Biram Adi, Shulman Ziv
Department of Immunology, Weizmann Institute of Science, Rehovot, Israel.
Bio Protoc. 2020 Mar 20;10(6):e3562. doi: 10.21769/BioProtoc.3562.
T follicular helper (Tfh) cells regulate B cell selection for entry into the germinal center (GC) reaction or for differentiation into antibody forming cells. This process takes place at the border between the T and B zones in lymphoid organs and involves physical contacts between T and B cells. During these interactions, T cells endow the B cells with selection signals that promote GC seeding or plasmablast differentiation based on their B cell receptor affinity. In Peyer's patches (PPs), T cells promote B cell colonization of the subepithelial dome (SED) without effective affinity-based clonal selection. To specifically characterize the T cell population that resides within the SED niche, we performed ex vivo photoactivation of the SED compartment followed by flow cytometry analysis of the labeled cells, as described in this protocol. This technique integrates both spatial and cellular information in studies of immunological niches and can be adapted to various experimental systems.
滤泡辅助性T（Tfh）细胞调节B细胞选择，使其进入生发中心（GC）反应或分化为抗体形成细胞。这一过程发生在淋巴器官中T区和B区的交界处，涉及T细胞与B细胞之间的物理接触。在这些相互作用过程中，T细胞根据B细胞受体亲和力赋予B细胞选择信号，促进GC定植或浆母细胞分化。在派尔集合淋巴结（PP）中，T细胞促进B细胞在黏膜下穹窿（SED）定植，而无需基于亲和力的有效克隆选择。为了特异性地表征位于SED生态位内的T细胞群体，我们按照本方案所述，对SED区室进行了体外光激活，随后对标记细胞进行了流式细胞术分析。该技术在免疫生态位研究中整合了空间和细胞信息，并且可适用于各种实验系统。
