肝脏肿瘤细胞诱导T淋巴细胞PD-1表达及功能研究  

PD-1 expression of T lymphocytes induced by hepatoma cells and the function of PD-1

作　　者：陈姬[1] 吴学杰[1] 王艳[1] 余敏[1] 张乃临[1] 王贵强[1] 

机构地区：[1]北京大学第一医院,北京100034

出　　处：《传染病信息》2008年第6期361-364,共4页Infectious Disease Information

基　　金：国家自然科学基金(30771905)

摘　　要：目的研究肝脏肿瘤细胞系细胞HepG2细胞与HepG2.2.15细胞对T淋巴细胞程序性死亡分子1(PD-1)表达的影响及PD-1的作用。方法HepG2细胞、HepG2.2.15细胞分别和Jurkat细胞共同培养后,标记流式抗体,流式细胞检测仪检测Jurkat细胞PD-1的表达,ELISA检测并比较阻断组与对照组培养上清中细胞因子含量,单核细胞直接细胞毒性测定法检测并比较阻断组与对照组T淋巴细胞杀伤能力即OD值的改变。结果肝脏肿瘤细胞能诱导T淋巴细胞PD-1的表达,共培养2d时表达率分别为16.17%±2.5%(HepG2细胞)和17.43%±2.2%(HepG2.2.15细胞);阻断组培养上清中IL-2、IFN-γ、IL-10浓度分别为202.9±53.0pg/ml、88.6±4.6pg/ml和63.7±13.4pg/ml,均明显高于对照组(102.9±53.0pg/ml、39.3±4.2pg/ml和34.6±13.7pg/ml),P<0.05;阻断组Jurkat细胞对HepG2.2.15细胞杀伤的OD值为0.29±0.06,明显高于对照组(0.19±0.09),P<0.05。结论肝脏肿瘤细胞能诱导T淋巴细胞PD-1的表达;阻断PD-1/PD-L1能提高T淋巴细胞分泌细胞因子的量和杀伤能力。Objective To investigate the differences in programmed cell death-1 （PD-1） expression of T lymphocytes induced by HepG2 cells and HepG2.2.15 cells and the function of PD-1 in T lymphocytes. Methods HepG2 cells and HepG2.2.15 cells were cocultured with Jurkat cells. The levels of PD-1 were detected by flow cytometry （FCM）. The number of cytokines in culture supernatants was measured using enzyme-linked immunosorbent assay （ELISA） and compared between the blocking group and the control group. The optical density （OD） value of cytotoxic action of T lymphocytes was determined using mono-nuclear cell direct cytotoxicity （MTT） assay and compared between the two groups. Results The expression rates of PD-1 in Jurkat cells were 16.17% ± 2.5% and 17.43% ± 2.2% after the co-culture with HepG2 cells and HepG2.2.15 cells for 2 days, respectively. The levels af IL-2, IFN-γ and IL-10 in the culture supernatants in the blocking group were significantly higher than those in the control group （202.9 ± 53.0 pg/ml and 102.9 ±53.0 pg/ml, 88.6 ± 4.6 pg/ml and 39.3± 4.2 pg/ml, 63.7 ±13.4 pg/ml and 34.6 ± 13.7 pg/ml, respectively, P〈0.05）. The OD value of cytotoxic action on HepG2.2.15 cells in the blocking group was significantly higher than that in the control group （0.29 ±0.06 and 0.19± 0.09, P〈0.05）. Conclusions The PD-I expression in Jurkat cells can be induced by hepatoma cells. Blocking PD-1/PDL-1 expression can increase the number of cytokines and enhance cytotoxic action.

关 键 词：肝脏肿瘤细胞 程序性死亡分子1 诱导表达 细胞因子 

分 类 号：R735.7[医药卫生—肿瘤]