Interleukin-10 (IL-10)-producing regulatory B (Breg) cells suppress autoimmune disease, and increased numbers of Breg cells prevent host defense to infection and promote tumor growth and metastasis by converting resting CD4+ T cells to regulatory T (Treg) cells. The mechanisms mediating the induction and development of Breg cells remain unclear. Here we show that IL-35 induces Breg cells and promotes their conversion to a Breg subset that produces IL-35 as well as IL-10. Treatment of mice with IL-35 conferred protection from experimental autoimmune uveitis (EAU), and mice lacking IL-35 (p35 knockout (KO) mice) or defective in IL-35 signaling (IL-12Rβ2 KO mice) produced less Breg cells endogenously or after treatment with IL-35 and developed severe uveitis. Adoptive transfer of Breg cells induced by recombinant IL-35 suppressed EAU when transferred to mice with established disease, inhibiting pathogenic T helper type 17 (TH17) and TH1 cells while promoting Treg cell expansion. In B cells, IL-35 activates STAT1 and STAT3 through the IL-35 receptor comprising the IL-12Rβ2 and IL-27Rα subunits. As IL-35 also induced the conversion of human B cells into Breg cells, these findings suggest that IL-35 may be used to induce autologous Breg and IL-35+ Breg cells and treat autoimmune and inflammatory disease.
Subscribe to Journal
Get full journal access for 1 year
only $18.75 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Edwards, J.C. et al. Efficacy of B cell–targeted therapy with rituximab in patients with rheumatoid arthritis. N. Engl. J. Med. 350, 2572–2581 (2004).
Hu, C.Y. et al. Treatment with CD20-specific antibody prevents and reverses autoimmune diabetes in mice. J. Clin. Invest. 117, 3857–3867 (2007).
Yanaba, K. et al. B-lymphocyte contributions to human autoimmune disease. Immunol. Rev. 223, 284–299 (2008).
Bouaziz, J.D. et al. Therapeutic B cell depletion impairs adaptive and autoreactive CD4+ T cell activation in mice. Proc. Natl. Acad. Sci. USA 104, 20878–20883 (2007).
Hamaguchi, Y. Molecular mechanisms of B lymphocyte depletion by CD20 immunotherapy. Nihon Rinsho Meneki Gakkai Kaishi 32, 29–34 (2009).
Pers, J.O. et al. B cell depletion and repopulation in autoimmune diseases. Clin. Rev. Allergy Immunol. 34, 50–55 (2008).
Bouaziz, J.D., Yanaba, K. & Tedder, T.F. Regulatory B cells as inhibitors of immune responses and inflammation. Immunol. Rev. 224, 201–214 (2008).
Yanaba, K. et al. A regulatory B cell subset with a unique CD1dhiCD5+ phenotype controls T cell–dependent inflammatory responses. Immunity 28, 639–650 (2008).
Mauri, C. & Bosma, A. Immune regulatory function of B cells. Annu. Rev. Immunol. 30, 221–241 (2012).
Tadmor, T., Zhang, Y., Cho, H.M., Podack, E.R. & Rosenblatt, J.D. The absence of B lymphocytes reduces the number and function of T-regulatory cells and enhances the anti-tumor response in a murine tumor model. Cancer Immunol. Immunother. 60, 609–619 (2011).
Schioppa, T. et al. B regulatory cells and the tumor-promoting actions of TNF-a during squamous carcinogenesis. Proc. Natl. Acad. Sci. USA 108, 10662–10667 (2011).
Mauri, C. & Ehrenstein, M.R. The 'short' history of regulatory B cells. Trends Immunol. 29, 34–40 (2008).
Berthelot, J.M. et al. Regulatory B cells play a key role in immune system balance. Joint Bone Spine 80, 18–22 (2013).
Yoshizaki, A. et al. Regulatory B cells control T cell autoimmunity through IL-21–dependent cognate interactions. Nature 491, 264–268 (2012).
