PD-L1^＋ HeLa细胞对人外周血活化T细胞的免疫调节作用  

Immunoregulatory effect of PD-L1^+ HeLa cells on activated human peripheral blood T lymphocytes

作　　者：徐光旭[1] 徐曼[1] 耿卫朴[1] 陈瑜[1] 赵涌[1] 

机构地区：[1]重庆医科大学基础医学院病理学教研室,分子医学与肿瘤研究中心,重庆400016

出　　处：《第三军医大学学报》2011年第16期1710-1713,共4页Journal of Third Military Medical University

基　　金：重庆市自然科学基金(CSTC2009BB5268)~~

摘　　要：目的探讨人宫颈癌细胞表达PD-L1对人外周血T细胞的免疫调节作用。方法采用PD-L1质粒转染人宫颈癌HeLa细胞株,将PD-L1+HeLa细胞与PHA刺激活化的人外周血T淋巴细胞混合培养,MTT法检测T细胞的增殖水平,流式细胞术分析T细胞凋亡率和CD8+/CD4+T细胞比例;ELISA检测PD-L1+HeLa细胞与PHA刺激活化的人外周血T淋巴细胞混合培养上清液IFN-γ的含量。结果 MTT检测显示与PD-L1+HeLa混合培养组的T细胞增殖光密度值为(0.31±0.02),明显低于与PD-L1-HeLa混合培养组的T细胞和对照组T细胞,分别为[(0.62±0.02)和(0.59±0.03),P<0.01];T细胞凋亡率明显高于与PD-L1-HeLa混合培养组和对照组T细胞,分别为(32.7%、17.9%和18.3%);CD8+/CD4+T细胞比值稍低于PD-L1-HeLa混合培养组和对照组T细胞(分别为0.86、0.91和0.89);与PD-L1+HeLa混合的T细胞培养上清液IFN-γ的含量低于与PD-L1-HeLa混合培养组和对照组T细胞,分别为[(801.1±20.5)pg/ml、(1 006.2±30.2)pg/ml和(1 070.2±113.3)pg/ml,P<0.01]。结论 PD-L1明显抑制人外周血活化T细胞增殖促进其凋亡并下调其免疫功能。Objective To study the immunoregulatory effect of PD-L1 expressed by human cervical carcinoma cell line HeLa on activated human peripheral blood T lymphocytes in vitro.Methods HeLa cells were transfected with PD-L1＋ plasmid,and then the cells（PD-L1＋ HeLa cells） were co-cultured with PHA-activated human peripheral blood T lymphocytes.The proliferation level of T lymphocytes was determined by MTT assay.The apoptosis rate of T lymphocytes and the ratio of CD8＋ T lymphocytes to CD4＋ T lymphocytes（CD8＋T/CD4＋T） were analyzed by flow cytometry.The IFN-γ concentration in the culture supernatant of T lymphocytes was measured by ELISA.Results The optical density（OD） of T lymphocytes co-cultured with PD-L1＋ HeLa cells（0.31 ±0.02） was significantly lower than those of T lymphocytes co-cultured with HeLa cells free of PD-L1（PD-L1-HeLa cells）（0.62±0.02） and T lymphocytes cultured alone（0.59±0.03）（P0.01）.The apoptosis rate of T lymphocytes co-cultured with PD-L1＋ HeLa cells（32.7%） was significantly higher than those of T lymphocytes co-cultured with PD-L1-HeLa cells（17.9%） and T lymphocytes cultured alone（18.3%）.The CD8＋T/CD4＋T ratio of T lymphocytes co-cultured with PD-L1＋ HeLa cells（0.86） was slightly lower than those of T lymphocytes co-culture with PD-L1-HeLa cells（0.91） and T lymphocytes cultured alone（0.89）.The IFN-γ concentration in the culture supernatant of T lymphocytes co-cultured with PD-L1＋ HeLa cells（801.1±20.5 pg/ml） was significantly lower than those in the T lymphocytes co-cultured with PD-L1-HeLa cells（1006.2±30.2 pg/ml） and T lymphocytes cultured alone（1070.2±113.3 pg/ml）（P0.01）.Conclusion PD-L1 significantly inhibits the proliferation in active peripheral blood T lymphocytes,promotes their apoptosis,and down-regulates their immune response.

关 键 词：PD-L1 T细胞 增殖 凋亡 IFNγ- 

分 类 号：R392.12[医药卫生—免疫学] R392.3[医药卫生—基础医学]