Analysis of TGFβ1 and IL-10 Transcriptional Regulation in CTCL Cells by Chromatin Immunoprecipitation
Analysis of TGFβ1 and IL-10 Transcriptional Regulation in CTCL Cells by Chromatin Immunoprecipitation
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DOI：
10.1007/978-1-4939-0928-5_30
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发表时间：
2014-01-01
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影响因子：
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通讯作者：
Vancurova, Ivana
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文献类型：
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作者：
Chang, Tzu-Pei;Kim, Myra;Vancurova, Ivana
免疫抑制细胞因子转化生长因子 β 1 (TGF β 1) 和白细胞介素 10 (IL-10) 调节多种生物过程，包括分化、增殖、组织修复、肿瘤发生、炎症和宿主防御。 TGF beta 1 和 IL-10 的异常表达与许多类型的自身免疫和炎症性疾病以及许多类型的癌症和白血病有关。皮肤 T 细胞淋巴瘤 (CTCL) 患者的恶性 CD4+ T 细胞水平较高，这些细胞表达 IL-10 和 TGF beta 1，可抑制免疫系统并减弱抗肿瘤反应。 TGF beta 1 和 IL-10 表达的转录调节是由包括 NF kappa B 在内的多种转录因子协调的。然而，虽然 NF kappa B 对促炎和抗凋亡基因的转录调节已进行了广泛研究，但对于 NF kappa B 对免疫抑制基因的调节知之甚少。在本章中，我们描述了一种使用染色质免疫沉淀 (ChIP) 的方案，通过测量人 CTCL Hut-78 细胞中 NF kappa B p65、p50、c-Rel、Rel-B 和 p52 亚基向 TGF beta 1 和 IL-10 启动子的募集来分析 TGF beta 1 和 IL-10 的转录调控。
The immunosuppressive cytokines transforming growth factor beta 1 (TGF beta 1) and interleukin-10 (IL-10) regulate a variety of biological processes including differentiation, proliferation, tissue repair, tumorigenesis, inflammation, and host defense. Aberrant expression of TGF beta 1 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 TGF beta 1 that suppress the immune system and diminish the antitumor responses. The transcriptional regulation of TGF beta 1 and IL-10 expression is orchestrated by several transcription factors, including NF kappa B. However, while the transcriptional regulation of pro-inflammatory and anti-apoptotic genes by NF kappa B has been studied extensively, much less is known about the NF kappa B regulation of immunosuppressive genes. In this chapter, we describe a protocol that uses chromatin immunoprecipitation (ChIP) to analyze the transcriptional regulation of TGF beta 1 and IL-10 by measuring recruitment of NF kappa B p65, p50, c-Rel, Rel-B, and p52 subunits to TGF beta 1 and IL-10 promoters in human CTCL Hut-78 cells.