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Absence of signaling into CD4+ cells via C3aR and C5aR enables autoinductive TGF-β1 signaling and induction of Foxp3+ regulatory T cells

Nature Immunology volume 14, pages 162–171 (2013)Cite this article

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Abstract

Signaling through the G protein–coupled receptors for the complement fragments C3a and C5a (C3aR and C5aR, respectively) by dendritic cells and CD4+ cells provides costimulatory and survival signals to effector T cells. Here we found that when signals from C3aR and C5aR were not transduced into CD4+ cells, signaling via the kinases PI(3)Kγ, Akt and mTOR ceased, activation of the kinase PKA increased, autoinductive signaling by transforming growth factor-β1 (TGF-β1) initiated and CD4+ T cells became Foxp3+ induced regulatory T cells (iTreg cells). Endogenous TGF-β1 suppressed signaling through C3aR and C5aR by preventing the production of C3a and C5a and upregulating C5L2, an alternative receptor for C5a. The absence of signaling via C3aR and C5aR resulted in lower expression of costimulatory molecules and interleukin 6 (IL-6) and more production of IL-10. The resulting iTreg cells exerted robust suppression, had enhanced stability and suppressed ongoing autoimmune disease. Antagonism of C3aR and C5aR can also induce functional human iTreg cells.

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Figure 1: Development of iTreg cells from stimulated CD4+ cells devoid of signaling via C3aR and C5aR in the absence of DCs.
Figure 2: Absence of signaling via C3aR and C5aR in CD4+ cells dominates in the induction of iTreg cells when wild-type DCs are present.
Figure 3: The absence of signaling via C3aR and C5aR lifts restraints on PKA activation and represses PI(3)Kγ-Akt-mTOR signaling.
Figure 4: TGF-β1 and C5L2 suppress complement production and enhance the induction of iTreg cells.
Figure 5: The iTreg cells that arise when signaling via C3aR and C5aR is disabled are stable and exert robust suppressor activity in vivo.
Figure 6: The iTreg cells induced when signaling via C3aR and C5aR is disabled in CD4+ cells inhibit autoimmune disease.
Figure 7: Human iTreg cells with robust suppressor activity can be induced by antagonism of C3aR and C5aR.

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Acknowledgements

We thank J. Lambris (University of Pennsylvania) for C5aR-A synthetic peptide; C. Gerard (Harvard University) for C3ar1−/−, C3ar1−/− and Gpr77−/− mice; V.J. Kuchroo (Harvard University) for Foxp3-GFP mice; Michael Caroll (Harvard University) for C3−/− mice; B. Cobb, S.v.n. Prasad, G. Dubyak and D. Anthony Jr. for critical reading; M. Sramkoski for help with six-color flow cytometry in vivo suppression assays; H.S. Kim for immunoblot analysis; J. Letterio (Case Western Reserve University) for the inhibitor of TGF-βR1, mAb to TGF-β1 and inhibitor of Smad3 and for discussions; and P. Heeger for discussions. Supported by the US National Institutes of Health (R0123598 and EY011288 to M.E.M.; T32 HL083823-03 to M.G.S.; and AI071125 to P. Heeger) and the US Department of Defense (BC085077 to M.E.M.).

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Authors and Affiliations

  1. Institute of Pathology, Case Western Reserve University, Cleveland, Ohio, USA

    Michael G Strainic, Fengqi An, Feng Lin & M Edward Medof

  2. Laboratory of Immunology, National Institutes of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA

    Ethan M Shevach

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Contributions

M.G.S. did and analyzed the experiments and participated in their design and the writing of the manuscript; E.M.S. consulted in the design and analysis of the experiments and participated in the writing of the manuscript; F.A. did some of the experiments and prepared the knockout mice; F.L. consulted in the design and analysis of the experiments; the M.E.M. group developed original concepts and brought them to E.M.S.; all work was done in the laboratory of M.E.M. under the direction of M.E.M.; and M.E.M. designed and analyzed the experiments and wrote the manuscript.

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Correspondence to M Edward Medof.

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Strainic, M., Shevach, E., An, F. et al. Absence of signaling into CD4+ cells via C3aR and C5aR enables autoinductive TGF-β1 signaling and induction of Foxp3+ regulatory T cells. Nat Immunol 14, 162–171 (2013). https://doi.org/10.1038/ni.2499

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