Department of Internal Medicine- Centre of Excellence for Biomedical Research, University of Genoa- IRCCS Ospedale Policlinico San Martino, Genoa, Italy.
Centre of Excellence for Biomedical Research, University of Genoa, Genoa, Italy.
Methods Mol Biol. 2021;2285:35-47. doi: 10.1007/978-1-0716-1311-5_3.
The development of T helper (Th) cell subsets requires activated T cells that respond to a polarizing cytokine environment, resulting in the activation and expression of specific transcription factors. The subset-specific transcription factors are located either in the cytoplasm or in the nucleus, which determine the functional profile of Th populations, inducing the production of specific effector cytokines and functions. Flow cytometry analysis of transcription factors has become very common not only in research but also in immunologic follow-up protocols of patients recruited in clinical trials (as evaluation of CD4+CD25+ FOXP3+ T regulatory cells). Here, we propose and describe one-step protocols to evaluate the expression of transcription factors in mouse and human CD4+ lymphocytes, focusing the critical points of this cytometric approach.
辅助性 T 细胞（Th）亚群的发育需要对极化细胞因子环境有反应的活化 T 细胞，从而导致特定转录因子的激活和表达。亚群特异性转录因子位于细胞质或细胞核中，决定 Th 细胞群体的功能特征，诱导特定效应细胞因子和功能的产生。不仅在研究中，而且在临床试验中招募的患者的免疫随访方案中（作为评估 CD4+CD25+FOXP3+调节性 T 细胞），流式细胞术分析转录因子也变得非常普遍。在这里，我们提出并描述了一步法方案，用于评估小鼠和人 CD4+淋巴细胞中转录因子的表达，重点介绍了这种细胞测定方法的关键点。