Rafei, M. et al. A granulocyte-macrophage colony-stimulating factor and interleukin-15 fusokine induces a regulatory B cell population with immune suppressive properties. Nat. Med. 15, 1038–1045 (2009).
Collison, L.W. et al. The inhibitory cytokine IL-35 contributes to regulatory T cell function. Nature 450, 566–569 (2007).
Collison, L.W. et al. IL-35–mediated induction of a potent regulatory T cell population. Nat. Immunol. 11, 1093–1101 (2010).
Hunter, C.A. New IL-12-family members: IL-23 and IL-27, cytokines with divergent functions. Nat. Rev. Immunol. 5, 521–531 (2005).
Devergne, O., Birkenbach, M. & Kieff, E. Epstein-Barr virus–induced gene 3 and the p35 subunit of interleukin 12 form a novel heterodimeric hematopoietin. Proc. Natl. Acad. Sci. USA 94, 12041–12046 (1997).
Niedbala, W. et al. IL-35 is a novel cytokine with therapeutic effects against collagen-induced arthritis through the expansion of regulatory T cells and suppression of Th17 cells. Eur. J. Immunol. 37, 3021–3029 (2007).
Caspi, R.R. A look at autoimmunity and inflammation in the eye. J. Clin. Invest. 120, 3073–3083 (2010).
Fillatreau, S., Gray, D. & Anderton, S.M. Not always the bad guys: B cells as regulators of autoimmune pathology. Nat. Rev. Immunol. 8, 391–397 (2008).
Carter, N.A. et al. Mice lacking endogenous IL-10–producing regulatory B cells develop exacerbated disease and present with an increased frequency of Th1/Th17 but a decrease in regulatory T cells. J. Immunol. 186, 5569–5579 (2011).
Ding, Q. et al. Regulatory B cells are identified by expression of TIM-1 and can be induced through TIM-1 ligation to promote tolerance in mice. J. Clin. Invest. 121, 3645–3656 (2011).
Nussenblatt, R.B. Proctor Lecture. Experimental autoimmune uveitis: mechanisms of disease and clinical therapeutic indications. Invest. Ophthalmol. Vis. Sci. 32, 3131–3141 (1991).
Caspi, R.R. et al. A new model of autoimmune disease. Experimental autoimmune uveoretinitis induced in mice with two different retinal antigens. J. Immunol. 140, 1490–1495 (1988).
Vasconcellos, R., Carter, N.A., Rosser, E.C. & Mauri, C. IL-12p35 subunit contributes to autoimmunity by limiting IL-27–driven regulatory responses. J. Immunol. 187, 3402–3412 (2011).
Collison, L.W. et al. The composition and signaling of the IL-35 receptor are unconventional. Nat. Immunol. 13, 290–299 (2012).
Tarrant, T.K., Silver, P.B., Chan, C.C., Wiggert, B. & Caspi, R.R. Endogenous IL-12 is required for induction and expression of experimental autoimmune uveitis. J. Immunol. 161, 122–127 (1998).
Egwuagu, C.E., Charukamnoetkanok, P. & Gery, I. Thymic expression of autoantigens correlates with resistance to autoimmune disease. J. Immunol. 159, 3109–3112 (1997).
Shen, P. et al. IL-35–producing B cells are critical regulators of immunity during autoimmune and infectious diseases. Nature 507, 366–370 (2014).
Vignali, D.A. & Kuchroo, V.K. IL-12 family cytokines: immunological playmakers. Nat. Immunol. 13, 722–728 (2012).
Wang, R.X., Yu, C.R., Mahdi, R.M. & Egwuagu, C.E. Novel IL27p28/IL12p40 cytokine suppressed experimental autoimmune uveitis by inhibiting autoreactive Th1/Th17 cells and promoting expansion of regulatory T cells. J. Biol. Chem. 287, 36012–36021 (2012).
Stumhofer, J.S. et al. A role for IL-27p28 as an antagonist of gp130-mediated signaling. Nat. Immunol. 11, 1119–1126 (2010).
