We developed this comprehensive 28-color flow cytometry panel with the aim to measure a variety of T cell effector functions in combination with T cell differentiation markers (CCR7, CD27, CD28, CD45RO, CD95) in γδ T cells and CD4+ and CD8+ αβ T cells (Table 1). The effector functions measured in this panel include activation and co-stimulatory molecules (CD69, CD137, and CD154), cytokines (IL-2, IL-13, IL-17A, IL-21, IL-22, TNF, and IFNγ), the chemokine IL-8, cytotoxic molecules (perforin and granzyme B), and the degranulation marker CD107a. In addition, Ki67 enables the identification and analysis of recently activated T cells. To characterize regulatory T cells (Tregs ), we included CD25, CD39, and the canonical Tregs transcription factor FoxP3. We developed and optimized this panel for cryopreserved human peripheral blood mononuclear cells (PBMC) and stimulation with phorbol 12-myristate 13-acetate (PMA) and ionomycin. However, we successfully tested other types of stimulation such as staphylococcus enterotoxin B (SEB) or a mix of immunodominant peptides (CEF peptide pool) from cytomegalovirus (CMV), Epstein-Barr virus (EBV) and influenza. Published 2019. This article is a U.S. Government work and is in the public domain in the USA.

中文翻译：

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OMIP-060：30参数流式细胞仪面板，用于评估T细胞效应子功能和调节性T细胞。

我们开发了这种全面的28色流式细胞仪面板，旨在结合γδT细胞和CD4 +和CD8 +αβT中的T细胞分化标志物（CCR7，CD27，CD28，CD45RO，CD95）一起测量多种T细胞效应子功能细胞（表1）。在此面板中测量的效应子功能包括激活和共刺激分子（CD69，CD137和CD154），细胞因子（IL-2，IL-13，IL-17A，IL-21，IL-22，TNF和IFNγ） ，趋化因子IL-8，细胞毒性分子（穿孔素和颗粒酶B）和脱粒标记CD107a。此外，Ki67还可以识别和分析最近激活的T细胞。为了表征调节性T细胞（Tregs），我们包括CD25，CD39和规范性Tregs转录因子FoxP3。我们针对冷冻保存的人外周血单个核细胞（PBMC）以及用佛波12-肉豆蔻酸酯13-醋酸酯（PMA）和离子霉素刺激而开发和优化了该面板。但是，我们成功测试了其他刺激类型，例如葡萄球菌肠毒素B（SEB）或巨细胞病毒（CMV），爱泼斯坦-巴尔病毒（EBV）和流感病毒的免疫优势肽（CEF肽库）的混合物。出版于2019年。本文是美国政府的工作，在美国属于公共领域。