Skapenko Alla, Schulze-Koops Hendrik
Division of Rheumatology and Clinical Immunology, Department of Medicine IV, University of Munich, Pettenkoferstrasse 8a, 80336, Munich, Germany.
Methods Mol Biol. 2017;1585:179-187. doi: 10.1007/978-1-4939-6877-0_14.
Regulation of gene expression is essential for the differentiation of pluripotent precursor cells into specialized effector cells and, thus, for the evolution of multiorgan systems. The regulation of gene expression is controlled by a variety of "extra-genic" mechanisms, termed as epigenetic mechanisms. Obviously, alterations in such control mechanisms of gene expression may result in alterations of cellular effector functions, resulting in for example defects in cellular functions, but also, if immune cells are involved, leading to the development of immunologic disorders such as malignancies and autoimmune diseases. The analysis of epigenetic modifications is therefore pertinent not only for the understanding of the regular function of the immune system, but also for the understanding of the pathophysiology of such diseases. As interleukin-9 (IL-9) is the signature cytokine for Th9 cells, and since IL-9 plays important roles in the orchestration of a protective immune responses, the analysis of epigenetic mechanisms underlying the development of IL-9 producing effector T cells is of great interest. Here, we describe a protocol to analyze epigenetic regulation of the IL-9 gene by chromatin immunoprecipitation.
基因表达调控对于多能前体细胞分化为特化效应细胞至关重要，因此对于多器官系统的进化也至关重要。基因表达调控由多种“基因外”机制控制，这些机制被称为表观遗传机制。显然，这种基因表达控制机制的改变可能导致细胞效应功能的改变，例如导致细胞功能缺陷，而且，如果涉及免疫细胞，还会导致免疫紊乱的发生，如恶性肿瘤和自身免疫性疾病。因此，表观遗传修饰的分析不仅与理解免疫系统的正常功能相关，也与理解此类疾病的病理生理学相关。由于白细胞介素-9（IL-9）是Th9细胞的标志性细胞因子，并且由于IL-9在协调保护性免疫反应中发挥重要作用，因此分析产生IL-9的效应T细胞发育背后的表观遗传机制具有重要意义。在这里，我们描述了一种通过染色质免疫沉淀分析IL-9基因表观遗传调控的方案。
