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目的通过构建脱嘌呤/脱嘧啶核酸内切酶1(APE1)过表达CD4请下载后查看，本文暂不支持在线获取查看简介。+CD25~+调节性T细胞细胞模型,研究在PGE2体外刺激条件下,过表达APE1对CD4~+CD25~+调节性T细胞免疫功能的影响。方法选取BALB/C小鼠,分离培养CD4~+CD25~+调节性T细胞。构建APE1过表达慢病毒载体,并转染至CD4~+CD25~+调节性T细胞,建立APE1过表达CD4~+CD25~+调节性T细胞细胞模型,作为实验组;将空载体慢病毒载体转染至等量CD4~+CD25~+调节性T细胞,作为对照组。利用外源性PGE2分别与2组细胞共培养,检测2组细胞IL-10、TGF-β、APE1 mRNA和蛋白水平,并对结果进行t检验统计分析,P<0.05为差异具有统计学意义。结果成功制备APE1基因慢病毒载体,对分离培养的CD4~+CD25~+调节性T细胞进行APE1基因慢病毒载体转染。实验组细胞TGF-β、IL-10表达量(1.17±0.42,1.02±0.12)明显低于对照组(2.69±0.21,2.25±0.31),差异均具有统计学意义;实验组APE1 mRNA的相对表达量(6.57±0.49)明显高于对照组细胞(2.24±0.23),差异具有统计学意义;APE1蛋白表达量也呈同样结果。结论在外源性PGE2刺激下,当CD4~+CD25~+调节性T细胞过表达APE1时,其分泌的细胞因子即IL-10、TGF-β下调,可能导致其免疫抑制作用的下降。
Objective To construct and purify CD4 of APE1 by purifying apurinic / apyrimidinic endonuclease 1 (APE1), and to view it later. This article is not yet available online. + CD25 ~ + regulatory T cell model to investigate the effect of APE1 overexpression on the immune function of CD4 ~ + CD25 ~ + regulatory T cells stimulated by PGE2 in vitro. Methods BALB / C mice were selected and CD4 ~ + CD25 ~ + regulatory T cells were isolated and cultured. The APE1 overexpression lentiviral vector was constructed and transfected into CD4 ~ + CD25 ~ + regulatory T cells. The APE1 overexpression CD4 ~ + CD25 ~ + regulatory T cell model was established as experimental group. Transfection to the same amount of CD4 ~ + CD25 ~ + regulatory T cells, as a control group. The levels of IL-10, TGF-β, APE1 mRNA and protein in two groups of cells were detected by exogenous PGE2 co-culture, and the results were analyzed by Student’s t test. P <0.05 was considered statistically significant. Results APE1 gene lentiviral vector was successfully prepared and APE1 gene lentiviral vector was transfected into CD4 ~ + CD25 ~ + regulatory T cells isolated and cultured. The expression of TGF-β, IL-10 in the experimental group was significantly lower than that in the control group (1.17 ± 0.42, 1.02 ± 0.12) (2.69 ± 0.21, 2.25 ± 0.31), and the difference was statistically significant. The relative expression of APE1 mRNA in the experimental group (6.57 ± 0.49) was significantly higher than that of the control group (2.24 ± 0.23), the difference was statistically significant; APE1 protein expression also showed the same result. Conclusions Under the stimulation of exogenous PGE2, when CD4 ~ + CD25 ~ + regulatory T cells overexpress APE1, the secretion of cytokines, IL-10 and TGF-β, may decrease the immunosuppressive effect.