Interleukin-35 induces regulatory B cells that suppress autoimmune disease

Abstract

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.

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Figure 1: IL-35 induces Breg cells.
Figure 2: IL-35 preferentially induces CD5+CD19+B220lo Breg cells and an IL-35–producing Breg subpopulation (IL-35+ Breg cells).
Figure 3: rIL-35 induces expansion of Breg cells and suppresses EAU in vivo.
Figure 4: IL-35–induced Breg cells suppress uveitis.
Figure 5: IL-35 signaling is required for the suppressive function of Breg and IL-35+ Breg cells.
Figure 6: IL-35 activates STAT1 and STAT3 through an IL-35 receptor comprising IL-12Rβ2 and IL-27Rα in B cells.

Change history

  • 30 May 2014

    In the version of this article initially published online, there were several errors in the text and figures. Two authors' names were incorrect: Monika Dolinska should have been Monika B. Dolinska and Yuri V. Sergeey should have been Yuri V. Sergeev. Mann-Whitney was misspelled as Mann-Whiney in the legends of Figures 3 and 4 and in the Online Methods (Statistical analyses section). In Figure 4b, Il-10 should have been IL-10. In Figure 4i, the black key label was PMA + rIL-35 when it should have been PMA + hIL-35, and in the legend for Figure 4h,i, rIL-35 should have been hIL-35. In Figure 5e, IL-0 KO should have been IL-10 KO. In Figure 5f, IL-35 should have been rIL-35. Asterisks indicating statistical significance were missing in Figure 5d and Figure 6b. In Figure 6a, IL27Rα should have been Il27ra. In Figure 6e, on the x axis the second IL-12Rβ2 should have been IL-27Rα. In the Online Methods (Immunoprecipitation and western blot analysis section), IL-12Rb1 and IL-12Rb2 should have been IL-12Rβ1 and IL-12Rβ2. These errors have been corrected for the print, PDF and HTML versions of this article.

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Acknowledgements

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.

Author information

R.-X.W. conducted all experiments, prepared the figures and edited the manuscript. C.-R.Y. assisted with EAU experiments, FACS analysis, preparation of the figures and editing of the manuscript. I.M.D. assisted with EAU experiments, FACS analysis, protein purification, preparation of the figures and editing of the manuscript. R.M.M. performed western blot analyses and assisted with rIL-35 preparation. M.B.D. and Y.V.S. assisted with FPLC analysis. P.T.W. performed equilibrium ultracentrifugation analysis. S.-H.K. assisted with EAU experiments and fundoscopy. C.E.E. conceived, designed and supervised the project and wrote the manuscript.

Correspondence to Charles E Egwuagu.

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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

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