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Prevention of graft-versus-host-disease with preserved graft-versus-leukemia-effect by ex vivo and in vivo modulation of CD4+ T-cells

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Abstract

This is the first report showing that an epitope-specific ex vivo modulation of an allogeneic hematopoietic stem cell graft by the anti-human CD4 antibody MAX.16H5 IgG1 simultaneously facilitates the anti-tumor capacity of the graft (Graft-versus-leukemia effect, GvL) and the long-term suppression of the deleterious side effect Graft-versus-host-disease (GvHD). To distinguish and consolidate GvL from GvHD, the anti-human CD4 antibody MAX16.H5 IgG1 was tested in murine GvHD and tumor models. The survival rate was significantly increased in recipients receiving a MAX.16H5 IgG1 short-term (2 h) pre-incubated graft even when tumor cells were co-transplanted or when recipient mice were treated by MAX.16H5 IgG1 before transplantation. After engraftment, regulatory T-cells are generated only supporting the GvL effect. It was also possible to transfer the immune tolerance from GvHD-free recipient chimeras into third party recipient mice without the need of reapplication of MAX.16H5 IgG1 anti-human CD4 antibodies. These findings are also benefical for patients with leukemia when no matched related or unrelated donor is available and provides a safer allogeneic HSCT, which is more effective against leukemia. It also facilitates allogeneic (stem) cell transplantations for other indications (e.g., autoimmune-disorders).

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Abbreviations

APCs:

Antigen-presenting cells

BM:

Bone marrow

FoxP3:

Forkhead box P3

GvHD:

Graft-versus-host-disease

GvL:

Graft-versus-leukemia

HLA:

Human leukocyte antigen

HSCT:

Hematopoietic stem cell transplantation

IgG:

Immunoglobulin G

MHC:

Major histocompatibility complex

PCR:

Polymerase chain reaction

SpC:

Splenocytes

TTG mice:

Triple transgenic mice

WBC:

White blood cell count

References

  1. Jacobsohn DA, Vogelsang GB (2007) Acute graft versus host disease. Orphan J Rare Dis 2:35

    Article  Google Scholar 

  2. Ferrara JL, Levine JE, Reddy P, Holler E (2009) Graft-versus-host disease. Lancet 373(9674):1550–1561

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  3. Hill GR, Krenger W, Ferrara JL (1997) The role of cytokines in acute graft-versus-host disease. Cyto Cell Mol Ther 3(4):257–266

    CAS  Google Scholar 

  4. Paczesny S, Choi SW, Ferrara JL (2009) Acute graft-versus-host disease: new treatment strategies. Curr Opin Hematol 16(6):427–436

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  5. von Bonin M, Stolzel F, Goedecke A, Richter K, Wuschek N, Holig K, Illmer T, Platzbecker U, Schaich M, Schetelig J, Kiani A, Ordemann R, Ehninger G, Schmitz M, Bornhauser M (2009) Treatment of refractory acute GVHD with third-party MSC expanded in platelet lysate-containing medium. B Mar Transpl 43(3):245–251

    Article  Google Scholar 

  6. Trapani JA, Smyth MJ (2002) Functional significance of the perforin/granzyme cell death pathway. Nat Rev Immunol 2(10):735–747

    Article  CAS  PubMed  Google Scholar 

  7. Michalek J, Collins RH, Durrani HP, Vaclavkova P, Ruff LE, Douek DC, Vitetta ES (2003) Definitive separation of graft-versus-leukemia- and graft-versus-host-specific CD4 + T cells by virtue of their receptor beta loci sequences. Proc Natl Acad Sci USA 100(3):1180–1184

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  8. Fehervari Z, Cooke A, Brett S, Turner J (2002) Perturbation of naive TCR transgenic T cell functional responses and upstream activation events by anti-CD4 monoclonal antibodies. Eur J Immunol 32(2):333–340

    Article  CAS  PubMed  Google Scholar 

  9. Harding S, Lipp P, Alexander DR (2002) A therapeutic CD4 monoclonal antibody inhibits TCR-zeta chain phosphorylation, zeta-associated protein of 70-kDa Tyr319 phosphorylation, and TCR internalization in primary human T cells. J Immunol 169(1):230–238

