1 / 55

SIXTH INTERNATIONAL SYMPOSIUM ON STEM CELL THERAPY AND CARDIOVASCULAR INNOVATIONS

SIXTH INTERNATIONAL SYMPOSIUM ON STEM CELL THERAPY AND CARDIOVASCULAR INNOVATIONS. Madrid , April 23th-24th , 2009. IS IT TIME TO USE ALLOGENEIC STEM CELLS?. AN IMMUNOLOGICAL PERSPECTIVE. Dominique CHARRON, MD-PhD (Dominique.Charron@sls.aphp.fr)

zeno
Download Presentation

SIXTH INTERNATIONAL SYMPOSIUM ON STEM CELL THERAPY AND CARDIOVASCULAR INNOVATIONS

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. SIXTH INTERNATIONAL SYMPOSIUM ON STEM CELL THERAPY AND CARDIOVASCULAR INNOVATIONS Madrid, April23th-24th, 2009 IS IT TIME TO USE ALLOGENEIC STEM CELLS? AN IMMUNOLOGICAL PERSPECTIVE Dominique CHARRON, MD-PhD (Dominique.Charron@sls.aphp.fr) Hôpital Saint Louis, IUH - INSERM U 940 – CIB-HOG – Université Paris Diderot

  2. ALLOGENIC IMMUNITY I: MOLECULAR BASIS HLA MICA

  3. Nat.Rev.Cancer.2004. 4:371-380

  4. ALLOGENEICIMMUNITYII : CELLULAR RESPONSES INDUCTION/TRIGGERING/ EFFECTOR PHASE (REJECTION) Preexisting Allogeneic T Cells (CD8/CD4) Immediate Response 1- 5 % of Circulating T Cells vs< 0.5% for Ag 2 pathways Direct – Indirect

  5. Tc Cytotoxic T-cell Th Helper T-cell Allo MHC molecules from the Donor are recognized by Host T-cells Class I Class II Allogeneic (Donor) APC (stimulator) Allogeneic (Donor) Cell MHC or other molecules are shed taken up and processed by host APC Tc Cytotoxic T-cell Direct allorecognition Th Helper T-cell Host APC (recipient) Pathways of allorecognition Peptide derived from allo molecules presented on host MHC to Host T-cells Indirect allorecognition

  6. IMMUNE RECOGNITION: SELF VS NON SELF VS ALLO + SELECTION - MHC RESTRICTION - SELECTION - SELFTOLERANCE • ALLO RECOGNITION – ALLO RESPONSES • 5 % • ALLO CMH = SELF + X (Ag)

  7. ALLOGENEIC SC ARE NOT IMMUNO PRIVILEGED !!! THREE SUPPORTING PAPERS • MHC EXPRESSION • IMMUNOGENEICITY INCREASES UPON DIFFERENCIATION • IN VIVO REJECTION

  8. 1 Characterization of the expression of MHC proteins in human embryonic stem cells Micha Drukker, Gil Katz, Achia Urbach, Maya Schuldiner, Gal Markel, Joseph Itskovitz-Eldor, Benjamin Reubinoff, Ofer Mandelboim, and Nissim Benvenisty PNAS, 2002, 99:9864

  9. b2m HLA-I Counts 721.221 HLA-II Fluorescence intensity Expression of MHC proteins in undifferentiated and differentiated human ES cells

  10. IFN-g induction of MHC-I in human ES cells is dose and time dependent

  11. 2 Embryonic Stem Cell Immunogenicity Increases Upon Differentiation After Transplantation Into Ischemic Myocardium Rutger-Jan Swijnenburg, MS; Masashi Tanaka, MD; Hannes Vogel, MD; Jeanette Baker, PhD; Theo Kofidis, MD; Feny Gunawan, BS; Darren R. Lebl, MS; Anthony D. Caffarelli, MD; Jorg L. de Bruin, MD; Eugenia V. Fedoseyeva, PhD; Robert C. Robbins, MD Circulation. 2005;112:I-166-I-172

  12. Graft infiltration of immune cells after transplantation of in vivo differentiated ESCs

  13. 3 Immunosuppressive therapy mitigates immunological rejection of human embryonic stem cell xenografts Rutger-Jan Swijnenburg, Sonja Schrepfer, Johannes A. Govaert, Feng Cao, Katie Ransohoff, Ahmad Y. Sheikh, Munif Haddad, Andrew J. Connolly, Mark M. Davis, Robert C. Robbins, and Joseph C. Wu PNAS, 2008,105:12991

