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Graft-versus-host disease is enhanced by extracellular ATP activating P2X7R

Abstract

Danger signals released upon cell damage can cause excessive immune-mediated tissue destruction such as that found in acute graft-versus-host disease (GVHD), allograft rejection and systemic inflammatory response syndrome. Given that ATP is found in small concentrations in the extracellular space under physiological conditions1, and its receptor P2X7R is expressed on several immune cell types, ATP could function as a danger signal when released from dying cells. We observed increased ATP concentrations in the peritoneal fluid after total body irradiation, and during the development of GVHD in mice and in humans. Stimulation of antigen-presenting cells (APCs) with ATP led to increased expression of CD80 and CD86 in vitro and in vivo and actuated a cascade of proinflammatory events, including signal transducer and activator of transcription-1 (STAT1) phosphorylation, interferon-γ (IFN-γ) production and donor T cell expansion, whereas regulatory T cell numbers were reduced. P2X7R expression increased when GVHD evolved, rendering APCs more responsive to the detrimental effects of ATP, thereby providing positive feedback signals. ATP neutralization, early P2X7R blockade or genetic deficiency of P2X7R during GVHD development improved survival without immune paralysis. These data have major implications for transplantation medicine, as pharmacological interference with danger signals that act via P2X7R could lead to the development of tolerance without the need for intensive immunosuppression.

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Figure 1: ATP abundance is increased in ascites of individuals developing GVHD and in the mouse gastrointestinal tract.
Figure 2: P2R signaling blockade and neutralization of ATP reduces GVHD severity in vivo.
Figure 3: Impact of ATP and P2R signaling blockade on CD80 and CD86 expression, T cell proliferation and cytokine production.
Figure 4: P2X7R deficiency of recipient DCs leads to GVHD protection.

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References

  1. Zimmermann, H. Extracellular metabolism of ATP and other nucleotides. Naunyn Schmiedebergs Arch. Pharmacol. 362, 299–309 (2000).

    Article  CAS  Google Scholar 

  2. Di Virgilio, F. Purinergic mechanism in the immune system: A signal of danger for dendritic cells. Purinergic Signal. 1, 205–209 (2005).

    Article  CAS  Google Scholar 

  3. Granstein, R.D. et al. Augmentation of cutaneous immune responses by ATP γ S: purinergic agonists define a novel class of immunologic adjuvants. J. Immunol. 174, 7725–7731 (2005).

    Article  CAS  Google Scholar 

  4. Pellegatti, P. et al. Increased level of extracellular ATP at tumor sites: in vivo imaging with plasma membrane luciferase. PLoS ONE 3, e2599 (2008).

    Article  Google Scholar 

  5. Anderson, B.E. et al. Effects of donor T cell trafficking and priming site on graft-versus-host disease induction by naive and memory phenotype CD4 T cells. Blood 111, 5242–5251 (2008).

    Article  CAS  Google Scholar 

  6. Hill, G.R. & Ferrara, J.L. The primacy of the gastrointestinal tract as a target organ of acute graft-versus-host disease: rationale for the use of cytokine shields in allogeneic bone marrow transplantation. Blood 95, 2754–2759 (2000).

    CAS  PubMed  Google Scholar 

  7. Heiss, K. et al. High sensitivity of intestinal CD8+ T cells to nucleotides indicates P2X7 as a regulator for intestinal T cell responses. J. Immunol. 181, 3861–3869 (2008).

    Article  CAS  Google Scholar 

  8. Hauri-Hohl, M.M. et al. Donor T cell alloreactivity against host thymic epithelium limits T cell development after bone marrow transplantation. Blood 109, 4080–4088 (2007).

    Article  CAS  Google Scholar 

  9. Pannetier, C. et al. The sizes of the CDR3 hypervariable regions of the murine T cell receptor β chains vary as a function of the recombined germ-line segments. Proc. Natl. Acad. Sci. USA 90, 4319–4323 (1993).

    Article  CAS  Google Scholar 

  10. Zeiser, R. et al. Host-derived interleukin-18 differentially impacts regulatory and conventional T cell expansion during acute graft-versus-host disease. Biol. Blood Marrow Transplant. 13, 1427–1438 (2007).

    Article  CAS  Google Scholar 

  11. Burchill, M.A., Yang, J., Vogtenhuber, C., Blazar, B.R. & Farrar, M.A. IL-2 receptor β–dependent STAT5 activation is required for the development of Foxp3+ regulatory T cells. J. Immunol. 178, 280–290 (2007).

    Article  CAS  Google Scholar 

  12. Vogtenhuber, C. et al. Constitutively active Stat5b in CD4+ T cells inhibits graft-versus-host disease (GVHD) lethality associated with increased regulatory T cell (Treg) potency and decreased T effector cell (Teff) responses. Blood 116, 466–474 (2010).

    Article  CAS  Google Scholar 

  13. Zeiser, R. et al. HMG-CoA reductase inhibitors (statins) provide acute-graft-versus-host disease protection by TH-2 cytokine induction while sparing graft-versus-leukemia activity. Blood 110, 4588–4598 (2007).

    Article  CAS  Google Scholar 

  14. Wang, H. et al. Paradoxical effects of IFN-γ in graft-versus-host disease reflect promotion of lymphohematopoietic graft-versus-host reactions and inhibition of epithelial tissue injury. Blood 113, 3612–3619 (2009).

    Article  CAS  Google Scholar 

  15. Sadzak, I. et al. Recruitment of Stat1 to chromatin is required for interferon-induced serine phosphorylation of Stat1 transactivation domain. Proc. Natl. Acad. Sci. USA 105, 8944–8949 (2008).

