负载hTERTC27基因的树突状细胞对胶质瘤细胞的杀伤效应

摘要

目的 探讨负载hTERTC27基因的树突状细胞(dendritic cells,DCs)治疗胶质瘤的可行性.方法 利用重组腺病毒(recombinant adenovirus,Adv)将hTERTC27基因转染到DCs,Western blot杂交鉴定hTERTC27蛋白的表达.实验分为3组:hTERTC27转染的DCs,单纯强化绿荧光蛋白(EGFP)转染的DCs和单纯的DCs.CCK8法检测各组DCs对淋巴细胞的增殖作用及诱导的细胞毒性T淋巴细胞(cytotoxic T lymphocytes,CTL)对胶质瘤细胞U87的杀伤活性,ELISA法检测T细胞上清中白介素-2(IL-2)和干扰素-γ(IFN-γ)的表达.结果 hTERTC27转染的DCs组对T细胞的增殖作用以及T细胞上清IL-2、IFN-γ的浓度均高于其余两组,当效靶比为40∶1时,这组T细胞对U87的杀伤率(50.4% ± 2.95%),也明显高于EGFP转染DCs组(38.0% ± 1.81%)以及单纯DCs组(33.7% ± 2.03%)(P ＝ 0.001).结论 hTERTC27基因转染的DCs能诱导CTL反应,对端粒酶阳性的U87具有显著的杀伤效应.%Objective To explore the feasibility of treatment with dendritic cells (DCs) transduced with hTERTC27 for glioma. Methods hTERTC27 gene was transduced to the DCs by recombinant adenovirus and hTERTC27 protein was detected using western blot. The DCs were divided into 3 groups : hTERTC27-transfected DCs. EGFP-transfected DCs and DCs only. T cells proliferation and cytotoxic T lymphocytes (CTL) response to U87 cells induced hy DCs were detected in each group using CCK8. The concentration of' interleukin-2 (IL-2) and interferon-γ(IFN-γ) in the supernatants of T cells were detected hy using ELISA. Results T cell proliferation and the concentration of IL-2 and IFN-γ were higher in the hTERTC27-transfected DCs than in the other groups. The cytotoxic rate against U87 in the hTERTC27-transfected DCs was (50.4％ ± 2.95％) at the ratio of effect / target cells of 40:1. which was much higher compared with EGFP-transfected DCs group (38.0％± 1.81％ ) and the DCs only group (33.7％ ± 2.03％) (ρ = 0.001). Conclusions hTERTC27 gene-transduced DCs can induce CTL response and has strong killing effect on hTERT+ U87 cells.