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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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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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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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