负载乙肝核心抗原-黑色素瘤抗原-A3融合蛋白的外周血单个核细胞对人非小细胞肺癌细胞体外杀伤作用的实验  

Specific cytotoxic effect of PBMC loaded with HBVc-MAGE-A3 fusion protein on human nonsmall cell lung cancer cell in vitro

作　　者：林知 陈意 林童俊 焦顺昌[1] LIN Zhi1, CHEN Yi1, LIN Tongjun2, JIAO Shunchang1(1Department of Clinical Oncology, Chinese PLA Gneral Hospital, Beijing 100853, China; 2Dalhousie University, Halifax B3K6R8 Canad)

机构地区：[1]解放军总医院肿瘤内一科,北京100853 [2]达尔豪斯大学

出　　处：《解放军医学院学报》2018年第3期234-238,共5页Academic Journal of Chinese PLA Medical School

摘　　要：目的研究经负载乙肝核心抗原(hepatitis B core antigen,HBVc)-黑色素瘤抗原-A3(melanoma antigen gene-A3,MAGE-A3)融合蛋白的外周血单个核细胞(peripheral blood monocyte cell,PBMC)激活的细胞毒性T淋巴细胞(cytotoxic T lymphocyte,CTL)对MAGE-A3高表达人非小细胞肺癌细胞的特异性体外杀伤作用。方法从健康人外周血中分离出单个核细胞(peripheral blood monocyte cell,PBMC),使用白细胞介素-2(interleukin-2,IL-2)刺激PBMC,将负载HBVc-MAGEA3融合蛋白的PBMC与正常PBMC共培养,诱导出特异性细胞毒性T淋巴细胞,以该共培养后的PBMC作为效应细胞,以高表达MAGE-A3抗原的人非小细胞肺癌细胞株NCI-H358为靶细胞,低表达MAGE-A3抗原的细胞系HEK-293T作为对照,进行肿瘤细胞杀伤试验。结果负载HBVc-MAGE-A3融合蛋白抗原的外周血单个核细胞能够成功诱导出特异性的细胞毒性T淋巴细胞,并且对非小细胞肺癌细胞有显著的杀伤作用,其杀伤率显著高于对照组(P<0.05)。当使用MAGEA3高表达的NCI-H358细胞作为靶细胞,效靶比(E∶T)=10∶1时,HBVc-MAGE-A3组的杀伤率约为60%,HBVc组和空白对照(no template control,NTC)组的杀伤率约为35%;E∶T=20∶1时,HBVc-MAGE-A3组的杀伤率约为90%,HBVc组和NTC组的杀伤率约为45%。当使用MAGE-A3低表达的HEK-293T作为靶细胞时,三组间杀伤率差异无统计学意义。结论 MAGE-A3融合蛋白抗原负载的外周血单个核细胞可以在体外成功诱导出特异性针对非小细胞肺癌细胞的细胞毒性T淋巴细胞,并具有特异性免疫杀伤作用。Objective To evaluate the specific cytotoxic effect of cytotoxic T lymphocyte （CTL） activated by peripheral blood monocyte cell （PBMC） loaded with hepatitis B core antigen （HBVc） -melanoma antigen gene-A3 （MAGE-A3） fusion protein on human non-small cell lung cancer cell with high MAGE-A3 expression in vitro. Methods PBMCs were isolated from peripheral blood of healthy individuals and stimulated with interleukin-2 （IL-2）. The PBMCs loaded with HBVc-MAGE-A3 fusion protein were co-cultured with normal PBMCs to induce the specific cytotoxic T lymphocytes. NCI-H358 cell line with high expression of MAGE- A3 antigen was selected as the target cell and HEK-293T cell line with low expression of MAGE-A3 antigen was used as the control cell to carry out the tumor cell killing test. Results Peripheral blood mononuclear cells （PBMC） loaded with HBVc-MAGE-A3 fusion protein antigen could induce specific cytotoxic T lymphocytes （CTLs）, and it had significant cytotoxic effect on non-small cell lung cancer cells. The specific killing rate of experimental group was significantly higher than that of the control group （P 〈 0.05）. In NCI-H358 cell group, when the effector target ratio （E ： T） was 10 ： 1, the killing rate of HBVc-MAGE-A3 group was about 60%, while it was about 35% in both HBVc group and NTC group with significant difference. When E ： T = 20 ： 1, the killing rate of HBVc-MAGE-A3 group was about 90%, significantly higher than 45% in HBVc group and NTC group. And there was no significant difference between the three groups when using HEK-293T with low expression of MAGE-A3 as target cells. Conclusion Peripheral blood mononuclear cells loaded with HBVc-MAGE-A3 fusion protein antigen can successfully induce specific cytotoxic T lymphocytes （CTLs） and have specific immune killing effect on non-small cell lung cancer cells in vitro.

关 键 词：外周血单个核细胞 黑色素瘤相关抗原-A3 非小细胞肺癌 

分 类 号：R734.2[医药卫生—肿瘤]