Chapter title |
Analysis of TGFβ1 and IL-10 Transcriptional Regulation in CTCL Cells by Chromatin Immunoprecipitation.
|
---|---|
Chapter number | 30 |
Book title |
Cytokine Bioassays
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Published in |
Methods in molecular biology, January 2014
|
DOI | 10.1007/978-1-4939-0928-5_30 |
Pubmed ID | |
Book ISBNs |
978-1-4939-0927-8, 978-1-4939-0928-5
|
Authors |
Tzu-Pei Chang, Myra Kim, Ivana Vancurova, Chang TP, Kim M, Vancurova I |
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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