Rôle des cellules NK dans l`immunité de la peau et des muqueuses

22/02/2011
Rôle des cellules NK dans
l'immunité de la peau et des
muqueuses
Thierry Walzer
thierry.walzer@inserm.fr
INSERM U851 Lyon
Innate Immunity
Effector
Functions
« Pathogens »
Initiation of
Ag-specific
response
Effector
Functions
Adaptive Immunity
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Lymphocyte development
CLP
Pro-B
Pre-T
NKp
T CD4
B cells
NK
NKT
γδ T
T CD8
Treg
TH1 TH2 TH17
Third lineage of lymphocytes
Cytotoxic lymphocytes
NK cell phenotypes
• Humans: CD3-CD56(N-CAM)+
• C57BL/6 mice: CD3-NK1.1+
• Other mice: CD3-DX5(CD49b)+
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NK Cells - Where do they live?
• ~5-20% peripheral blood lymphocytes
(human, 5% in mice)
• ~5% lymphocytes in spleen
• Abundant in liver and lung
• Low frequency in thymus, bone marrow,
uninfected lymph nodes and lymphatics
• >90% of lymphocytes in decidual tissue
NK cells: effector functions
Perforin-dependent cytotoxicity
IFN-γ secretion
chemokines
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NK cells: regulation of other cell
types
Fonctional duality: effector or regulatory cell
Vivier et al, Nature Immunology 2008
Strategies of NK cell recognition
Normal cell
MHC class I
Activating ligands
NO ACTIVATION
Activating receptors
+
-
Inhibitory KIR/Ly49
NK cell
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Strategies of NK cell recognition
« stressed »
cell
MHC class I
Activating ligands
ACTIVATION
Inhibitory KIR/Ly49
-
+
Activating receptors
NK cell
Vivier and Malissen, Nature Immunology 200
Strategies of NK cell recognition
Toll-like Receptors
NOD molecules +
Ly49H
NKG2D
+
KIR, Ly49
CD94/NKG2
NKRP-1
-
Infectious non-self
Stress-induced self
(tumors)
Missing self
(tumors)
Microbial molecules
MICA/B, ULBP/RAET
Rae1, H60, MULT1
Classical MHC class I
Non-classical MHC class I
Clrg/OCIL
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NK cells: role in immunity
Main effector
Herpesviruses
NK-sensitive tumors
Stem cell transplantations
Regulatory
(protective)
redundant
effector
Organ transplantation
Autoimmunity
inflammation
allergies
Other viruses
Intracellular bacteria
Intracellular parasites
NK cells: role in immunity
Main effector
Herpesviruses
NK-sensitive tumors
Stem cell transplantations
Regulatory
(protective)
redundant
effector
Organ transplantation
Autoimmunity
inflammation
allergies
Other viruses
Intracellular bacteria
Intracellular parasites
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NK cell role in anti-viral
responses-evidences
• Concept of immunology « in natura » (JL Casanova) :
Mice depleted of NK cells are more sensitive to
challenge by many viruses, intracellular bacteria and
parasites. But this is experimental.
– Æ Let see what human patients specifically deficient in NK cells
suffer from
• Æreccurent infections by herpes viruses (CMV, HSV, varicella, EBV)
Problem : very few patients (6?)
• ÆIntricate relationship between NK cells and herpesviruses
Cas des herpes virus
CMV
HSV
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Evolutionary success:
• High transmissibility
• Low pathogenicity
• Persist in a latent state
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MCMV-infected cell
NK cell
L’expansion des NK Ly49H+ ressemble à une expansion clonale de LT en réponse
à un antigène. Des cellules NK survivent à cette expansion clonale et ont des
fonctions améliorées (NK « memoire »)
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Case of CMV
But CMV downregulates NKG2D ligands
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Conclusion: Co-adaptation of CMV and the
host immune system through evolution
Cas de HSV
• Infection le plus souvent localisée au niveau des
muqueuses, récurrente mais bénigne. Chez certains
patients, l’infection est plus sévère et se propage au
CNS
• Déplétion NK chez la souris sensibilise à l’infection. La
migration de NK/CD8 au niveau de la lésion est cruciale.
Pas de récepteurs connus.
• Au niveau génétique des mutations dans TLR3 et
UNC93B1 prédisposent à l’infection.
