pd-l1hela细胞对人外周血活化t细胞的免疫调节作用

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1、PD-L1+HeLa细胞对人外周血活化T细胞的免疫调节作用徐光旭，徐曼，耿卫朴，陈瑜，赵涌（400016重庆，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）[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

2、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

3、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、(1006.2±30.2)pg/ml和(1070.2±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。[关键词]PD-L1；T细胞；增殖；凋亡；IFN-γ[中图法分类号]R392.3;R737.3[文献标志码]AImmunoregulatoryeffectofPD-L1+HeLacellsonactivatedhumanperipheralbloodTly

4、mphocytesXuGuangxu,XuMan,GengWeipu,ChenYu,ZhaoYong(DepartmentofPathology,MolecularMedicineandTumorResearchCenter,ChongqingMedicalUniversity,Chongqing,400016,China)[Abstract]ObjectiveTostudytheimmunoregulatoryeffectofPD-L1expressedbyhumancervicalcarcinomacelllineHeLao

5、nactivatedhumanperipheralbloodTlymphocytesinvitro.MethodsHeLacellsweretransfectedwithPD-L1+plasmid,andthenthecells(PD-L1+HeLacells)wereco-culturedwithPHA-activatedhumanperipheralbloodTlymphocytes.TheproliferationlevelofTlymphocyteswasdeterminedbyMTTassay.Theapoptosis

6、rateofTlymphocytesandtheratioofCD8+TlymphocytestoCD4+Tlymphocytes(CD8+T/CD4+T)wereanalyzedbyflowcytometry.TheIFN-γconcentrationintheculturesupernatantofTlymphocyteswasmeasuredbyELISA.ResultsTheopticaldensity(OD)ofTlymphocytesco-culturedwithPD-L1+HeLacells(0.31±0.02)w

7、assignificantlylowerthanthoseofTlymphocytesco-culturedwithHeLacellsfreeofPD-L1(PD-L1-HeLacells)(0.62±0.02)andTlymphocytesculturedalone(0.59±0.03)(P<0.01).TheapoptosisrateofTlymphocytesco-culturedwithPD-L1+HeLacells(32.7%)wassignificantlyhigherthanthoseofTlymphocytesc

8、o-culturedwithPD-L1-HeLacells(17.9%)andTlymphocytesculturedalone(18.3%)原文中文摘要给出的数据与全文中的数据不符，这里我们采用了全文实验中（2.3）的数据，还需要作者进一步确认。.TheCD8