DQ8-villin-IL-15tg

SYNERGISTIC ROLE OF IL-15IN THE EPITHELIUM AND THE LAMINA PROPRIAIN CELIAC DISEASE PATHOGENESISBana JabriDepartment of MedicineCeliac Disease CenterUniversity of Chicago

IngestedglutenEnvironmentalfactorsgliadinVRVPVPQLQPQNPSQQQPQEQVPLVQQQQFLGQQQPFPPQQPYPQPQPFPSQQPYLQLQPFPQPQLPYSQPQPFRPQQPYGluten is not digested(proline-rich)protease-resistantlong gliadinpeptidesShan et al., Science, 2002(glutamine-rich)Tissue Transglutaminase (TG2)LQLQPFPQPQLPYPQPQLPYPQPQLPYPQPQPFVader et al., J exp Med, 2002Molberg et al., Nat Med, 1998Van de Wal et al., J immunol, 1998TG2QQYPSGQGSFQPSQQNPQPQPELPYPQPPFPQPELPYPimmunogenic gliadin peptidesEGSFQPSQETCRHLA-DQ2CD4CD4TCRHLA-DQ8Vader et al., J exp Med, 2002Molberg et al., Nat Med, 1998Van de Wal et al., J immunol, 1998anti-gluten T cell responseNilsen et al., Gut, 1995Molberg et al., Scand.J.Immunol., 1997Nilsen et al., Gastroenterology, 1998

Intestinal HomeostasisDefault response in the gut to oral antigen: REGULATORY IMMUNE RESPONSEGut lumenFood antigenGut floraGlutenEpitheliumEffectorT cellsLaminapropriaORALTOLERANCEIFN-γIL-21CELIACDISEASETGF-βTreg Treg inducible Treg cellsRA?T H 1Mesentericlymph nodeNaïveCD4 T cell

IL-15 is over-expressed in the lamina propria of active celiacdisease patientsMaiuri L. et al., Gastroenterology 2000Mention J.J. et al. Gastroenterology 2003DePaolo R.W., Abadie V. et al., Nature 2011

Is there a link between dysregulated IL-15 expression andthe loss of tolerance to oral antigens?

D d -IL-15 transgenic mouse model reproducesthe high levels of IL-15 expression observed in the lamina propriaof active celiac disease patientsD d -IL-15tg D d promoter IL-2 leader peptide IL-15 FLAG hGHFehniger T.A. et al., J.Exp.Med. 2001

IL-15 induces an inflammatory IFN-γ + anti-gliadin T cell responseafter oral ingestion of gliadin

IL-15 promotes a weak T H 17 anti-gluten T cell responseafter oral ingestion of gliadinCeliac DiseaseCrohn’s diseaseHunt, K. A. et al. Nat. Genet., 2008Franke A. et al., Nat. Genet., 2010

Perspectives: Blocking IL-15 signaling prevents theinduction of inflammatory T cell responses to fed antigenDQ8/ DQ8-D d -IL-15tg miceanti-IL-15TMβ-1 (anti-IL-2Rβ)anti-IL-12i.p.PBS (sham)gliadini.g.10 daysRe-stimulation of Lp and MLN cellsW/gliadinAnalysis of IFN-γ production

Intraepithelial lymphocytosis, anti-gliadin and anti-transglutaminaseIgG and IgA in in gliadin fed DQ8-D d -IL-15tg miceDQ8DQ8-D d -IL-15tg

DQ8-D d -IL-15tg mice fed gluten phenocopythe early stages of Celiac Disease characterized by the inductionof an adaptive anti-gluten immunity in the absence of villous atrophy(Potential Celiac Disease)• IFN-γ + and IL-21 + anti-gluten CD4 + T cell responses• Anti-gliadin antibodies•Autoantibodies: Anti-transglutaminases•Infiltration of intraepithelial lymphocytesMarsh 1Marsh 1DePaolo R.W. & Abadie V.et al., Nature, 2011

The celiac icebergVillous atrophy+/- clinical manifestations?Potential CDAnti-TTG and normal histology

Effector intraepithelial CTL killstressed epithelial cellsCTLCTLgliadinαβ TCRαβ TCRInduction of activatingnatural killer receptorsInduction of ligands foractivating natural killerIECgliadinIECstressDirect recognitionof gliadin peptidesIndirect gluten“stress-mediated” response

