Cardiovascular Research

NAD(+) regulates Treg cell fate and promotes allograft survival via a systemic IL-10 production that is CD4(+) CD25(+) Foxp3(+)

Authors: Abdallah Elkhal|||Hector Rodriguez Cetina Biefer|||Timm Heinbokel|||Hirofumi Uehara|||Markus Quante|||Midas Seyda|||Jeroen M Schuitenmaker|||Felix Krenzien|||Virginia Camacho|||Miguel A de la Fuente|||Ionita Ghiran|||Stefan G Tullius

Affiliations: + Expand

Abstract

CD4(+) CD25(+) Foxp3(+) Tregs have been shown to play a central role in immune homeostasis while preventing from fatal inflammatory responses, while Th17 cells have traditionally been recognized as pro-inflammatory mediators implicated in a myriad of diseases. Studies have shown the potential of Tregs to convert into Th17 cells, and Th17 cells into Tregs. Increasing evidence have pointed out CD25 as a key molecule during this transdifferentiation process, however molecules that allow such development remain unknown. Here, we investigated the impact of NAD(+) on the fate of CD4(+) CD25(+) Foxp3(+) Tregs in-depth, dissected their transcriptional signature profile and explored mechanisms underlying their conversion into IL-17A producing cells. Our results demonstrate that NAD(+) promotes Treg conversion into Th17 cells in vitro and in vivo via CD25 cell surface marker. Despite the reduced number of Tregs, known to promote homeostasis, and an increased number of pro-inflammatory Th17 cells, NAD(+) was able to promote an impressive allograft survival through a robust systemic IL-10 production that was CD4(+) CD25(+) Foxp3(+) independent. Collectively, our study unravels a novel immunoregulatory mechanism of NAD(+) that regulates Tregs fate while promoting allograft survival that may have clinical applications in alloimmunity and in a wide spectrum of inflammatory conditions.

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Chemical List

CD4 Antigens|||Forkhead Transcription Factors|||Foxp3 protein, mouse|||Interleukin-2 Receptor alpha Subunit|||NAD|||Interleukin-10