  14. In vivo visualization of hESC survival

  15. MEDICAL CONSEQUENCES OF ALLORECOGNITION

  16. Multivariate Analysis (Cox) 1,78* Failure : RR 1,5 1,61* 1,37 1,37 1 1,12 RR 0 143 1 655 2 2015 3 3288 4 2576 5 1049 6 177 1 1 Incompatibilities HLA-A-B-DR HLA incompatibility (Ab = 0%)

  17. Three-year graft survival rates (A) and frequency of rejection treatment during first year (B) in relation to number of HLA-A, -B, and -DR mismatches in patients transplanted from 2000 to 2004. p<0.001

  18. PRE IMPLANT Ab (TRANSFUSION/PREGNANCY) Ab DETECTION CROSS MATCH

  19. Terasaki et al. Predictive Value of HLA Antibodies and Serum Creatinine in Chronic Rejection: Results of a 2-year Prospective Trial Transplantation 2005;80: 1194–1197

  20. De Novo ANTI HLA ANTIBODIES (+ CD 4d deposit) DEVELOP AFTER RENAL TRANSPLANTATION PROLONGED ABSENCE OF ANTIBODY CORRELATES WITH GOOD FUNCTION WHILE 86% OF GRAFT FAILURE HAVE ANTI HLA AND/OR MICA Abs PROSPECTIVE STUDY HEART LUNG

  21. 1910 RENAL TRANSPLANTATION ANTIBODIES AGAINST MICA ANTIGENS 217 + (11.4%) 1 YEAR ALLOGRAFT SURVIVAL ALL CASES 88.3 vs 93 p0.01 FIRST TRANSPLANT 87 vs 93.5 p0.005 WELL HLA MATCHED 83.2 vs 95.1 p0.002 Zav et al. NEJM 357, 1293, 2008

  22. HSCT HLA MATCHED SIBLINGS GVHD 30 % UP PERFECT HLA MATCH # OPTIMAL GENETIC MAKE UP OUTCOMES GENES GVHD - REJECTION INFECTIONS TRM – RELAPSE SURVIVAL HLA NON HLA MHC NON MHC INTEGRATED GENETICS ADAPTED TREATMENT DONOR SELECTION PREVENTION / PROPHYLAXIS

  23. Unrelated BMT Survivalaccording to number of HLA disparities 11 (n=48) 11-12 (n=62) 12 (n=14) 10 (n=34) 6-10 (n=59) 6-9 (n=25)

  24. Risk of of severe bacterial infections: HLA-E*0101/0101 donor genotype: 5318% vs 2512; PGray test= 0.035 Risk of transplant-related mortality: HLA-E*0101/0101 donor genotype: 5318% vs 2714% PGray test= 0.08

  25. CYTOKINES - INFLAMMATION

  26. HSCT : INNATE IMMUNOGENETICS Frequency/Severity of infectious (viral, bacterial, fungal) events Intricated with GVHD TRM - Survival INFECTIONS/GVHD TRM GENES GENOTYPES • MPO AG or AA (non CC) • MBL MBL2 • Fc RII a (CD 32) R 131 • Fc RIII b HNA-1a • TLR 4 - • NOD2/CARD 15 SNP 8, 12, 13 • Oestrogen Receptor intron 1 • Vitamin D Receptor intron 8 • Fas - 670 G

  27. STEM CELL/RECIPIENT COMPATIBILITY PREDICTION ANTI- HLA/MIC A DETECTION (quarterly),IDENTIFICATION + CROSS MATCHING PRE TRANSPLANT (FINAL TEST) Best acheviable HLA A, B, DR, DQ, Typing C, DP mandatory if antibodies present Post transplantation FOLLOW UP Donor specific Ab HLA - MIC

  28. IMMUNOGENETIC MATCHING STEM CELL BANKS ABO COMPATIBILITY feasible + HLA COMPATIBILITY Level 1 Generic matching (OT) Level 2 Intermediate matching (CB) Level 3 Full matching (UD BMT) + m Hag COMPATIBILITY !!!

  29. LOOKING FORWARD  MATCHING - REDUCING MISMATCHING • STEM CELL BANKS • STEM CELL INGENEERING • IMMUNOGENETIC SELECTION • INNOVATIVE IMMUNOSUPPRESSION/INDUCTION OF TOLERANCE T CELLS COSTIMULATION BLOCKADE REGULATORY T CELLS B CELLS IV Ig CD 40 L BY ANTI CD 40 L B7 BY CTLA 4 Ig ANTI LFA1… CD 20 VELCADE • GENERATION OF MIXED CHIMERISM/ THYMIC INGENEERING • STANDARD CHRONIC IMMUNO SUPPRESSION: • INFECTIONS/CANCER, AGENT SPECIFIC KIDNEY FAILURE, DIABETES, OSTEOPOROSIS, HYPERTENSION…

