用携有存活蛋白基因的重组腺病毒载体转染树突状细胞诱导细胞毒T细胞抗淋巴瘤免疫效应_医学论文.docVIP

用携有存活蛋白基因的重组腺病毒载体转染树突状细胞诱导细胞毒T细胞抗淋巴瘤免疫效应_医学论文 用携有存活蛋白基因的重组腺病毒载体转染树突状细胞诱导细胞毒T细胞抗淋巴瘤免疫效应_医学论文 作者：朱雄鹏，陈志哲，胡建达，李纯团，杨婷，许贞书【摘要】 　　本研究探讨转染survivin(SVV)基因的树突状细胞(DC)体外诱导特异性抗淋巴瘤的免疫效应。对人外周血DC进行诱导培养，以Ad转染DC，用Western blot 鉴定Survivin的表达，用乳酸脱氢酶(LDH)释放法检测细胞毒性T淋巴细胞（CTL）反应，用混合淋巴细胞反应（MLR）测定DC刺激同种异体淋巴细胞增殖的能力，以ELISA法检测上清中的细胞因子IL含量。结果表明：在MLR中，刺激应答比（S/R）为1∶5、1∶10、1∶50和1∶100时，转染Ad的DC有较强刺激同种异体淋巴细胞增殖的能力；转染Ad的DC组所诱发的CTL活性明显高于对照DC组；转染病毒后第2天，转染Ad的DC组的IL分泌水平高于对照DC组。结论：含存活蛋白基因的DC能够在体外诱导特异性CTL效应，对CA46细胞有明显的杀伤作用。 【关键词】 survivin基因 树突状细胞 CA46细胞系 淋巴瘤 细胞毒性T细胞 免疫治疗 　　Induction of Anti Abstract This study was purposed to investigate the immunological effects of modified dendritic cells (DCs) in inducing cytotoxic T cells (CTLs) effect against lymphoma cells. The DCs were derived from human peripheral blood and transfected with recombined adenovirus vector carrying survivin gene, Western blot was used to detect the expression of survivin, the lactate dehydrogenase (LDH) release test was used to determine the cytotoxicity of CTLs, the mixed lymphocyte reaction (MLR) was used to measure the ability to proleferate allolymphocyte by DCs, ELISA was used to assay IL12 level in supernatant. The results showed that the expression of survivin in transfected dentritic cells was confirmed by Western blot analysis. In MLR assay, DCs transfected with Adsurvivin could induce higher allogeneic lymphocytes reaction at the ratios of 1∶5,1∶10,1∶50 and 1∶100. DCs transfected with Adsurvivin had much higher activity of CTL to CA46 cells than control DCs. The levels of IL12 of supernatants containing DCs transfected with Adsurvivin were significantly higher than that in the control group. It is concluded that DCs transfected with Adsurvivin can induce CTL response in vitro against lymphoma cells. Key words survivin gene； dendritic cell； CA46 cell line lymphoma cytotoxic T cell immunotherapy 树突状细胞(dendritic cell, DC)作为机体内功能最强的专职抗原呈递细胞(APC)，在激活T细胞介导的免疫反应中起关键性作用，而survivin（存活蛋白，SVV）基因是肿瘤组织较为特异的基因，其编码的蛋白survivin成为肿瘤较理想的