Department of Immunology, University of Toronto, Toronto, ON, Canada.
Department of Immunology, University of Toronto, Toronto, ON, Canada.
Methods Enzymol. 2020;631:305-327. doi: 10.1016/bs.mie.2019.10.001. Epub 2019 Nov 12.
Tissue-resident immune cells like innate lymphoid cells (ILCs) are regulators of local immunity and tissue homeostasis. Similar to Natural Killer (NK) cells, ILCs express germline-encoded natural cytotoxicity receptors (NCRs) that facilitate the rapid execution of effector functions. Recent advances using transgenic animal models have further uncovered the developmental, transcriptional, epigenetic, and functional differences between members of the ILC family. Isolation of ILCs, which are particularly enriched in non-lymphoid tissues, can often be challenging and time consuming. Here, we provide a simple and rapid protocol for the isolation of NK cells and ILCs from murine intestinal tissues. This protocol is suitable for Fluorescence Activated Cell Sorting (FACS) and intracellular analysis of cytokine and transcription factor expression using flow and mass cytometry.
组织驻留免疫细胞，如固有淋巴细胞 (ILC)，是局部免疫和组织稳态的调节剂。与自然杀伤 (NK) 细胞类似，ILC 表达胚系编码的天然细胞毒性受体 (NCR)，这有助于快速执行效应功能。使用转基因动物模型的最新进展进一步揭示了 ILC 家族成员之间在发育、转录、表观遗传和功能上的差异。ILC 特别富集在非淋巴组织中，因此其分离通常具有挑战性且耗时。在这里，我们提供了一种从鼠肠道组织中分离 NK 细胞和 ILC 的简单、快速的方案。该方案适用于使用流式细胞术和液质联用技术对细胞因子和转录因子表达进行荧光激活细胞分选 (FACS) 和细胞内分析。
