Cell-mediated immune responses generated after DNA delivered by

Cell-mediated immune responses generated
after DNA delivered by either Biojector or
Electroporation and boosted with a
heterologous insert recombinant poxvirus
September 13, 2011
Jeffrey R. Currier,
C
PhD
Military HIV R
Milit
Research
hP
Program
Division of Retrovirology, WRAIR
Background
• RV262: Phase I study of the safety and immunogenicity of
g) administered by
y either
PENNVAX™-G DNA ((Env and Gag)
Biojector® 2000 or by CELLECTRA® Intramuscular
Electroporation device followed by administration of MVACMDR (HIV-1
(HIV 1 CM235 env/CM240
en /CM240 gag/pol boost in healthy,
health
HIV uninfected adults
• Part A:
A Open-Label
Open Label safet
safety study
st d conducted
cond cted at the MHRP in
Rockville, MD in 12 healthy, HIV uninfected adults
• Part B:
B Randomized
Randomi ed placebo
placebo-controlled
controlled st
study
d opening in
Kenya, Tanzania and Uganda (Oct’ 2011) with an n=80, HIV
uninfected adults
PENNVAX™-G DNA (Prime)
• Consists of 4 DNA plasmids:

3 plasmids each e
expressing
pressing an HIV Env
En consensus
consens s modified
gp140 proteins for subtypes A, C and D respectively

1 plasmid encoding a Gag consensus
consens s protein deri
derived
ed from
subtypes A, B, C and D
• Administered in 4 mg doses,
doses 1 ml volume (days 0
0, 28)
28):

2mg of the Gag expressing plasmid

0.7 mg of each Env expressing plasmid
• Two delivery
y methods using
g either:

BIOJECTOR® device: needle-free injection intra-muscularly

CELLECTRA® device:
de ice in vivo
i o intra-muscular
intra m sc lar electroporation
MVA-CMDR (Boost)
• Live, non-replicating, double recombinant, MVA vector

Expressing CRF01_AE Env (gp140) with a truncated
cytoplasmic
t l
i tail
t il

Expressing CRF01_AE Gag/Pol native open-reading frame
(i t
(integrase
d l t d and
deleted
d reverse ttranscriptase
i t
functionally
f
ti
ll
inactivated)

108 pfu
f administered
d i i t d iintra-muscularly
t
l l on study
t d days
d
84,
84 168

