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A T-cell-specific CD154 transcriptional enhancer located just upstream of the promoter

Genes & Immunity volume 9, pages 640–649 (2008)Cite this article

Abstract

CD154 (CD40-ligand) is a critical immune regulator. CD154 expression is tightly regulated and largely restricted to activated CD4 T cells. Using DNase I hypersensitivity site (HSS) mapping, we identified two novel HSS mapping to the human CD154 promoter element and just upstream. Both HSS were activation independent and CD4 T-cell specific. Approximately 350 bp of DNA sequence flanking the upstream HSS site was highly conserved between mouse and man, and was rich in binding sites for GATA and NFAT proteins. Gel shift and chromatin immunoprecipitation assays demonstrated both NFAT1 and the Th2 factor, GATA-3, bound this enhancer element in vitro and in vivo, respectively. A PstI/XbaI 345 bp fragment of this region acted as a transcriptional enhancer of the CD154 promoter in primary human CD4 T cells. Overexpression of repressor of GATA and a dominant negative GATA-3 protein independently inhibited transcription, whereas overexpression of wild-type GATA-3 enhanced transcriptional activity, by this element in primary CD4 T cells. Moreover, more interleukin-4-producing CD4 T cells expressed CD154 following activation than interferon-γ-producing CD4 T cells. Thus, we identified a novel T-cell-specific, GATA-3 responsive, CD154 transcriptional enhancer, which may contribute to increased propensity of Th2 cells to express CD154.

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Acknowledgements

We thank Dr Kate Sullivan, Dr Gerd Blobel, Dr Ginny Shapiro, Dr Seth Lederman and Dr Mariuz Wasik for the generous gifts of reagents and cell lines as detailed in the Materials and methods. We also thank Dr Terri Finkel and Dr Gerd Blobel for critical reviews of the article. This work was supported, in part, by grants from the Arthritis Foundation, the Arthritis National Research Foundation, the Dorough Lupus Foundation, the Kahn Foundation for Lupus Research, the American College of Rheumatology Research and Education Foundation and the National Institutes of Health (R01-AR48257, R21-AR49335, P30-HH2815, M01-RR240; RQC).

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Author notes

  1. M Brunner

    Present address: 4Current address: Medical Diagnostic Laboratories, Hamilton, NJ 08690, USA.,

Authors and Affiliations

  1. Division of Rheumatology, Children's Hospital of Philadelphia, Philadelphia, PA, USA

    M Brunner, M Zhang, A Genin & R Q Cron

  2. Division of Rheumatology, Immunology and Allergy, Brigham and Women's Hospital, Boston, MA, USA

    I-C Ho

  3. Department of Pediatrics, University of Pennsylvania School of Medicine, Philadelphia, PA, USA

    R Q Cron

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  1. M Brunner
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Correspondence to R Q Cron.

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Brunner, M., Zhang, M., Genin, A. et al. A T-cell-specific CD154 transcriptional enhancer located just upstream of the promoter. Genes Immun 9, 640–649 (2008). https://doi.org/10.1038/gene.2008.67

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