Molecular Neuroimmunology Unit, IRCCS Fondazione Santa Lucia, Rome, Italy.
Department of Biology and Biotechnology Charles Darwin, Sapienza University, Rome, Italy.
Methods Mol Biol. 2021;2285:27-34. doi: 10.1007/978-1-0716-1311-5_2.
T helper (Th) cells are involved in various physiopathological systems, including response to infections, vaccination, cancer, and autoimmunity. The isolation of viable human Th cells is a procedure that allows a broad study of both phenotypical and functional features of each Th subset, and thus, it is necessary to study these cells in different contexts. In particular, the purification of human memory Th cells from peripheral blood is preparatory for further complex experiments on these cells, such as global transcriptional analysis, coculture assays, silencing experiments, and drug assays.Here, we describe the method for the identification and isolation of pure memory human Th1, Th2, Th17, Th1/17, and T regulatory cells, derived from peripheral blood mononuclear cells. Moreover, we show the purity of each purified Th subset, verified by the analysis of specific transcription factors.
辅助性 T 细胞（Th 细胞）参与多种生理病理系统，包括感染、疫苗接种、癌症和自身免疫的反应。活的人类 Th 细胞的分离是一种能够广泛研究每个 Th 亚群的表型和功能特征的方法，因此，有必要在不同的环境中研究这些细胞。特别是，从外周血中纯化人类记忆 Th 细胞是进一步对这些细胞进行复杂实验的准备，如全转录组分析、共培养测定、沉默实验和药物测定。在这里，我们描述了从外周血单个核细胞中鉴定和分离纯记忆人类 Th1、Th2、Th17、Th1/17 和 T 调节细胞的方法。此外，我们通过分析特定的转录因子来显示每个纯化的 Th 亚群的纯度。
