Rôle des cellules NK dans l`immunité de la peau et des muqueuses
Transcription
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 [email protected] INSERM U851 Lyon Innate Immunity Effector Functions « Pathogens » Initiation of Ag-specific response Effector Functions Adaptive Immunity 1 22/02/2011 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)+ 2 22/02/2011 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 3 22/02/2011 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 4 22/02/2011 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 5 22/02/2011 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 6 22/02/2011 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 7 22/02/2011 Evolutionary success: • High transmissibility • Low pathogenicity • Persist in a latent state 8 22/02/2011 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 ») 9 22/02/2011 Case of CMV But CMV downregulates NKG2D ligands 10 22/02/2011 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. 11 22/02/2011 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 12 22/02/2011 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 13 22/02/2011 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 14 22/02/2011 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+? 15 22/02/2011 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 16 22/02/2011 Natural Cytotoxicity Receptors (NCR) Expression pattern of human NKp46 17 22/02/2011 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 18 22/02/2011 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 19 22/02/2011 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 20 22/02/2011 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) 21 22/02/2011 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 22