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NK cells: role in immunity
Main effector
Herpesviruses
NK-sensitive tumors
Stem cell transplantations
Regulatory
(protective)
redundant
effector
Organ transplantation
Autoimmunity
inflammation
allergies
Other viruses
Intracellular bacteria
Intracellular parasites
NK cells / bacteria : textbooks
• NK cells in bacterial infection
– TLR signaling of myeloid cells induces IL-12
– IL-12 induces NK cells to make IFNγ
– IFNγ augments macrophage function
IFNγ
NK cell
macrophage
IL12/18
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L’IL12 et l’IFNγ sont crucials pour la protection contre les mycobactéries
mais pas essentiels pour la protection contre d’autres pathogènes
Msmd = mendelian susceptibility to mycobacteria
From JL Casanova
IFNγ producing cells: NK paralogs
IFNγ
•
•
•
•
•
•
NK cells
CD1d restricted NKT cells
Other NKT cells
Gamma/delta T cells
Memory CD8 T cells
TH1 CD4 T cells
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NK cells: role in immunity
Main effector
Herpesviruses
NK-sensitive tumors
Stem cell transplantations
Regulatory
(protective)
redundant
effector
Organ transplantation
Autoimmunity
inflammation
allergies
Other viruses
Intracellular bacteria
Intracellular parasites
NK cells in allergies / autoimmunity :
descriptive, correlative data
• Lung (asthma)
– NK cells are rather protective
– NK cell deficiencies predispose to viralinduced TH2-type allergic inflammation
– But: NK cells may (?) differentiate into Th2
cytokine producing cells and contribute to the
inflammation
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NK cells in allergies / autoimmunity :
• Skin :
– Atopic dermatitis
– Psoriasis
– Alopecia aerata
– Pemphigus vulgaris
– Others
Problems to detect NK cells in situ
(No specific marker)
Identification of NK cells
–
–
–
–
–
Humans: CD3-CD56(N-CAM)+
C57BL/6 mice: CD3-NK1.1+
Other mice: CD3-DX5(CD49b)+
Rats: CD3-NKR-P1+
Non-human primates: CD3-CD16+?
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Collection of microarray data for most cell types
in both mouse and human
mouse NK cell signature
human NK cell signature
mammalian NK cell signature
The human NK cell signature
Cytokines / chemokines / secreted proteins
IFNG
interferon, gamma
CCL4
chemokine (C-C motif) ligand 4
XCL1
chemokine (C motif) ligand 1
XCL2
chemokine (C motif) ligand 2
SPON2
spondin 2, extracellular matrix protein
IL15 response
IL2RB
interleukin 2 receptor, beta
Cytotoxic granules
CST7
cystatin F (leukocystatin)
GNLY
granulysin
GZMH
granzyme H (cathepsin G-like 2, protein h-CCPX)
PRF1
perforin 1 (pore forming protein)
NK receptors
KIR2DL1
killer cell immunoglobulin-like receptor, two domains, long 1
KIR2DL3
killer cell immunoglobulin-like receptor, two domains, long 3
KIR2DL4
killer cell immunoglobulin-like receptor, two domains, long 4
KIR2DS1
killer cell immunoglobulin-like receptor, two domains, short 1
KIR2DS2
killer cell immunoglobulin-like receptor, two domains, short 2
KIR2DS5
killer cell immunoglobulin-like receptor, two domains, short 5
KIR3DL1
killer cell immunoglobulin-like receptor, three domains, long 1
KIR3DL2
killer cell immunoglobulin-like receptor, three domains, long 2
KLRC3
killer cell lectin-like receptor subfamily C, member 3 (NKG2E)
KLRD1
killer cell lectin-like receptor subfamily D, member 1 (CD94)
KLRF1
killer cell lectin-like receptor subfamily F, member 1 (NKp80)
LAIR2
leukocyte-associated Ig-like receptor 2
CD160
CD160 antigen
CD244
CD244 natural killer cell receptor 2B4
CD300A
CD300A antigen
NCR1
natural cytotoxicity triggering receptor 1 (NKp46)
NCR3
natural cytotoxicity triggering receptor 3 (NKp30)
NKG7
natural killer cell group 7 sequence (GMP-17)
Other transmembrane proteins
EDG8
endothelial differentiation, sphingolipid G-protein-coupled receptor, 8
FASLG
Fas ligand (TNF superfamily, member 6)
CX3CR1
chemokine (C-X3-C motif) receptor 1
GPR65
G protein-coupled receptor 65