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Reference List

Fontenot J. D., Gavin M. A. & Rudensky A. Y. Foxp3 programs the development and function of CD4 CD25 regulatory T cells. Nat Immunol 4, 330–336, doi: 10.1038/ni904ni904 (2003).|||Hori S., Nomura T. & Sakaguchi S. Control of regulatory T cell development by the transcription factor Foxp3. Science 299, 1057–1061, doi: 10.1126/science (2003).|||Josefowicz S. Z., Lu L. F. & Rudensky A. Y. Regulatory T cells: mechanisms of differentiation and function. Annu Rev Immunol 30, 531–564, doi: 10.1146/annurev.immunol.25.022106.141623 (2012).|||Sakaguchi S., Sakaguchi N., Asano M., Itoh M. & Toda M. Immunologic self-tolerance maintained by activated T cells expressing IL-2 receptor alpha-chains (CD25). Breakdown of a single mechanism of self-tolerance causes various autoimmune diseases. J Immunol 155, 1151–1164 (1995).|||Zhou X. Instability of the transcription factor Foxp3 leads to the generation of pathogenic memory T cells . Nat Immunol 10, 1000–1007, doi: 10.1038/ni.1774 (2009).|||Lee Y. K., Mukasa R., Hatton R. D. & Weaver C. T. Developmental plasticity of Th17 and Treg cells. Curr Opin Immunol 21, 274–280, doi: 10.1016/j.coi.2009.05.021 (2009).|||Komatsu N. Heterogeneity of natural Foxp3 T cells: a committed regulatory T-cell lineage and an uncommitted minor population retaining plasticity. Proc Natl Acad Sci USA 106, 1903–1908, doi: 10.1073/pnas.0811556106 (2009).|||Miyao T. Plasticity of Foxp3(+) T cells reflects promiscuous Foxp3 expression in conventional T cells but not reprogramming of regulatory T cells. Immunity 36, 262–275, doi: 10.1016/j.immuni.2011.12.012 (2012).|||Gagliani N. Coexpression of CD49b and LAG-3 identifies human and mouse T regulatory type 1 cells. Nat Med 19, 739–746, doi: 10.1038/nm.3179 (2013).|||Voo K. S. Identification of IL-17-producing FOXP3 regulatory T cells in humans. Proc Natl Acad Sci USA 106, 4793–4798, doi: 10.1073/pnas.0900408106 (2009).|||Komatsu N. Pathogenic conversion of Foxp3 T cells into TH17 cells in autoimmune arthritis. Nat Med 20, 62–68, doi: 10.1038/nm.3432 (2014).|||Gagliani N. Th17 cells transdifferentiate into regulatory T cells during resolution of inflammation. Nature. doi: 10.1038/nature14452 (2015).|||Ayyoub M. Human memory FOXP3 Tregs secrete IL-17 and constitutively express the T(H)17 lineage-specific transcription factor RORgamma t. Proc Natl Acad Sci USA 106, 8635–8640, doi: 10.1073/pnas.0900621106 (2009).|||Beriou G. IL-17-producing human peripheral regulatory T cells retain suppressive function. Blood 113, 4240–4249, doi: 10.1182/blood-2008-10-183251 (2009).|||Kryczek I. IL-17 regulatory T cells in the microenvironments of chronic inflammation and cancer. J Immunol 186, 4388–4395, doi: 10.4049/jimmunol.1003251 (2011).|||Teege S. Tuning IL-2 signaling by ADP-ribosylation of CD25. Sci Rep 5, 8959, doi: 10.1038/srep08959 (2015).|||Hawiger D. & Flavell R. A. Regulatory T cells that become autoaggressive. Nat Immunol 10, 938–939, doi: 10.1038/ni0909-938 (2009).|||Zhu J., Yamane H. & Paul W. E. Differentiation of effector CD4 T cell populations (*). Annu Rev Immunol 28, 445–489, doi: 10.1146/annurev-immunol-030409-101212 (2010).|||Yang X. O. Molecular antagonism and plasticity of regulatory and inflammatory T cell programs. Immunity 29, 44–56, doi: 10.1016/j.immuni.2008.05.007 (2008).|||Xu L., Kitani A., Fuss I. & Strober W. Cutting edge: regulatory T cells induce CD4 CD25-Foxp3- T cells or are self-induced to become Th17 cells in the absence of exogenous TGF-beta. J Immunol 178, 6725–6729, doi: 10.4049/ jimmunol.178.11.6725 (2007).|||Martinez G. J., Nurieva R. I., Yang X. O. & Dong C. Regulation and function of proinflammatory TH17 cells. Ann N Y Acad Sci 1143, 188–211, doi: 10.1196/annals.1443.021 (2008).|||Laurence A. Interleukin-2 signaling via STAT5 constrains T helper 17 cell generation. Immunity 26, 371–381, doi: 10.1016/j.immuni.2007.02.009 (2007).|||Liao W., Lin J. X., Wang L., Li P. & Leonard W. J. Modulation of cytokine receptors by IL-2 broadly regulates differentiation into helper T cell lineages. Nat Immunol 12, 551–559, doi: 10.1038/ni.2030 (2011).|||Tullius S. G. NAD(+) protects against EAE by regulating CD4(+) T-cell differentiation. Nat Commun 5, 5101, doi: 10.1038/ncomms6101 (2014).|||Langrish C. L. IL-23 drives a pathogenic T cell population that induces autoimmune inflammation. J Exp Med 201, 233–240, doi: 10.1084/jem.20041257 (2005).|||Zhu J. & Paul W. E. Peripheral CD4 T-cell differentiation regulated by networks of cytokines and transcription factors. Immunol Rev 238, 247–262, doi: 10.1111/j.1600-065X.2010.00951.x (2010).|||Laurence A. STAT3 transcription factor promotes instability of nTreg cells and limits generation of iTreg cells during acute murine graft-versus-host disease. Immunity 37, 209–222, doi: 10.1016/j.immuni.2012.05.027 (2012).|||Junger W. G. Immune cell regulation by autocrine purinergic signalling. Nat Rev Immunol 11, 201–212, doi: 10.1038/nri2938 (2011).|||Casati A. Cell-autonomous regulation of hematopoietic stem cell cycling activity by ATP. Cell Death Differ 18, 396–404, doi: 10.1038/cdd.2010.107 (2011).|||Hubert S. Extracellular NAD shapes the Foxp3 regulatory T cell compartment through the ART2-P2X7 pathway. J Exp Med 207, 2561–2568, doi: 10.1084/jem.20091154 (2010).|||Schwarz N. Activation of the P2X7 ion channel by soluble and covalently bound ligands. Purinergic Signal 5, 139–149, doi: 10.1007/s11302-009-9135-5 (2009).|||Seman M. NAD-induced T cell death: ADP-ribosylation of cell surface proteins by ART2 activates the cytolytic P2X7 purinoceptor. Immunity 19, 571–582, doi: 10.1016/S1074-7613(03)00266-8 (2003).|||Donnelly-Roberts D. L. [3H]A-804598 ([3H]2-cyano-1-[(1S)-1-phenylethyl]-3-quinolin-5-ylguanidine) is a novel, potent, and selective antagonist radioligand for P2X7 receptors. Neuropharmacology 56, 223–229, doi: 10.1016/j.neuropharm.2008.06.012 (2009).|||Able S. L. Receptor localization, native tissue binding and occupancy for centrally penetrant P2X7 antagonists in the rat. Br J Pharmacol 162, 405–414, doi: 10.1111/j.1476-5381.2010.01025.x (2011).|||Boyer J. L., Adams M., Ravi R. G., Jacobson K. A. & Harden T. K. 2-Chloro N(6)-methyl-(N)-methanocarba-2′-deoxyadenosine-3′,5′-bisphosphate is a selective high affinity P2Y(1) receptor antagonist. Br J Pharmacol 135, 2004–2010, doi: 10.1038/sj.bjp.0704673 (2002).|||Hanidziar D. & Koulmanda M. Inflammation and the balance of Treg and Th17 cells in transplant rejection and tolerance. Curr Opin Organ Transplant 15, 411–415, doi: 10.1097/MOT.0b013e32833b7929 (2010).|||Heidt S., Segundo D. S., Chadha R. & Wood K. J. The impact of Th17 cells on transplant rejection and the induction of tolerance. Curr Opin Organ Transplant 15, 456–461, doi: 10.1097/MOT.0b013e32833b9bfb (2010).|||Asadullah K., Sterry W. & Volk H. D. Interleukin-10 therapy–review of a new approach. Pharmacol Rev 55, 241–269, doi: 10.1124/pr.55.2.4 (2003).|||Sakaguchi S. Naturally arising Foxp3-expressing CD25 CD4 regulatory T cells in immunological tolerance to self and non-self. Nat Immunol 6, 345–352, doi: 10.1038/ni1178 (2005).|||Elkhal A. CD1d restricted natural killer T cells are not required for allergic skin inflammation. J Allergy Clin Immunol 118, 1363–1368, doi: 10.1016/j.jaci.2006.08.010 (2006).|||Woehrle T. Pannexin-1 hemichannel-mediated ATP release together with P2X1 and P2X4 receptors regulate T-cell activation at the immune synapse. Blood 116, 3475–3484, doi: 10.1182/blood-2010-04-277707 (2010).