Couillault Coline, Germain Claire, Dubois Bertrand, Kaplon Hélène
Laboratory "Targeting of the Tumor and Its Immune Microenvironment", Centre Léon Bérard, Centre de Recherche en Cancérologie de Lyon, University Claude Bernard Lyon 1, Institut National de la Santé et de la Recherche Médicale (INSERM) U1052, CNRS 5286, Lyon, France.
Cordeliers Research Center, Laboratory "Cancer, Immune Control and Escape", INSERM, UMRS 1138, Paris, France.
Methods Mol Biol. 2018;1845:205-222. doi: 10.1007/978-1-4939-8709-2_12.
Follicular helper T (Tfh) cells are major components of the humoral immune response due to their pivotal role in germinal center formation and antibody affinity maturation following B-cell isotype switching. This CD4 T-cell subtype is mainly found in the B-cell zone of secondary lymphoid organs as well as in tertiary lymphoid structures (TLS), which are highly organized structures composed of T and B cells, occasionally found at the invasive margin in the tumor microenvironment.We describe here how to perform immunofluorescence staining of tumor tissue sections and multicolor flow cytometry on tumor cell suspensions to identify and visualize these TLS-associated Tfh cells within the tumor microenvironment of various human cancers. These assays take advantage of combinations of markers and molecules involved in Tfh differentiation and function.
滤泡辅助性T（Tfh）细胞是体液免疫反应的主要组成部分，因为它们在生发中心形成以及B细胞同种型转换后的抗体亲和力成熟过程中起着关键作用。这种CD4 T细胞亚型主要存在于次级淋巴器官的B细胞区以及三级淋巴结构（TLS）中，TLS是由T细胞和B细胞组成的高度有组织的结构，偶尔出现在肿瘤微环境的浸润边缘。我们在此描述如何对肿瘤组织切片进行免疫荧光染色以及对肿瘤细胞悬液进行多色流式细胞术，以识别和可视化各种人类癌症肿瘤微环境中这些与TLS相关的Tfh细胞。这些检测利用了参与Tfh分化和功能的标志物和分子的组合。
