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Hypersensitive site 7 of the TH2 locus control region is essential for expressing TH2 cytokine genes and for long-range intrachromosomal interactions

Nature Immunology volume 6, pages 42–48 (2005)Cite this article

Abstract

Several regulatory regions are important for the expression of genes encoding T helper type 2 (TH2) cytokines, including TH2-specific DNase I hypersensitivity sites in the TH2 cytokine locus control region. Among these sites, Rad50 hypersensitive site 7 (RHS7) shows rapid TH2-specific demethylation after antigenic stimulation. To investigate the function of RHS7 in TH2 cell differentiation, we have generated RHS7-deficient mice. CD4+ T cells and mast cells showed a notable reduction in TH2 cytokine expression in vitro and TH2 responses in vivo were considerably impaired in RHS7-deficient mice. Deletion of RHS7 did not affect the expression of a linked Rad50 gene, but it did reduce long-range intrachromosomal interactions between the locus control region and promoters of the TH2 cytokine genes. Our findings show that RHS7 is essential for the proper regulation of TH2 cytokine gene expression.

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Figure 1: Targeted deletion of RHS7.
Figure 2: Expression of Rad50 is unaffected in RHS7-deficient mice.
Figure 3: Expression of IL-4 and IL-13 is reduced in TH0 and TH2 cells from RHS7-deficient mice.
Figure 4: Expression of IL-4 and IL-13 is reduced in BMMCs from RHS7-deficient mice.
Figure 5: Reduced expression of TH2 cytokines in RHS7-deficient mice on immunization with KLH.
Figure 6: Impaired interaction of RHS4 and RHS6 with the Il4 promoter in RHS7-deficient CD4+ T cells.
Figure 7: RHS7 deletion does not affect the formation of active chromatin in the LCR or the whole locus.

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Acknowledgements

We thank L. Evangelisti, C. Hughes and J. Stein for assisting with the generation of RHS7-deficient mice; P.E. Fields, S.T. Kim and E. Eynon for discussion; and F. Manzo for help with manuscript preparation. Supported by the National Institutes of Health (HL56389 to R.A.F.), Cancer Research Institute (C.S.) and Howard Hughes Medical Institute (R.A.F.). R.A.F. is an investigator of the Howard Hughes Medical Institute.

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  1. Section of Immunobiology, Yale School of Medicine, New Haven, 06520, Connecticut, USA

    Gap Ryol Lee, Charalampos G Spilianakis & Richard A Flavell

  2. Howard Hughes Medical Institute, Yale School of Medicine, New Haven, 06520, Connecticut, USA

    Richard A Flavell

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Correspondence to Richard A Flavell.

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Lee, G., Spilianakis, C. & Flavell, R. Hypersensitive site 7 of the TH2 locus control region is essential for expressing TH2 cytokine genes and for long-range intrachromosomal interactions. Nat Immunol 6, 42–48 (2005). https://doi.org/10.1038/ni1148

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