    Article  CAS  Google Scholar 

  16. Solle, M. et al. Altered cytokine production in mice lacking P2X7 receptors. J. Biol. Chem. 276, 125–132 (2001).

    Article  CAS  Google Scholar 

  17. Ferrari, D. et al. The P2X7 receptor: a key player in IL-1 processing and release. J. Immunol. 176, 3877–3883 (2006).

    Article  CAS  Google Scholar 

  18. Rao, V., Saunes, M., Jørstad, S. & Moen, T. In vitro experiments demonstrate that monocytes and dendritic cells are rendered apoptotic by extracorporeal photochemotherapy, but exhibit unaffected surviving and maturing capacity after 30 Gy gamma irradiation. Scand. J. Immunol. 68, 645–651 (2008).

    Article  CAS  Google Scholar 

  19. Kawamura, H. et al. P2X7 receptor–dependent and –independent T cell death is induced by nicotinamide adenine inucleotide. J. Immunol. 174, 1971–1979 (2006).

    Article  Google Scholar 

  20. Aswad, F., Kawamura, H. & Dennert, G. High sensitivity of CD4+CD25+ T cells to extracellular metabolites nicotinamide adenine dinucleotide and adenosine triphosphate: a role for P2X7 receptors. J. Immunol. 175, 3075–3083 (2005).

    Article  CAS  Google Scholar 

  21. Chen, L. & Brosnan, C.F. Regulation of immune response by P2X7 receptor. Crit. Rev. Immunol. 26, 499–513 (2006).

    Article  CAS  Google Scholar 

  22. Kanneganti, T.D. et al. Pannexin-1–mediated recognition of bacterial molecules activates the cryopyrin inflammasome independent of Toll-like receptor signaling. Immunity 26, 433–443 (2007).

    Article  CAS  Google Scholar 

  23. Hill, G.R. et al. Differential roles of IL-1 and TNF-α on graft-versus-host disease and graft versus leukemia. J. Clin. Invest. 104, 459–467 (1999).

    Article  CAS  Google Scholar 

  24. Lee, K.H. et al. P2X7 receptor polymorphism and clinical outcomes in HLA-matched sibling allogeneic hematopoietic stem cell transplantation. Haematologica 92, 651–657 (2007).

    Article  CAS  Google Scholar 

  25. Metaxas, Y. et al. Human hematopoietic cell transplantation results in generation of donor-derived epithelial cells. Leukemia 19, 1287–1289 (2005).

    Article  CAS  Google Scholar 

  26. Cao, Y.A. et al. Shifting foci of hematopoiesis during reconstitution from single stem cells. Proc. Natl. Acad. Sci. USA 101, 221–226 (2004).

    Article  CAS  Google Scholar 

  27. Reichardt, W. et al. Impact of mammalian target of rapamycin inhibition on lymphoid homing and tolerogenic function of nanoparticle-labeled dendritic cells following allogeneic hematopoietic cell transplantation. J. Immunol. 181, 4770–4779 (2008).

    Article  CAS  Google Scholar 

  28. Zeiser, R. et al. Differential impact of mTOR inhibition on CD4+CD25+Foxp3+ regulatory T cells as compared to conventional CD4+ T cells. Blood 111, 453–462 (2008).

    Article  CAS  Google Scholar 

  29. Kaplan, D.H. et al. Target antigens determine graft-versus-host disease phenotype. J. Immunol. 173, 5467–5475 (2004).

    Article  CAS  Google Scholar 

  30. Krempl, C.D. et al. Identification of a novel virulence factor in recombinant pneumonia virus of mice. J. Virol. 81, 9490–9501 (2007).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We are grateful to S. Krüger, C. Faller, C. Kiesel, C.A. Bäuerlein, M. Ritz and V. Schmidt. This study was supported by the Forschungskommission der Universität Freiburg (FoKo ZEI616/08 to R.Z.), the Wilhelm Sander Stiftung (2008.046.1 to R.Z., M.I. and J.F.) and the Deutsche Forschungsgemeinschaft (ID 7/4-1 to M.I. and ZE 872/1-1 to R.Z.).

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K.W., J.G. and T.M. helped to design the experiments, performed experiments, analyzed data and helped to write the manuscript. C.D. and M.G. helped with experiments. A.B. and C.D.K. designed, performed and analyzed the PVM experiments and helped to write the manuscript. S.S. helped to design the project and experiments and helped to write the manuscript. U.V.G., E.J. and N.K. performed histopathology scoring and helped to write the manuscript. A.Z. helped to collect clinical samples and medical history and helped to write the manuscript. F.G. performed flow cytometry analysis for P2X7R of human peripheral blood mononuclear cells (PBMCs) and helped to write the manuscript. P.P., F.D.V. and D.F. provided the PME cells, helped to design the in vivo ATP detection studies and helped to write the manuscript. P.F. helped to design and supervise spectratyping experiments and helped to write the manuscript. J.F. helped to design the project and experiments and helped to write the manuscript. M.I. and R.Z. designed the studies, shared the supervision of the work, analyzed data and wrote the manuscript. All authors have read and agreed to the final version of the manuscript.

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Correspondence to Marco Idzko or Robert Zeiser.

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The authors declare no competing financial interests.

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Supplementary Figures 1–4, Supplementary Tables 1 and 2 and Supplementary Methods (PDF 659 kb)

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Wilhelm, K., Ganesan, J., Müller, T. et al. Graft-versus-host disease is enhanced by extracellular ATP activating P2X7R. Nat Med 16, 1434–1438 (2010). https://doi.org/10.1038/nm.2242

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