γδT细胞对宫颈癌细胞体外杀伤作用的探讨

In vitro Cell-Killing Action of Gamma Delta T Cells on Cervical Carcinoma Cells

目的:探讨宫颈癌组织中是否存在γδT细胞以及γδT细胞在体外抗肿瘤作用。方法:免疫组织化学法检测10例宫颈癌组织中γδT细胞的数量,固相抗体包被法体外扩增出特异性的γδT细胞后用流式细胞仪检测其纯度,乳酸脱氢酶释放实验检测γδT细胞对宫颈癌细胞株SiHa、HeLa细胞的杀伤能力。结果:免疫组化结果显示人宫颈癌组织中肿瘤浸润γδT细胞阳性染色较癌旁组织显著增强,该细胞通过固相抗体包被法体外扩增30d之后用流式细胞仪检测细胞表面TCRγδ的阳性率高达(91.2±1.2)%,乳酸脱氢酶释放实验显示体外扩增的γδT细胞能够杀伤宫颈癌细胞株SiHa、HeLa细胞,并且其杀伤效应随效应细胞/靶细胞的比例增高而增强。结论:宫颈癌组织中存在γδT细胞,该细胞在体外对宫颈癌细胞具有杀伤作用,其杀伤效应随效应细胞/靶细胞的比例增高而增强。

Objective： To investigate whether γδT cells exist in malignant tissues of cervix and the anti-tumor effect of in vitro-expanded γδT cells on the human cervical carcinoma cell lines.Methods： The numbers of γδT cells were detected in 10 cases of cervical carcinoma tissue by immunohistochemistry.γδT cells were expanded from the cervical carcinoma tissue using solid phase antibody ex vivo.Then their purity was determined by flow cytometry.Cell-killing activity of γδT cells on the human cervical carcinoma cell lines SiHa and HeLa was detected with lactic dehydrogenase release assay.Results： The results of immunohistochemisty showed that positive staining tumor-infiltrating γδT lymphocytes（γδTILs） in the human cervical carcinoma tissues were significantly increased than in the adjacent tissues.The γδTILs were expanded by solid-phase antibody method for 30 days,then the percentage of TCRγδ＋ cells in γδTILs after ex vivo expansion increased to（91.2±1.2）% detected by flow cytometry.Expanded γδTILs demonstrated marked cytotoxicity to SiHa and HeLa cells by lactic dehydrogenase release assay,and the killing effect can be enhanced with the ratio of effectors to target cells growing.Conclusion： γδT cells are found in the human cervical carcinoma tissue.And in vitro-expanded γδT cells has an anti-tumor effect in vitro.