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IL-35-mediated induction of a potent regulatory T cell population

Abstract

Regulatory T cells (Treg cells) have a critical role in the maintenance of immunological self-tolerance. Here we show that treatment of naive human or mouse T cells with IL-35 induced a regulatory population, which we call 'iTR35 cells', that mediated suppression via IL-35 but not via the inhibitory cytokines IL-10 or transforming growth factor-β (TGF-β). We found that iTR35 cells did not express or require the transcription factor Foxp3, and were strongly suppressive and stable in vivo. Treg cells induced the generation of iTR35 cells in an IL-35- and IL-10-dependent manner in vitro and induced their generation in vivo under inflammatory conditions in intestines infected with Trichuris muris and within the tumor microenvironment (B16 melanoma and MC38 colorectal adenocarcinoma), where they contributed to the regulatory milieu. Thus, iTR35 cells constitute a key mediator of infectious tolerance and contribute to Treg cell–mediated tumor progression. Furthermore, iTR35 cells generated ex vivo might have therapeutic utility.

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Figure 1: Treatment of Tconv cells with human IL-35 confers a regulatory phenotype.
Figure 2: Treatment of Tconv cells with mouse IL-35 converts them into an IL-35-producing suppressive population.
Figure 3: Suppressive effects of iTR35 cells in vivo.
Figure 4: Stability of iTR35 cells and TGF-β–iTR cells in vivo.
Figure 5: Treg cells generate iTR35 cells in an IL-35- and IL-10-dependent manner.
Figure 6: IL-35-producing Foxp3 iTR35 cells develop in vivo.
Figure 7: The suppressive T cell milieu in the tumor microenvironment is largely due to iTR35 cells.

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Acknowledgements

We thank R. Blumberg and T. Kuo (Brigham and Women's Hospital) for Ebi3−/− mice; A. Rudensky (Memorial Sloan-Kettering Cancer Center) for Foxp3gfp mice; J. Ihle (St. Jude Children's Research Hospital; with permission from A. Rudensky) for Foxp3−/− mice; T. Geiger (St. Jude Children's Research Hospital) for Il10−/− mice; B. Triplett, D. Regan, M. Howard and M. McKenna (St. Louis Cord Blood Bank) for cord blood samples; D. Campana (St. Jude Children's Research Hospital) for the proprietary permeabilization buffer; A. Korman and M. Selby (Medarex–Bristol Myers Squibb) for MC38 colorectal adenocarcinoma cells; and S. Burns, H. Chi, R. Cross, K. Forbes, D. Green, G. Lennon, L. Jones, A. Krause, T. Moore, S. Morgan, A. Szymczak-Workman and K. Vignali for discussions and assistance. Supported by the National Institutes of Health (R01 AI39480 to D.A.A.V.; R01 AI61570 and R01 AI74878 to D.A.; and F32 AI072816 to L.W.C.), the Australian National Health and Medical Research Council Overseas Biomedical Fellowship Program (P.R.G.), the National Cancer Institute Comprehensive Cancer Center (CA21765 subaward to D.A.A.V.) and the American Lebanese Syrian Associated Charities (D.A.A.V.).

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L.W.C. designed (with help from D.A.A.V.) and did all mouse experiments, analyzed data and wrote the manuscript; V.C. did human experiments; A.L.H. (with L.W.C.) did the B16 tumor experiments; J.B. did the MC38 tumor experiments; P.R.G. infected mice with T. muris; C.G. did confocal microscopy; D.F. analyzed Affymetrix data; K.F. and S.A.B (with C.J.W.) generated and screened monoclonal antibodies to IL-35; C.J.W. coordinated the development of monoclonal anti-IL-35 and aided in figure preparation; M.L.J. generated and purified mouse Ebi3 protein for immunization and the development of monoclonal antibodies; H.-T.N. provided reagents and information; J.E.R. created and did histological analyses of Foxp3−/− mice; D.A. designed T. muris experiments and provided input on their interpretation; M.J.T. provided training for the B16 tumor model and provided input to research design and interpretation; and D.A.A.V. conceived of the research, directed the study and edited the manuscript.

Corresponding author

Correspondence to Dario A A Vignali.

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Competing interests

D.A.A.V. and L.W.C. have submitted a patent based on this work that is now pending. Also, D.A.A.V., L.W.C. and C.J.W. have also submitted a patent on IL-35 and are entitled to a share in net income generated from licensing of these patent rights for commercial development. M.L.J. is employed by Shenandoah Biotechnology and H.-T.N. is employed by R&D Systems.

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Collison, L., Chaturvedi, V., Henderson, A. et al. IL-35-mediated induction of a potent regulatory T cell population. Nat Immunol 11, 1093–1101 (2010). https://doi.org/10.1038/ni.1952

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