人肺癌组织总RNA转染树突状细胞诱导特异性抗肿瘤免疫的研究  

Induction of specific cytotoxic T cells using dendritic cells transfected with human total RNA of lung cancer

作　　者：王坤[1] 吴一龙[1] 周清[1] 林嘉颖[1] 徐崇锐[1] 杨学宁[1] 黄少琼[1] 

机构地区：[1]广东省人民医院肿瘤中心广东省肺癌研究所,广东广州510080

出　　处：《中华肿瘤防治杂志》2008年第1期23-26,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：卫生部科学研究基金项目(98-2-377);广东省科技厅重点攻关项目(99M04903G;2KM04402S)

摘　　要：目的:探讨以人肺癌组织总RNA体外转染树突状细胞(dendriticcells,DCs)诱导细胞毒T淋巴细胞(cytotoxicTlymptocyte,CTL)发挥特异性抗肿瘤免疫的作用。方法:10例肿瘤组织经免疫组化测定为CEA、MUC1双阳性。一步法提取肿瘤组织总RNA和10例正常肺组织RNA。采集患者外周血单个核细胞体外诱导未成熟DCs,电穿孔法转染肿瘤和肺组织RNA,刺激DCs成熟后,部分行流式细胞仪检测,部分用于体外诱导CTL。以原代培养的肿瘤细胞和肺上皮细胞作为靶细胞,定量乳酸脱氢酶法测定杀伤率。结果:体外培养的DCs形态典型,高表达CD86、CD80、CD40、HLA-DR和CD83。转染肿瘤组织RNADCs的CEA和MUC1表达量为(20.53±3.64)%和(65.39±9.33)%,明显高于转染肺组织RNA的DCs[(0.78±0.39)%和(18.32±4.24)%],P<0.01。肿瘤组织RNA转染DCs诱导的CTL可产生针对肿瘤细胞的特异性杀伤;且用自身肿瘤组织RNA转染DCs诱导的CTL对自身肿瘤细胞杀伤活性最大。结论:以人肺癌总RNA体外转染DCs诱导CTL可以产生特异性抗肿瘤免疫,此方法构建肺癌疫苗是可行的。OBJECTIVE： To explore the specific antitumor immunity mediated by cytotoxic T lymphocytes （CTL） which were induced by dendritic cells （DCs） transfected with total RNA of lung cancer. METHODS： Tumor tissues and normal lung tissues of 10 patients with lung cancer were obtained from operation. CEA and MUC1 were positive in all tumor tissues. Total RNA were extracted from both tumor tissues and lung tissues. DCs were induced from peripheral blood monocytes in vitro and transfected with tumor total RNA or lung tissues RNA by electroporation. Some of these DCs were examined by flow cytometry analysis, others were used to induce CTL whose target cells were tumor cells and lung cells. RESULTS： DCs had a typical shape and expressed high level CD86, CD80, CD40, HLA-DR and CD83. CEA and MUC1 expressions on DCs transfected with tumor RNA were （20.53 ± 3.64）% and （65.39±9.33） %, which were much higher than those on DCs transfected with lung tissue RNA [（0.78 ± 0.39）% and （18.32±4.24）%], P〈0.01. CTL induced by DCs transfected with tumor RNA generated tumor-specific lysis and this effect was much more obvious when tumor RNA of oneself was used to transfect DCs. CONCLUSIONS： DCs transfected with total RNA of lung cancer can induce tumor-specific CTL response and specific antitumor immunity. Construction of DCs vaccine by this way is feasible.

关 键 词：肺肿瘤 树突状细胞 免疫治疗 

分 类 号：R734.2[医药卫生—肿瘤]