鼻咽癌患者外周血γδT细胞的体外扩增及对鼻咽癌细胞的体外杀伤活性

The ex vivo expansion of γ δ T cells from the peripheral blood of patients with nasopharyngeal carcinoma and their cytotoxicityto nasopharyngeals carcinoma lines in vitro

目的 :通过体外扩增鼻咽癌 (NPC)患者外周血γδT细胞 ,研究对NPC细胞体外细胞毒作用 ,以探讨γδT细胞在NPC免疫机制中的作用。方法 :用抗体固相法体外扩增 7例NPC患者 (NPC组 )及 6例健康者(对照组 )的外周血γδT细胞 ,以流式细胞仪检测γδT细胞亚型 ,用MTT法观察不同来源的γδT细胞在不同效靶比条件下 ,对Daudi、CNE1和CNE2细胞的细胞毒作用。结果 :对照组与NPC组外周血γδT细胞经体外扩增 ,纯度可达到 82 %以上 ,其中 90 %为Vγ9/Vδ2亚型 ;γδT细胞对Daudi、CNE 1和CNE 2细胞显示出作用强度相同的细胞毒作用 (P >0 .0 5 )。随效靶比升高 ,γδT细胞对CNE 1和CNE 2的细胞毒作用明显增强 ,在相同效靶比时对Daudi细胞的细胞毒作用最强。结论

Objective:γ δ T cells from the peripheral blood of nasopharyngeal carcinoma(NPC) patients were expanded in vitro and their cytotoxic activity to NPC lines was analysed. A role of γ δ T cell of immune mechanism in development of NPC was discussed.Method:The solid-phase monoclonal anti-TCR γ δ McAb was used to expand the γ δ T cells from peripheral blood of seven patients with NPC and six healthy donors. Then we detected the cytotoxicities of these γ δ T cell against tumour cell lines including Daudi Burkitt's lymphoma and the cell lines of NPC,CNE1 and CNE2 , at different effector :target (E∶T) ratio by MTT method.The immunophenotyping of γδ T cells was detected by flow cytometry.Result:γ δ T cell were obtained with a high purity (>82%) from healthy individuals and NPC patients in which 90% of expanded cells were Vγ9/Vδ2 subtype . The results of cytotoxicity assays revealed that activated γ δ T cells from both healthy donors and patients lysed Daudi,CNE1 and CNE2 cells at similar level(P> 0.05) when E∶T ratios increased,the level of cytotoxicity to each of the targets of CNE1 and CNE2 increased. These γ δ T cells exhibited highest level of cytotoxity to the Daudi cells in same ration of effector to target cell.Conclusion:γ δ T cell of NPC could significantly kill NPC in vitro.