  30. THE STORY BOARD OF MEDICINE PREDICTIVE MEDICINE MYTH ANTICIPATIVE GENETICS/ PHARMACOGENETICS INDIVIDUALIZED SUSCEPTIBILITY GOD OF HEALING 1st HOSPITAL PREVENTIVE MEDICINE MEDICINE HIPPOCRATES OF KOS HOLISTIC PUBLIC HEALTH INDIVIDUAL + POPULATION INTEGRATIVE 1st SCHOOL OF MEDICINE EMPIRISM ECONOMICS SCIENCE

  31. Immunogenicity and Allogenicity: A Challenge of Stem Cell Therapy Dominique Charron.Caroline Suberbielle-Boissel.Reem Al-Daccak J.of Cardiovasc.Trans.Res.2009,8- 9062-9

  32. AGING OF HSC

  33. Aging and HSC • Increased number of HSC with age in certain strains of mice (Morrison et al 1996, Nature Med) • Increased number of HSC in cycle (Rossi et al 2005, PNAS) • Expression of senescence marker (p16) (Janzen et al 2006, Nature,) • Decreased reconstitution capacity - lymphoid compartment. (Sudo et al 2000, JEM) • Intrinsic changes in the lymphoid potential of HSC ?

  34. Quantitative in vitro analysis of HSC differentiation potential Old 18-24m Young 1-2m Bone marrow CLPfraction HSC fraction (Lin-Sca1hicKithiIL7Ra-) (Lin-Sca1locKitloIL7Ra+) Plate in 96 wells at limiting dilution OP9 + IL7 OP9 + IL7 Facs sort wells • Frequency CD19+ (B cells) • Frequency CD11b+ (Myeloid)

  35. HSC : decreased B but not myeloidpotentialwithage Old Old Young Young

  36. IRES IRES CONT +EBF CONT +Pax5 LTR LTR LTR LTR P P GFP GFP Pax5 EBF IRES CONT LTR LTR P GFP Transduction of EBF and Pax5 in CLP fromoldmice Young (1-2m) Old (18-24m) Isolate CLP Lin-IL-7R+Sca-1+c-Kit+ Transduction CONT±EBF,Pax5 Measure B cell potential In vitrodifferentiation OP9 + IL-7, c-KitL, Flt3 Frequency of B cell clones (CD19+)

  37. IRES +EBF Young (1-2m) Old (18-24m) LTR LTR P GFP EBF Isolate CLP Lin-IL-7R+Sca-1+c- Kit+ Transduction with EBF vector Measure B cell potential In vitrodifferentiation OP9 + IL-7, c-KitL, Flt3 Frequency of B cell clones (CD19+) EBF expression restores B cell potential of CLP from old mice IRES CONT LTR LTR P GFP B potential Old 1/570 % negative wells Old+EBF 1/43 1/18 Young Number of cells / well

  38. Young (1-2m) Old (18-24m) Isolate CLP Lin-IL-7R+Sca-1+c-Kit+ Transduction with PAX5 vector Measure B cell potential In vitrodifferentiation OP9 + IL-7, c-KitL, Flt3 Frequency of B cell clones (CD19+) Pax5 expression restores B cellpotential of CLP fromoldmice IRES +Pax5 LTR LTR P GFP Pax5 IRES CONT LTR LTR P GFP B Potential Old 1/285 % negative wells Old+Pax5 1/41 1/9 Young Number of cells / well

  39. Conclusion T/NK prgenitor Aging immature B cell HSC CLP Pro-B Pre-B EBF Pax5 • Intrinsicloss of B cell potential of HSC with age • Due to decreased expression of EBF and Pax5

  40. IMMUNOSCENESCENCE • DECREASES LYMPHOPOEISIS • THYMIC INVOLUTION AND DECLINE IN T CELL FUNCTION • SUBSTANTIAL CHANGES IN AN B CELL COMPARTMENTS • AGE-RELATED HSC DEFECT ??

  41. THE IMMUNE CLOCK

  42. Nat.Rev.Cancer.2004. 4:371-380

  43. Fifth International Symposium on Stem CellTherapy and AppliedCardiovascularBiotechnology MADRID, April 24th-25th, 2008 « IMMUNITY AND ALLOGENEICITY : STEMCELLNESS EDUCATION VS AND DIVERSITY » Dominique CHARRON, MD-PhD(Dominique.Charron@sls.aphp.fr) Hôpital Saint Louis, IUH - INSERM U 662 – CIB-HOG – Université Paris Diderot UNIVERSITE DE PARIS 7 DENIS DIDEROT

More Related