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The inhibitory cytokine IL-35 contributes to regulatory T-cell function

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Abstract

Regulatory T (Treg) cells are a critical sub-population of CD4+ T cells that are essential for maintaining self tolerance and preventing autoimmunity1,2, for limiting chronic inflammatory diseases, such as asthma and inflammatory bowel disease3,4, and for regulating homeostatic lymphocyte expansion5. However, they also suppress natural immune responses to parasites6 and viruses7 as well as anti-tumour immunity induced by therapeutic vaccines8. Although the manipulation of Treg function is an important goal of immunotherapy, the molecules that mediate their suppressive activity remain largely unknown. Here we demonstrate that Epstein-Barr-virus-induced gene 3 (Ebi3, which encodes IL-27β) and interleukin-12 alpha (Il12a, which encodes IL-12α/p35) are highly expressed by mouse Foxp3+ (forkhead box P3) Treg cells but not by resting or activated effector CD4+ T (Teff) cells, and that an Ebi3–IL-12α heterodimer is constitutively secreted by Treg but not Teff cells. Both Ebi3 and Il12a messenger RNA are markedly upregulated in Treg cells co-cultured with Teff cells, thereby boosting Ebi3 and IL-12α production in trans. Treg-cell restriction of this cytokine occurs because Ebi3 is a downstream target of Foxp3, a transcription factor that is required for Treg-cell development and function. Ebi3–/– and Il12a–/– Treg cells have significantly reduced regulatory activity in vitro and fail to control homeostatic proliferation and to cure inflammatory bowel disease in vivo. Because these phenotypic characteristics are distinct from those of other IL-12 family members, this novel Ebi3–IL-12α heterodimeric cytokine has been designated interleukin-35 (IL-35). Ectopic expression of IL-35 confers regulatory activity on naive T cells, whereas recombinant IL-35 suppresses T-cell proliferation. Taken together, these data identify IL-35 as a novel inhibitory cytokine that may be specifically produced by Treg cells and is required for maximal suppressive activity.

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Figure 1: Treg-restricted expression of Ebi3 and Il12a.
Figure 2: Ebi3 –/– and Il12a –/– T reg cells are functionally defective.
Figure 3: Ebi3 –/– and Il12a –/– T reg cells fail to cure IBD.
Figure 4: IL-35 suppresses T eff cell proliferation.

Change history

  • 18 January 2008

    'In the online-only extended Methods, the Il23a 5' primer, the Il23a 3' primer and the Il23a probe were corrected on 18 January 2008.

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Acknowledgements

We thank A.-M. Clark for molecular analysis; K. Forbes for colony management; J. Rogers for FACS; D. Finkelstein and T. Xu for Affymetrix GeneChip data analysis; staff at the St Jude Hartwell Center for Affymetrix GeneChip probing, oligo synthesis and DNA sequencing; staff at the St Jude ARC Histology Laboratory and Animal Husbandry Unit, and the Flow Cytometry and Cell Sorting Shared Resource facility for MACS; J. Fisher for immunoprecipitation and western blot advice; and P. Just, G. Li and D. Mitchell for Ebi3 intracellular staining and western blotting reagents. We also thank D. Green, D. Pardoll and T. Geiger for their review of the manuscript; members of the Vignali laboratory for discussions; H. von Boehmer and D. Pardoll for the 6.5 TCR transgenic mice; and A. Rudensky for the Foxp3gfp knock-in mice. This work was supported by the National Institutes of Health (D.A.A.V., R.S.B. and T.T.K.), by a Cancer Center Support CORE grant and the American Lebanese Syrian Associated Charities (ALSAC) (D.A.A.V.), by a St Jude Gephardt Postdoctoral Fellowship and an Individual NRSA (L.W.C.), and by the American Liver Foundation (T.T.K.).

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Correspondence to Dario A. A. Vignali.

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Some of us (D.A.A.V., L.W.C., C.J.W. and K.M.V.) have submitted a patent based on this work that is currently pending and licensed to a commercial entity. Furthermore, one of us (D.S.) works for a for-profit company (eBioscience).

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Collison, L., Workman, C., Kuo, T. et al. The inhibitory cytokine IL-35 contributes to regulatory T-cell function. Nature 450, 566–569 (2007). https://doi.org/10.1038/nature06306

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