Nussenblatt, R.B. Bench to bedside: new approaches to the immunotherapy of uveitic disease. Int. Rev. Immunol. 21, 273–289 (2002).
Olkhanud, P.B. et al. Tumor-evoked regulatory B cells promote breast cancer metastasis by converting resting CD4 T cells to T-regulatory cells. Cancer Res. 71, 3505–3515 (2011).
Liu, X., Lee, Y.S., Yu, C.R. & Egwuagu, C.E. Loss of STAT3 in CD4+ T cells prevents development of experimental autoimmune diseases. J. Immunol. 180, 6070–6076 (2008).
Takase, H. et al. Induction of suppressors of cytokine signaling (SOCS) in the retina during experimental autoimmune uveitis (EAU): potential neuroprotective role of SOCS proteins. J. Neuroimmunol. 168, 118–127 (2005).
Oh, H.M. et al. Autoreactive memory CD4+ T lymphocytes that mediate chronic uveitis reside in the bone marrow through STAT3-dependent mechanisms. J. Immunol. 187, 3338–3346 (2011).
Fillatreau, S., Sweenie, C.H., McGeachy, M.J., Gray, D. & Anderton, S.M. B cells regulate autoimmunity by provision of IL-10. Nat. Immunol. 3, 944–950 (2002).
Paques, M. et al. Panretinal, high-resolution color photography of the mouse fundus. Invest. Ophthalmol. Vis. Sci. 48, 2769–2774 (2007).
Xu, H. et al. A clinical grading system for retinal inflammation in the chronic model of experimental autoimmune uveoretinitis using digital fundus images. Exp. Eye Res. 87, 319–326 (2008).
Chan, C.C. et al. Pathology of experimental autoimmune uveoretinitis in mice. J. Autoimmun. 3, 247–255 (1990).
Amadi-Obi, A. et al. TH17 cells contribute to uveitis and scleritis and are expanded by IL-2 and inhibited by IL-27/STAT1. Nat. Med. 13, 711–718 (2007).
Yu, C.R., Lee, Y.S., Mahdi, R.M., Surendran, N. & Egwuagu, C.E. Therapeutic targeting of STAT3 (signal transducers and activators of transcription 3) pathway inhibits experimental autoimmune uveitis. PLoS ONE 7, e29742 (2012).
Li, W., Nagineni, C.N., Hooks, J.J., Chepelinsky, A.B. & Egwuagu, C.E. Interferon-g signaling in human retinal pigment epithelial cells mediated by STAT1, ICSBP, and IRF-1 transcription factors. Invest. Ophthalmol. Vis. Sci. 40, 976–982 (1999).
Oh, H.M. et al. STAT3 protein promotes T cell survival and inhibits interleukin-2 production through up-regulation of class O Forkhead transcription factors. J. Biol. Chem. 286, 30888–30897 (2011).
The Intramural Research Programs of the NEI and NIH provided funding for this research. We thank H. Young (National Cancer Institute, NIH) and R.R. Caspi and C.-C. Chan (NEI, NIH) for critical reading of the manuscript. We also thank C.-C. Chan for expert analyses of Fundus images and histology sections. In addition, we thank A. O'Hara and C.A. Hunter (University of Pennsylvania) for providing IL-27Rα KO B cells and R. Villasmil (NEI/NIH FLOW Cytometry Core facility) for cell sorting and FACS analysis.
The authors declare no competing financial interests.
About this article
Cite this article
Wang, R., Yu, C., Dambuza, I. et al. Interleukin-35 induces regulatory B cells that suppress autoimmune disease. Nat Med 20, 633–641 (2014). https://doi.org/10.1038/nm.3554
Ageing Research Reviews (2020)
Bacterial Immunogenicity Is Critical for the Induction of Regulatory B Cells in Suppressing Inflammatory Immune Responses
Frontiers in Immunology (2020)
Molecular Immunology (2020)
Transplant International (2020)