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Deletion of a conserved Il4 silencer impairs T helper type 1–mediated immunity

Abstract

Helper T cell differentiation involves silencing as well as activation of gene expression. We have identified a conserved silencer of the gene encoding interleukin 4 (Il4) marked by DNase I hypersensitivity (HS IV) and permissive chromatin structure in all helper T cells. Deletion of HS IV increased Il4 and Il13 transcription by naive T cells and led to T helper type 2 skewing in vitro. HS IV controlled Il4 silencing during T helper type 1 differentiation, as HS IV–deficient T helper type 1 cells that expressed interferon-γ also produced abundant interleukin 4 in vitro and in vivo. Despite mounting a vigorous interferon-γ response, HS IV–deficient mice were more susceptible to Leishmania major infection than were wild-type littermate control mice, showing a critical function for Il4 silencing in T helper type 1–mediated immunity.

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Figure 1: Cytokine gene silencing in differentiated helper T cells.
Figure 2: HS IV conservation and chromatin structure.
Figure 3: Cytokine gene expression in HS IV–deficient T cells.
Figure 4: HS IV–deficient TH1 cells fail to silence Il4.
Figure 5: Cytokine expression in HS IV–deficient TH2 cells and mast cells.
Figure 6: Increased susceptibility to infection with L. major in the absence of HS IV.
Figure 7: L. major infection of HS IV–deficient mice generates IL-4-producing TH1 cells.

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Acknowledgements

Supported by the National Institutes of Health (AI44432 and HL67664 to A.R.; AI38310 to A.H.S.; and AI20047 and AI35714 to F.W.A.), Sandler Program for Asthma Research (A.R.), Damon Runyon Cancer Research Fund (DRG-1682 to K.M.A.), Howard Hughes Medical Foundation and Ryan Foundation (S.A.), and Irvington Institute for Immunological Research and Lymphoma Research Foundation (C.H.B.).

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Correspondence to Anjana Rao.

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Supplementary information

Supplementary Fig. 1

Real-time PCR quantitation of ChIP DNA. (PDF 789 kb)

Supplementary Fig. 2

Generation of HS IV−/− mice by gene targeting. (PDF 991 kb)

Supplementary Fig. 3

IL4 feedback in HS IV−/− T cell cultures. (PDF 155 kb)

Supplementary Fig. 4

Cytokine expression by HS IV−/− BMMCs. (PDF 381 kb)

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Ansel, K., Greenwald, R., Agarwal, S. et al. Deletion of a conserved Il4 silencer impairs T helper type 1–mediated immunity. Nat Immunol 5, 1251–1259 (2004). https://doi.org/10.1038/ni1135

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