    Article  CAS  PubMed  Google Scholar 

  10. Madrenas J, Schwartz RH, Germain RN (1996) Interleukin 2 production, not the pattern of early T-cell antigen receptor-dependent tyrosine phosphorylation, controls anergy induction by both agonists and partial agonists. Proc Natl Acad Sci USA 93(18):9736–9741

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  11. Madrenas J, Germain RN (1996) Variant TCR ligands: new insights into the molecular basis of antigen-dependent signal transduction and T-cell activation. Semin Immunol 8(2):83–101

    Article  CAS  PubMed  Google Scholar 

  12. Chatenoud L, Waldmann H, Emmrich F (1995) Tolerance induction in the adult: ‘danger’ at Le Bischenberg. Immunol Today 16(3):121–123

    Article  CAS  PubMed  Google Scholar 

  13. Emmrich F, Horneff G, Becker W, Luke W, Potocnik A, Kanzy U, Kalden JR, Burmester G (1991) An anti-CD4 antibody for treatment of chronic inflammatory arthritis. Agen Act Suppl 32:165–170

    CAS  Google Scholar 

  14. Emmrich J, Seyfarth M, Fleig WE, Emmrich F (1991) Treatment of inflammatory bowel disease with anti-CD4 monoclonal antibody. Lancet 338(8766):570–571

    Article  CAS  PubMed  Google Scholar 

  15. Reinke P, Volk HD, Miller H, Neuhaus K, Fietze E, Herberger J, Herberger D, von Baehr R, Emmrich F (1991) Anti-CD4 therapy of acute rejection in long-term renal allograft recipients. Lancet 338(8768):702–703

    Article  CAS  PubMed  Google Scholar 

  16. Reinke P, Fietze E, Docke WD, Kern F, Ewert R, Volk HD (1994) Late acute rejection in long-term renal allograft recipients. Diagnostic and predictive value of circulating activated T cells. Transplantation 58(1):35–41

    Article  CAS  PubMed  Google Scholar 

  17. Reinke P, Kern F, Fietze E, Docke WD, Ewert R, Emmrich F, Volk HD (1995) Anti-CD4 monoclonal antibody therapy of late acute rejection in renal allograft recipients––CD4+ T cells play an essential role in the rejection process. Transpl Proc 27(1):859–862

    CAS  Google Scholar 

  18. Laub R, Dorsch M, Meyer D, Ermann J, Hedrich HJ, Emmrich F (2000) A multiple transgenic mouse model with a partially humanized activation pathway for helper T cell responses. J Immunol Meth 246(1–2):37–50

    Article  CAS  Google Scholar 

  19. Laub R, Dorsch M, Wenk K, Emmrich F (2001) Induction of immunologic tolerance to tetanus toxoid by anti-human CD4 in HLA-DR3(+)/human CD4(+)/murine CD4(−) multiple transgenic mice. Transpl Proc 33(3):2182–2183

    Article  CAS  Google Scholar 

  20. Laub R, Brecht R, Dorsch M, Valey U, Wenk K, Emmrich F (2002) Anti-human CD4 induces peripheral tolerance in a human CD4+, murine CD4−, HLA-DR+ advanced transgenic mouse model. J Immunol 169(6):2947–2955

    Article  CAS  PubMed  Google Scholar 

  21. Strauss G, Vignali DA, Schonrich G, Hammerling GJ (1994) Negative and positive selection by HLA-DR3(DRw17) molecules in transgenic mice. Immunogenetics 40(2):104–108

    CAS  PubMed  Google Scholar 

  22. Fricke S, Ackermann M, Stolzing A, Schimmelpfennig C, Hilger N, Jahns J, Hildebrandt G, Emmrich F, Ruschpler P, Posel C, Kamprad M, Sack U (2009) Allogeneic non-adherent bone marrow cells facilitate hematopoietic recovery but do not lead to allogeneic engraftment. PLoS ONE 4(7):e6157

    Article  PubMed Central  PubMed  Google Scholar 

  23. Fricke S, Fricke C, Oelkrug C, Hilger N, Schonfelder U, Kamprad M, Lehmann J, Boltze J, Emmrich F, Sack U (2010) Characterization of murine non-adherent bone marrow cells leading to recovery of endogenous hematopoiesis. Cell Mol Life Sci 67(23):4095–4106

    Article  CAS  PubMed  Google Scholar 

  24. Fricke S (2011) Measurement and illustration of immune interaction after stem cell transplantation. Meth Mol Biol 690:315–332