LGR6
leucine-rich repeat-containing G protein-coupled receptor 6
LIM2
lens intrinsic membrane protein 2, 19kDa
PTGDR
prostaglandin D2 receptor (DP)
PTGER2
prostaglandin E receptor 2 (subtype EP2), 53kDa
SIGLECP3
sialic acid binding Ig-like lectin, pseudogene 3
IL18RAP
interleukin 18 receptor accessory protein
HAVCR2
hepatitis A virus cellular receptor 2
Adapter proteins
EAT2
SH2 domain-containing molecule EAT2
GNG2
guanine nucleotide binding protein (G protein), gamma 2
SH2D2A
SH2 domain protein 2A
Signaling
MAPK7
PRKCH
PTPN12
RASA3
RASSF1
mitogen-activated protein kinase 7
protein kinase C, eta
protein tyrosine phosphatase, non-receptor type 12
RAS p21 protein activator 3
Ras association (RalGDS/AF-6) domain family 1
Transcription factors
RUNX3
runt-related transcription factor 3
TBX21
T-box 21
ZNF595
zinc finger protein 595
Cytoskeleton
SYNE1
spectrin repeat containing, nuclear envelope 1
MYOM2
myomesin (M-protein) 2, 165kDa
Others / unkown
ABI3
ABI gene family, member 3
ASB1
ankyrin repeat and SOCS box-containing 1
AXIN1
axin 1
C21orf106
chromosome 21 open reading frame 106
C9orf81
chromosome 9 open reading frame 81
CXorf26
chromosome X open reading frame 26
DOCK11
dedicator of cytokinesis 11
FLJ14213
hypothetical protein FLJ14213
FLJ16686
FLJ16686 protein
FLJ20699
hypothetical protein FLJ20699
FLJ21415
hypothetical protein FLJ21415
FLJ30655
hypothetical protein FLJ30655
FLJ35036
hypothetical protein FLJ35036
FLJ45909
FLJ45909 protein
KIAA0493
KIAA0493 protein
LOC343413 IFGP6
LOC54103
hypothetical protein LOC54103
MGC10485 hypothetical protein MGC10485
MGC4170
MGC4170 protein
MGC61571 hypothetical protein MGC61571
NMUR1
neuromedin U receptor 1
OSBPL5
oxysterol binding protein-like 5
PYHIN1
pyrin and HIN domain family, member 1
SYTL3
synaptotagmin-like 3
UHMK1
U2AF homology motif (UHM) kinase 1
USP28
ubiquitin specific protease 28
UTS2
urotensin 2
Walzer et al., Curr. Opin. Immunol. 2007
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Natural Cytotoxicity Receptors
(NCR)
Expression pattern of human NKp46
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Les anti-NKp46
permettent
maintenant de
visualiser les cellules
NK sur coupe
Spleen
Walzer et al. PNAS, 2007
Gregoire et al, Immunol. Rev, 2007
NK cells in the skin and gut human
Luci, Reynders et al, Nat Immunol, 2009
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NK cells in the skin and gut mouse
Luci, Reynders et al, Nat Immunol, 2009
NK cells are present in all
sections of the gut
Luci, Reynders et al, Nat Immunol, 2009
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Phenotype and functions of
mucosal « NK cells »
-Mucosal NK cells do not express T-bet but express RORgt
-Mucosal NK cells are not cytotoxic and do not produce IFNg but produce IL22 (but not IL17) in response to stimulation by IL23
-Mucosal NK cells are developmentally related to Lti cells
ÆAre mucosal NK cells real NK cells? ÆNCR22 cells
Developmental relationship between
classical NK cells and NCR22 cells
NK
NCR22
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IL-22
• Protection et maintenance des barrières
épithéliales
– Induit la survie des cellules épithéliales
– Induit la production de défensines antibactériennes
• Æprotège contre certaines bactéries (Citrobacter
Rodentium)
• Æprotège contre des maladies inflammatoires
(hépatites, myocardite autoimmune, IBD)
• Æprotège dans les allergies de la peau?
NK, NCR22 and skin pathologies
• Very few data on NCR22
• Classical NK detected in many skin
allergies/inflammation but their role is not
clear
• NK cells and DNFB-induced contact
hypersensitivity (group of U. Von Andrian,
Boston)
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Problèmes
• CHS dans les souris RAG KO pas
reproduite dans deux labos différents
• Pas de récepteur identifié
• Les cellules NK « mémoire » se
localiseraient pour une raison inconnue
dans le foie
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