazar 120
Tzankov, Alexandar 71, 81, 91
Sabattini, Elena 88, 95
Sabbioni, Simone 88
Sacchetti, Carlo Sagramoso 95
Sachanas, Sotirios 66
Sadowska, Justyna 96
Saetta, Angelica A. 66
Saft, Leonie 106, 123
Sagaert, Xavier 87
Sakai, Tomomi 78
Salazar, María José Rodríguez 121
Salim, Ozan 115, 124
Sander, Birgitta 107
Santi, Alessia 55
Santos, Taida Martín 121
Santucci, Marco 84
Sapienza, Maria Rosaria 57
Sara, Vanderborght 83
Sarı, Ibrahim 74, 127
Saydam, Guray 114
Scheepstra, Cornelis 82
Schiavoni, Gianluca 55
Schindler, Natalie 68
Schmid, Frederike 60, 85, 86, 118
Schmidt, Janine 68
Schmidt, Karl Ulrich Bartz 90
Schmitz, Norbert 79
Schroeder, Timm 85
Schuuring, Ed 56
Scott, Mike 56
Seidl, Christian 60
Seldenrijk, Kees 56
Sengar, Manju 86
Sepsa, Athanasia 66
Serrano, Sergio 77
Servitje, Octavi 123
Seymour, Erlene 53
Shahidsales, Soodabeh 128
Shaknovich, Rita 53
Sharifian, Maryam Maryam 115
Sharma, Vandana 99
Shedden, Kerby 53
Shende, Vishvesh 119
Shende, Vishvesh H. 65
Shet, Tanuja 86, 120
Shin, Dong Mi 78
Shin, Soojin 94
Siddiqi, Imran N. 64
Siebert, Reiner 87
Sievers, Cem 62
Silva, Maria 113
Silveira, Margarida 113
Singh, Zeba 108
Smith, Mitchell R. 89
Sobieszek, Monika Prochorec 100
Sohani, Aliyah R. 55
Somja, Joan 65, 105
Son, Eun Mi 125
Soria, Beatriz 121
Spaargaren, Marcel 82, 83
Sretenovic, Snezana 126
Staiger, Annette M. 63, 79
Stamatopoulos, Konstantinos 99, 114
Stavroyanni, Niki 99, 114
| EAHP - 2014
Ş
Sahin, Nurhan 74, 127
Sahin, Yasemin 108
T
U
Undar, Levent 115, 124
Unterhalt, Michael 87
Uysal, Ayse 114
V
Valera, Alexandra 63
Valkovic, Toni 77
Vardarlı, Aslı Tetik 114
Varela, Mar 123
Varettoni, Marzia 60
Vassilakopoulos, Theodoros 66
Vassiliou, George 56
Velden, Walter Van Der 117
Velikonja, Mojca 76
Venkatraman, Lakshmi 117
Vera, Jorge Garcia 94, 124
Viardot, Andreas 90
Vicente, Yolanda 123
Villaespesa, Miguel Piris 67
Visser, Lydia 56
Viswanatha, David S. 53
Vöhringer, Matthias 63
Vornanen, Martine 96
Vural, Filiz 114
| 17-22 October 2014
W
Wahab, Omar Abdel 54
Waldmann, Thomas A. 78
Wang, Hongsheng 78
Wang, Ming 56, 59, 78
Wang, Xiaohong I. 72
Ward, Jerrold M. 78
Warner, Kathrin 60, 85, 86
Wartenberg, Martin 79, 87
Wasik, Agata M. 107
Watkins, A. James 56
Wei, Chih Hsin 103
Weisenburger, Dennis 99, 110
Weiser, Christian 60, 85, 86
Weng, Shih Feng 103, 106, 113
Whitton, Holly 62
Wilton, James 65, 119
Wit, Peter De 56
Wlodarska, Iwona 91
Wolniak, Kristy L. 82
Wooler, Gitte 129
Woroniecka, Renata 100
Worrillow, Lisa 59
Wotherspoon, Andrew 56, 88
Wunderlich, Hilka Rauert 59
X
Xagoraris, Ioanna 106, 123
Xavier, Sagaert 83
Xerri, Luc 60
Xirou, Persefoni 122
Xochelli, Aliki 99
Xue, Xuemin 59, 78
Y
Yang, Sheau Fang 106
Yao, Jinjuan 96
Yavasoglu, Irfan 71
Yavuz, Sibel Orhun 131
Yildiz, Akin 115
Yildiz, Semsi 75, 77, 130
Yokote, Taiji 80, 111
Yucel, Orhan Kemal 115, 124
Yung, Evan 115
Yun, Ji Yun 94
Z
Zairis, Sakellarios 54
Zanden, Adri Van Der 56
Zemheri, Ebru 122, 131
Zenginkinet, Tulay 122, 131
Zeng, Naiyan 59, 78
Zettel, Andreas 96
Zhang, Xiaohong Mary 81
Zhao, Xiangrong 129
Zhao, Xiaoxian 67
Ziepert, Marita 79
Zimmermann, Dieter R. 96
Zweegman, Sonja 58
18th Meeting of the European Association
for Haematopathology
EAHP2016
© graberfotografie - Fotolia.com
BASEL · SWITZERLAND | SEPTEMBER 3–8
Þ www.eahp2016.com
Local Organizer
Stephan Dirnhofer, MD
Universitätsspital Basel
Department of Patholgy
Basel, Switzerland
Congress Venue
Congress Center Basel
MCH Messe Basel
Messeplatz 21
4058 Basel, Switzerland
Legal Organizer (PCO)
MCI Deutschland GmbH
MCI – Berlin Office
Markgrafenstr. 56, 10117 Berlin, Germany
Phone: +49 (0)30 20 45 90
E-mail: eahp@mci-group.com
VENTANA hematopathology diagnostics
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The VENTANA hematopathology menu:
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Includes important assays such as c-myc, SOX-11, CD13, and TdT to aid in the diagnosis of common
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Consists of >65 ready-to-use, highly sensitive and specific rabbit and mouse monoclonal assays and
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To learn more, stop by the VENTANA booth or visit www.ventana.com.
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© 2013, 2014 Ventana Medical Systems, Inc.
VENTANA and the VENTANA logo are trademarks of Roche.
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