过表达APE1对CD4^+CD25^+调节性T细胞免疫功能影响的实验研究  

An experimental investigation on Influence of APE1 overexpression on immune function of CD4^+CD25^+regulatory T cells

作　　者：王林霞[1] 汤鲁明[1] 王敏[1] 严纯雪[1] 潘国权[1] 

机构地区：[1]温州医科大学附属第二医院育英儿童医院儿童ICU,浙江温州325027

出　　处：《中华全科医学》2016年第2期182-184,共3页Chinese Journal of General Practice

基　　金：浙江省医药卫生科技计划项目(2013RCA039)

摘　　要：目的通过构建脱嘌呤/脱嘧啶核酸内切酶1(APE1)过表达CD4~^+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 research the impact of APE1 overexpression on the immune function of CD4^＋CD25^＋ Treg cells with the stimulation of PGE2 in vitro. Methods We collected BALB/C mice, isolated and cultured CD4^＋CD25^＋ Treg cells, constructed APE1 overexpression lentivirus vector and transfected into CD4^＋CD25^＋ Treg cells, establish APE1 over- expression of CD4^＋CD25^＋ Treg ceils, as the experimental group cells ;the empty vector lentiviral vectors were transfected into the same amount of CD4^＋CD25^＋ Treg cells, as the control group. In vitro, two groups were stimulated by PEG2 for 72 h, IL-10, TGF-βand APE1 mRNA levels were tested, the results were analyzed using t-test,P 〈 0.05 was considered sta- tistically significant. Results The APE1 gene lentiviral vector was constructed, APE1 gene lentiviral vector transfected the CD4^＋CD25^＋ Treg successfully. After stimulation with PGE2 to transfected APE1 overexpressed CD4^＋CD25^＋ Treg and empty vector transfected CD4^＋CD25^＋ Treg in vitro, The levels of TGF-β and IL-10 （ 1.17 ±0.42,1.02 ±0. 12 ） were significantly lower in transfected APE1 overexpressed CD4^＋CD25^＋ Treg group as compared with the other group of cells （2.69 ±0.21,2.25 ±0.31 ）, all with statistical significance ;APE1 mRNA relative expression was significantly higher in APE1 overexpressed CD4^＋CD25^＋ Treg （ 6.57 ±0.49 ） than the other group （ 2.24 ±0.23 ）, with significant difference ; APE1 protein expression also showed the same results. Conclusion PGE2 stimulation can result in the expression of APE1 of CD4^＋CD25^＋ Treg cells,and decline the immunosuppression function of the CD4^＋CD25^＋ Treg,which may be re- lated to the downregulation of IL-10 and TGF-β.

关 键 词：CD4+CD25+调节性T细胞 APE1 慢病毒载体 前列腺素E2 白细胞介素-10 转化生长因子-β 

分 类 号：R392.1[医药卫生—免疫学] R631[医药卫生—基础医学]