Proven safety and immunogenicity as a stand-alone product
RV262 DNA/MVA Study Schedule
PENNVAX™- G DNA
((EP or BioJect))
6-month vaccination
schedule
MVA-CMDR
0
1
3
6
12
27
(time in months)
V2
V7
V10
V13
V15
Immune monitoring schedule
IFN- Elispot and Multi-functional ICS
Pre-vaccination and 2 weeks post DNA and
each MVA vaccination
RV262 Preliminary Safety – Part A
• First-in-man safety evaluation of PENNVAX™-G:
 Consensus Env A, C, D, and CG4 plasmids
• Testing in vivo electroporation versus Biojector for DNA delivery
• Total n=13 subjects
j
enrolled, 2 studyy unrelated withdrawals
• No related SAEs, no pregnancies, no HIV infections
• To date, DNA prime and boosting with MVA has been well
tolerated
RV262 Cellular Immunogenicity (CMI)
OBJECTIVES
• Compare the relative immunogenicity of electroporation versus Biojector
delivered DNA followed by an MVA boost
• Use a standard IFN- Elispot assay and prime/boost matched peptides
y
using
g flow cytometry
y
y for function and p
phenotype
yp ((14 color,,
• Real-time analysis
16 parameter assay)
 Functions: IFN-,
, IL-2,, MIP-1,
, TNF-,, and CD107
 Phenotyping: Naïve, Central memory, Effector memory, Effector cells
 ICS performed only with boost (MVA-CMDR insert) matched peptides
• PBMC were tested Pre-vaccination,, 2 weeks post
p
2x DNA prime
p
and 2 weeks
posts each MVA boost
HIV-specific Response Rate by IFN- Elispot
HIV Env-specific Response by IFN- Elispot
193
146
((62-555)) ((12-367))
MVA-CMDR
ENV-PP
(CM235)
76
98
(72-342) (15-215)
PENNVAX-G
ENV C PP
ENV-C-PP
12
32
((3-65)) ((23-72))
Point Prevalence for T Cell Responses (ICS)
• Positive response >0.05% of all antigen-specific T cells (and 3x BG)
• Functional assessment has the caveat of boost (MVA) matched peptides
HIV antigen-specific CD4 T cells (any function)
MVA-CMDR
ENV-PP
(CM235)
MVA-CMDR
GAG-PP
(CM240)
HIV antigen-specific CD8 T cells (any function)
MVA-CMDR
ENV-PP
(CM235)
MVA-CMDR
GAG-PP
(CM240)
HIV-specific T Cell Responses by Function
Gating Strategy: Effector/EM/CM Discrimination
CD8 T Cells
EFFECTOR MEMORY
CD45RO+/CD45RA+/
CCR7-/CD28+/-
CENTRAL MEMORY
CD45RO+/CD45RACCR7+/CD28+
EFFECTOR T CELL
CD45RA
C
5 +/C
/CD45RO
5 OCCR7-/CD28+/-
NAIVE T CELL
CD45RA+/CD45ROCCR7+/CD28+
HIV-specific T Cell Responses by Phenotype
Effector memory CD4+ T cells are generated
CD4+
T cells
EFFECTOR MEMORY
CD45RO+/CD45RACCR7-/CD28+/-
CENTRAL MEMORY
CD45RO+/CD45RACCR7+/CD28+
EFFECTOR T CELL
CD45RA+/CD45ROCCR7-/CD28+/-
NAIVE T CELL
CD45RA+/CD45ROCCR7+/CD28+
Eff / Effector memory CD8+ T cells are generated
CD8+
T cells
EFFECTOR MEMORY
CD45RO+/CD45RACCR7-/CD28+/-
CENTRAL MEMORY
CD45RO+/CD45RACCR7+/CD28+
EFFECTOR T CELL
CD45RA+/CD45ROCCR7-/CD28+/-
NAIVE T CELL
CD45RA+/CD45ROCCR7+/CD28+
Preliminary Conclusions
• Few responses detected post-DNA using either MVA-matched peptides
(ICS) or DNA-matched peptides (Elispot)
• After the 1st MVA boost:
 11/11 subjects and 5/11 subjects respectively have Env-specific CD4
and
d CD8 T cellll responses iin th
the 0.05
0 05 – 0.57%
0 57% range
 Gag responses seen in 6/11 and 2/11 subjects for CD4 and CD8 T
cells
ll respectively
ti l after
ft th
the 1st MVA vaccination
i ti
• CD8 responses are predominantly EM after the MVA boosts
• CD4 responses are a balance of EM and CM (2:1) after the MVA boosts
• Elispot
Eli
t responses iin di
directt agreementt with
ith th
the ICS d
data
t
• Diminution of response rate and magnitude after 2nd MVA vaccination
Acknowledgements
MHRP Rockville
Mary Marovich, Protocol Chair/PI
Tina Tong, Manager, Bioproduction
Tyesha Jackson, GLP/QC Lab
Silvia Ratto-Kim, Co-Investigator
Merlin Robb
Jerome Kim
Nelson Michael
AFRIMS, Bangkok
Mark de Souza, Laboratory Director
Viseth Ngauy, Protocol co
co-PI
PI
MHRP Cellular Immunology Laboratory
Doris Thelian
Erick Herrera
Vicky Lo
Commercial Partners
N. Sardesai, Innovio
A. Khan, Innovio
J. Yan, Innovio
M. Bagarazzi, Innovio
MHRP Vaccine Clinical Research Clinic
Julie Ake
Cheun Yen Lau
Lei Zheng
Laboratory of Viral Diseases, NIAID
Patricia Earl
Leigh Anne Eller
Bernard Moss
University of Pennsylvania
D Weiner
D.
J. Boyer
NIH Division of AIDS and DoD: Funding,
Funding Support,
Support Sponsorship,
Sponsorship Oversight
Supplemental Slides
Vector-specific CD8 T cell Response Boosting