Isolation, Characterization, and Culture of Intestinal Intraepithelial Lymphocytes
Isolation, Characterization, and Culture of Intestinal Intraepithelial Lymphocytes
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DOI：
10.1007/978-1-0716-0338-3_13
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发表时间：
2020-01-01
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影响因子：
--
通讯作者：
Swamy, Mahima
中科院分区：
文献类型：
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作者：
James, Olivia Jane;Vandereyken, Maud;Swamy, Mahima
肠上皮内淋巴细胞（IEL）包括不同的先天样T细胞和记忆T细胞群，它们共同形成体内最大的T细胞区室之一。IEL位于肠上皮内，是肠道中第一个与肠道不断接触的食物、微生物群和病原体相互作用的免疫细胞。IEL可以对外界损伤做出快速反应以保护小肠上皮，但也被认为是维持肠道内稳态的重要调节细胞。然而，IEL激活的机制及其在上皮内的相互作用在很大程度上仍然是难以捉摸的。事实上，即使在肠道免疫学研究中，IEL也没有得到普遍评估，可能是因为它们被认为难以分离和研究。在本方案中，我们提出了一种从小鼠小肠中分离IEL的简化方法，并为不同IEL亚群的流式细胞分析提供了代表性数据。我们还概述了培养IEL的两个程序，可以进行功能研究和与上皮细胞共培养。这些策略应该使研究这个大而神秘的T细胞区室更容易获得，并打开对肠道内稳态机制和组织相关免疫的理解。
Intestinal intraepithelial lymphocytes (IEL) comprise distinct groups of innate-like and memory T cells that collectively form one of the largest T cell compartments in the body. IEL are located within the intestinal epithelium and are the first immune cells in the gut to interact with the food, microbiota, and pathogens that the gut is continually exposed to. IEL can respond rapidly to external insults to protect the small intestinal epithelium but are also considered regulatory cells that are important to maintain the homeostasis of the gut. However, the mechanisms of IEL activation and their interactions within the epithelium remain largely elusive. Indeed, IEL are not commonly evaluated even in studies of gut immunology, potentially because they are perceived as being difficult to isolate and study. In this protocol, we present a simplified method to isolate IEL from the murine small intestine and provide representative data for flow cytometric analyses of the different IEL subsets. We also outline two procedures for culturing IEL, which can permit functional studies and coculture with epithelial cells. These strategies should make studies of this large but enigmatic T cell compartment more accessible and open up understanding of homeostatic mechanisms in the intestine, and tissue-associated immunity.