Healthy tissueIntraepithelialcytotoxic T cell (IE-CTL)Epithelial cellStressed or inflamed tissueCD94/NKG2AcytolysisInhibitionTCRCD94/NKG2C HLA-EIFN-γStressLaminapropriaNKG2DNKG2DAAMICA/BIL-15AALeukotrienes?APCNeutrophilTissue maintenanceGlutenMicrobiotaSelf-antigenGlutenMicrobiotaSelf-antigenEosinophilTissue damageMesentericlymph nodeNKG2DMeresse et al., Immunity 2004TCRNaïve CD8 + T cellMemory CTLHue et al., Immunity 2004Meresse et al., J Exp Med 2006Tang et al., J Exp Med 2009

INNATE IMMUNE ACTIVATION“CD4 anti-gluten T cell responses are requiredbut not sufficient to induce intestinal damage”Adaptive immune response(anti-TG2 antibodies)Innate epithelial response(IL-15, epithelial stress markers)Kaukinen et al. Scand J Gastroenterol, 2005Marietta et al. JCI, 2004de Kauwe et al., J Immunol, 2009IEL activation↓Inhibitory NKG2A↑Activatory NKR (CD94+NKG2A-, NKG2D+)Jabri et al., Gastroenterology 2001Meresse et al., Immunity 2004Hue et al., Immunity 2004Meresse et al., J.Exp.Med. 2006VILLOUS ATROPHYInhibitory NK receptorsActivatory NK receptorsActivatory NK receptorsSetty M. et al., in preparation

DQ8-D d -IL-15tgDQ8-villin-IL-15tgDQ8-villin-D d -IL-15tg=“Triple tg”D d promoterIL-15D d promoterIL-15villin promoterIL-15Xvillin promoterIL-15PTD-gliadin30 daysAnalysis of NK receptors on IELsHistologyDQ8DQ8-D d -IL-15tgDQ8-villin-IL-15tgDQ8-D d -villin-IL-15tg

SHAMDQ8 DQ8-D d -IL-15tg DQ8-villin-IL-15tg DQ8-D d -villin-IL-15tg

DQ8-D d -villin-IL-15tg mice fed gliadin have increased amounts ofactivatory NK receptors expressing CD8 T cells400000p=0.0002p=0.0003p=0.0512p=0.05743000002000001000000

DQ8-D d -villin-IL-15tg mice fed gliadin have increased amounts ofCD8 T cells expressing activating NK receptors

6NSp=0.0031DQ8-D d -villin-IL-15tg mice fed glutendevelop villous atrophy543NS2DQ8-D d -IL-15tgDQ8-villin-IL-15tgDQ8-villin-D d -IL-15tgControl(sham)GliadinBrdU

In absence of DQ8D d -villin-IL-15tg mice fed gliadindo not expand IE-CTL with activating NK receptorsdo not develop villous atrophyDQ8-D d -villin-IL-15tgD d -villin-IL-15tgShamGliadin

IL-15 IN THE LAMINA PROPRIA? ?Role of DQ8Role of TG2?IL-15 IN THE EPITHELIUMAdaptive immune response(anti-TG2 antibodies)Innate epithelial response(IL-15, epithelial stress markers)IEL activation↓Inhibitory NKG2A↑Activatory NKR (CD94+NKG2A-, NKG2D+)VILLOUS ATROPHY

AcknowledgmentsValerie AbadieWilliam R. DePaoloFangming TangCezary CiszewskiHannah Fehlner-PeachWenxia WangMala SettySarah KamasawkiBen SallyValentina DiscepoloStefano GuandaliniSonia KupferCarol SemradJ. TurnerMayo clinic, RochesterEric MariettaJoe MurrayUniversity Frederico II, NaplesR. TronconeS . AuricchioV.M. SalvatiValentina DiscepoloColumbia Presbyterian HospitalP.HR GreenG. BhagatNIH, BethesdaY. BelkaidJ. HallT.A. Waldmann

DQ8 DQ8-D d -IL-15tg DQ8-villin-IL-15tg DQ8-D d -villin-IL-15tgGLIADINSHAM