Department of Microbiology and Immunology, University of Illinois-College of Medicine, Chicago, IL, USA.
Core Genomics Facility, University of Illinois at Chicago, Chicago, IL, USA.
Methods Mol Biol. 2021;2248:63-71. doi: 10.1007/978-1-0716-1130-2_4.
TNF receptor superfamily comprises many T-cell costimulatory receptors, including TNFRSF1, TNFRSF2, TNFRSF4 (OX40), TNFRSF9 (4-1BB), TNFRSF18 (GITR), and TNFRSF7 (CD27). Signaling through these costimulatory stimulatory receptors can promote conventional T-cell (Tconv) proliferation, and effector functions in an antigen-dependent manner. Thus, agonistic antibodies and ligands for OX40, 4-1BB, GITR, and CD27 have been tested for inducing T-cell-mediated antitumor responses in several cancers. However, recently emerging reports show critical role for TNFR signaling in regulatory T-cell (Treg) differentiation and expansion, which might suppress effector T-cell proliferation and functions. Here, we show preferential over expression of TNFR2, OX40, 4-1BB, and GITR in Treg cells over Tconv cells, and the ability of OX40L and GITRL to induce selective proliferation of Treg cells, but not Tconv cells, in an antigen-independent manner. We describe the standard protocols used for Affymetrix gene expression profiling, T-cell isolation, and Cell Trace Violet-based cell proliferation assay.
TNF 受体超家族包含许多 T 细胞共刺激受体，包括 TNFRSF1、TNFRSF2、TNFRSF4（OX40）、TNFRSF9（4-1BB）、TNFRSF18（GITR）和 TNFRSF7（CD27）。这些共刺激刺激受体的信号传导可以以抗原依赖性方式促进常规 T 细胞（Tconv）的增殖和效应功能。因此，已在几种癌症中测试了 OX40、4-1BB、GITR 和 CD27 的激动性抗体和配体，以诱导 T 细胞介导的抗肿瘤反应。然而，最近的报告显示，TNFR 信号在调节性 T 细胞（Treg）分化和扩增中起关键作用，这可能会抑制效应 T 细胞的增殖和功能。在这里，我们显示 TNFR2、OX40、4-1BB 和 GITR 在 Treg 细胞中的表达水平高于 Tconv 细胞，并且 OX40L 和 GITRL 能够以抗原非依赖性方式诱导 Treg 细胞而非 Tconv 细胞的选择性增殖。我们描述了用于 Affymetrix 基因表达谱分析、T 细胞分离和 CellTrace Violet 基于细胞增殖测定的标准方案。