    Article  CAS  Google Scholar 

  25. Fricke S, Rothe K, Hilger N, Ackermann M, Oelkrug C, Fricke C, Schonfelder U, Niederwieser D, Emmrich F, Sack U (2012) Allogeneic bone marrow grafts with high levels of CD4(+) CD25(+) FoxP3(+) T cells can lead to engraftment failure. Cytom A 81(6):476–488

    Article  Google Scholar 

  26. Demehri S, Corbin A, Loriaux M, Druker BJ, Deininger MW (2006) Establishment of a murine model of aggressive systemic mastocytosis/mast cell leukemia. Exp Hematol 34(3):284–288

    Article  CAS  PubMed  Google Scholar 

  27. Cooke KR, Hill GR, Crawford JM, Bungard D, Brinson YS, Delmonte J Jr, Ferrara JL (1998) Tumor necrosis factor- alpha production to lipopolysaccharide stimulation by donor cells predicts the severity of experimental acute graft-versus-host disease. J Clin Inv 102(10):1882–1891

    Article  CAS  Google Scholar 

  28. Jerome KR, Vallan C, Jaggi R (2000) The tunnel assay in the diagnosis of graft-versus-host disease: caveats for interpretation. Pathology 32(3):186–190

    CAS  PubMed  Google Scholar 

  29. Hori S, Nomura T, Sakaguchi S (2003) Control of regulatory T cell development by the transcription factor Foxp3. Science 299(5609):1057–1061

    Google Scholar 

  30. Lange F, Hartl S, Ungethuem U, Kuban RJ, Hammerschmidt S, Faber S, Morawietz L, Wirtz H, Emmrich F, Krenn V, Sack U (2006) Anti-TNF effects on destructive fibroblasts depend on mechanical stress. Scand J Immunol 64(5):544–553

    Google Scholar 

  31. Pfaffl MW, Horgan GW, Dempfle L (2002) Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucl Acids Res 30(9):e36

    Article  PubMed Central  PubMed  Google Scholar 

  32. Horneff G, Sack U, Kalden JR, Emmrich F, Burmester GR (1993) Reduction of monocyte-macrophage activation markers upon anti-CD4 treatment. Decreased levels of IL-1, IL-6, neopterin and soluble CD14 in patients with rheumatoid arthritis. Clin Exp Immunol 91(2):207–213

    CAS  PubMed Central  PubMed  Google Scholar 

  33. Hiepe F, Volk HD, Apostoloff E, von Baehr R, Emmrich F (1991) Treatment of severe systemic lupus erythematosus with anti-CD4 monoclonal antibody. Lancet 338(8781):1529–1530

    Article  CAS  PubMed  Google Scholar 

  34. Niederwieser D, Lange T, Cross M, Basara N, Al-Ali H (2006) Reduced intensity conditioning (RIC) haematopoietic cell transplants in elderly patients with AML. Best Prac Res Clin Haematol 19(4):825–838

    Article  Google Scholar 

  35. Becker C, Bopp T, Jonuleit H (2012) Boosting regulatory T cell function by CD4 stimulation enters the clinic. Front Immunol 3:164

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  36. Cobbold SP (2005) T cell tolerance induced by therapeutic antibodies. Philos Trans R Soc Lond B 360(1461):1695–1705

    Article  CAS  Google Scholar 

  37. Broker BM, Tsygankov AY, Fickenscher H, Chitaev NA, Muller-Fleckenstein I, Fleckenstein B, Bolen JB, Emmrich F, Schulze-Koops H (1994) Engagement of the CD4 receptor inhibits the interleukin-2-dependent proliferation of human T cells transformed by Herpesvirus saimiri. Eur J Immunol 24(4):843–850

    Article  CAS  PubMed  Google Scholar 

  38. Horneff G, Guse AH, Schulze-Koops H, Kalden JR, Burmester GR, Emmrich F (1993) Human CD4 modulation in vivo induced by antibody treatment. Clin Immunol Immunopathol 66(1):80–90

    Article  CAS  PubMed  Google Scholar 

  39. Sawitzki B, Kieselbach B, Fisser M, Meisel C, Vogt K, Gaestel M, Lehmann M, Risch K, Grutz G, Volk HD (2004) IFN-gamma regulation in anti-CD4 antibody-induced T cell unresponsiveness. J Am Soc Nephrol 15(3):695–703

