γδT细胞的高效扩增及其对消化道肿瘤细胞的体外杀伤研究  

Cytotoxic effects of human γδT cells stimulated by zoledronate on digestive system carcinoma cells

作　　者：苏丹[1] 张纯慧[1] 王东亮[1] 张艳桥[1] SU Dan;ZHANG Chun-hui;WANG Dong-liang;ZHANG Yan-qiao(Department of Internal Medicine,Cancer Hospital of Harbin Medical University,Harbin 150081,China)

机构地区：[1]哈尔滨医科大学附属肿瘤医院消化内科二病房,黑龙江哈尔滨150081

出　　处：《哈尔滨医科大学学报》2018年第5期430-433,共4页Journal of Harbin Medical University

摘　　要：目的探讨人外周血扩增的γδT细胞体外扩增能力及其对消化道肿瘤细胞的杀伤作用。方法用含唑来膦酸和rh IL-2的RPMI 1640培养基培养正常人外周血单个核细胞(PBMC),流式细胞术检测活化前后γδT细胞的百分比,乳酸脱氢酶释放法测定经不同浓度唑来膦酸(5μmol/L、10μmol/L、15μmol/L、20μmol/L)体外扩增后的γδT细胞与消化道肿瘤细胞株(HCT-116、HCCC-9810、LOVO、GBCSD)按不同效靶比(3∶1,10∶1,30∶1)孵育后进行杀伤活性测定。结果 (1)流式细胞术结果显示:不同浓度的唑来膦酸(5μmol/L、10μmol/L、15μmol/L、20μmol/L)均可促进γδT细胞增殖,在5μmol/L浓度的唑来膦酸组对γδT细胞增殖促进作用最明显(P <0. 05)。(2)γδT流式测定结果显示:PBMC经唑来膦酸体外诱导后γδT细胞比例显著升高(96. 15%±1. 4849%,P=0. 0001 <0. 05)。(3)杀伤检测结果显示:γδT细胞对消化道肿瘤有杀伤活性,在效靶比为30∶1时对胆管癌细胞(HCCC-9810)及肠癌细胞(HCT116)的杀伤率分别为21. 90%±6. 6468%和30. 15%±8. 2731%。结论人末梢血PBMC经Zol+IL-2活化后可获得大量纯度较高的γδT细胞,在体外对消化道肿瘤有杀伤作用。γδT细胞将成为肿瘤细胞免疫治疗中又一重要的免疫效应细胞。Objective To investigate the cytotoxic effects of human peripheral blood γδT cells on the tumor cell lines of digestive system. Methods The γδT cells in peripheral blood of healthy volunteers were expanded using interleukin-2( IL-2) and Zol in RPMI 1640 medium.The purity of γδT cells was analyzed by flow cytometry. The cytotoxic activity of the purifiedγδT cells against human carcinoma cells were tested in different effector-to-target ratios. Results(1)The flow results showed that different concentrations of zoledronic acid( 5 μmol/L,10 μmol/L,15 μmol/L,20 μmol/L) can promote the proliferation of γδT cells,and the 5μmol/L of zoledronic group was significantly improved( P < 0. 05);(2)γδT cell stream measurement results showed that the purity of γδT cell was significantly higher( 96. 15% ±1. 4849%,P = 0. 0001 < 0. 05);(3) LDH results showed that γδT cell killed gastrointestinal tumors,at the efficiency target ratio of 30 ∶ 1 for cholangiocarcinoma cells( HCCC-9810) andcolon cancer cells( HCT116) the killing rates were( 21. 90% ± 6. 6468%) and( 30. 15% ±8. 2731%) respectively. Conclusion The γδT cells expanded by interleukin-2( IL-2) and Zol can get lots of purified γδT cells,and show cytotoxicity against the tumor cells of digestive system. γδT cells may be another important group of immune effector cells in the adoptive immunotherapy for cancer.

关 键 词：ΓΔT细胞 消化道肿瘤 免疫治疗 细胞毒活性 

分 类 号：R735.3[医药卫生—肿瘤]