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Analysis of TGFβ1 and IL-10 Transcriptional Regulation in CTCL Cells by Chromatin Immunoprecipitation

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Book cover Cytokine Bioassays

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1172))

Abstract

The immunosuppressive cytokines transforming growth factor β1 (TGFβ1) and interleukin-10 (IL-10) regulate a variety of biological processes including differentiation, proliferation, tissue repair, tumorigenesis, inflammation, and host defense. Aberrant expression of TGFβ1 and IL-10 has been associated with many types of autoimmune and inflammatory disorders, as well as with many types of cancer and leukemia. Patients with cutaneous T cell lymphoma (CTCL) have high levels of malignant CD4+ T cells expressing IL-10 and TGFβ1 that suppress the immune system and diminish the antitumor responses. The transcriptional regulation of TGFβ1 and IL-10 expression is orchestrated by several transcription factors, including NFκB. However, while the transcriptional regulation of pro-inflammatory and anti-apoptotic genes by NFκB has been studied extensively, much less is known about the NFκB regulation of immunosuppressive genes. In this chapter, we describe a protocol that uses chromatin immunoprecipitation (ChIP) to analyze the transcriptional regulation of TGFβ1 and IL-10 by measuring recruitment of NFκB p65, p50, c-Rel, Rel-B, and p52 subunits to TGFβ1 and IL-10 promoters in human CTCL Hut-78 cells.

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Acknowledgements

This work was supported by NIH grant CA173452 to I. Vancurova.

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Authors and Affiliations

  1. Department of Biology, St. John’s University, 8000 Utopia Parkway, Queens, NY, 11439, USA

    Tzu-Pei Chang, Myra Kim & Ivana Vancurova

Authors
  1. Tzu-Pei Chang
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  2. Myra Kim
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  3. Ivana Vancurova
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Corresponding author

Correspondence to Ivana Vancurova .

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Editors and Affiliations

  1. Department of Biological Sciences, St. John's University, New York, New York, USA

    Ivana Vancurova

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Chang, TP., Kim, M., Vancurova, I. (2014). Analysis of TGFβ1 and IL-10 Transcriptional Regulation in CTCL Cells by Chromatin Immunoprecipitation. In: Vancurova, I. (eds) Cytokine Bioassays. Methods in Molecular Biology, vol 1172. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0928-5_30

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  • DOI: https://doi.org/10.1007/978-1-4939-0928-5_30

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0927-8

  • Online ISBN: 978-1-4939-0928-5

  • eBook Packages: Springer Protocols

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