    Article  CAS  PubMed  Google Scholar 

  40. Nagahama K, Fehervari Z, Oida T, Yamaguchi T, Ogawa O, Sakaguchi S (2009) Differential control of allo-antigen-specific regulatory T cells and effector T cells by anti-CD4 and other agents in establishing transplantation tolerance. Int Immunol 21(4):379–391

    Article  CAS  PubMed  Google Scholar 

  41. Yoshida H, Hashizume M, Suzuki M, Mihara M (2011) Induction of high-dose tolerance to the rat anti-mouse IL-6 receptor antibody in NZB/NZW F1 mice. Rheumatol Int 31(11):1445–1449

    Article  CAS  PubMed  Google Scholar 

  42. de La Selle V, Riche N, Dorothe G, Bruley-Rosset M (1999) CD8+ cytotoxic T cell repertoire implicated in grafts-versus-leukemia effect in a murine bone marrow transplantation model. B Mar Transpl 23(9):951–958

    Article  Google Scholar 

  43. Teshima T, Hill GR, Pan L, Brinson YS, van den Brink MR, Cooke KR, Ferrara JL (1999) IL-11 separates graft-versus-leukemia effects from graft-versus-host disease after bone marrow transplantation. J Clin Inv 104(3):317–325

    Article  CAS  Google Scholar 

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Acknowledgments

We thank Mrs. A. Braun for proof reading the manuscript. The authors have no conflicting financial interests. We thank the colleagues from the Translational Centre for Regenerative Medicine, Universität Leipzig, for providing and breeding the triple transgenic (TTG) mice, Mrs. Ramona Blaschke. Mrs. Martina Fügenschuh and Mrs. Nadja Rudolph for preparing the histological experiments, Mrs. Jutta Jahns for preparing the irradiation of recipient mice, Mrs. Ellen Svanidze and Mrs. Manuela Ackermann for preparing the flow cytometric analysis. The work presented in this paper was funded by the German Federal Ministry of Education and Research (BMBF 0313452, PtJ-Bio, 0313909).

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Authors and Affiliations

  1. Fraunhofer Institute for Cell Therapy and Immunology (IZI), 04109, Leipzig, Germany

    Stephan Fricke, Nadja Hilger, Peter Ruschpler, Christopher Oelkrug, Ulrich Sack & Frank Emmrich

  2. Institute of Clinical Immunology, Universität Leipzig, 04109, Leipzig, Germany

    Stephan Fricke, Andreas Boldt, Ulrich Sack & Frank Emmrich

  3. Ambulante Dienste, LUPS, 6005, Luzern, Switzerland

    Christian Fricke

  4. Department of Hematology and Oncology, Leipzig University Hospital, 04109, Leipzig, Germany

    Stephan Fricke & Gerhard Behre

  5. Division of Oncology and Hematology, Evangelic Diaconic Hospital, 04177, Leipzig, Germany

    Uta Schönfelder

  6. Translational Centre for Regenerative Medicine, Universität Leipzig, 04109, Leipzig, Germany

    Stephan Fricke, Ulrich Sack & Frank Emmrich

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  1. Stephan Fricke
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Correspondence to Stephan Fricke.

Electronic supplementary material

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Supplementary Fig. 1. a–d Tolerance transfer by transplantation of 8 × 107 BM and 8 × 107 BM cells + 1 × 108 splenocytes from GvHD-free chimeric TTG-Balb/c mice into third party Balb/cwt recipient mice. BM cells (8 × 107) and BM cells + splenocytes (8 × 107 BM + 1 × 108 splenocytes) were transplanted into third party Balb/cwt mice without pre-incubation with MAX.16H5 IgG1 (n = 1, one experiment). Engrafted recipient mice of both groups showed a survival of 100 % (a), did not develop aGvHD (b), put on weight (c), and showed a stable leukocyte recovery (d).

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Fricke, S., Hilger, N., Fricke, C. et al. Prevention of graft-versus-host-disease with preserved graft-versus-leukemia-effect by ex vivo and in vivo modulation of CD4+ T-cells. Cell. Mol. Life Sci. 71, 2135–2148 (2014). https://doi.org/10.1007/s00018-013-1476-0

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