AFP基因转染CD34^+造血干细胞来源的树突状细胞诱导特异性抗肝癌免疫  

Induction of specific immune response against hepatocellular carcinoma by AFP gene transfected dendritic cells from CD34 + hematopoietic stem cells

作　　者：贾军[1] 任军[1] 张利旺[1] 张红梅[1] 郭晓彤[1] 斯晓明[1] 王九花[1] 程婕[1] 

机构地区：[1]第四军医大学西京医院肿瘤科,陕西西安710033

出　　处：《第四军医大学学报》2005年第8期755-757,共3页Journal of the Fourth Military Medical University

基　　金：教育部全国高等院校优秀青年教师奖励计划资助项目(TRAPOYT99 016)

摘　　要：目的: 以AFP基因转染CD34+造血干细胞来源的树突状细胞(DC),诱导DC产生特异性抗肝癌免疫,观察其对肝癌细胞的杀伤.方法: 采用化疗和集落刺激因子动员肝癌患者外周血造血干细胞,血细胞分离机采集CD34+造血干细胞,IL 4、GM CSF联合TNF α刺激CD34+细胞分化为DC.通过脂质体转染的方法将携带人AFP基因质粒pEGFP AFP转染树突状细胞,MTT法检测对人肝癌细胞HepG2的特异性杀伤.结果: 经细胞因子刺激诱导培养的DC具有典型的分枝状突起,高表达CD11C,CD80,CD86.转染AFP基因后可见到转染细胞表达绿色荧光蛋白,细胞化学染色胞质内可见到红色颗粒.MTT法检测AFP基因转染后的DC可诱发特异性CTL反应杀伤HepG2细胞,AFP基因转染组、空载体组和对照组的杀伤率分别为( 92 0±3 5 )%, ( 75 5±4 1 )%和(72 9±3 4)%,转染组与空载体组和对照组间存在显著性差异(P<0 05).结论: AFP基因转染CD34+造血干细胞来源的DC可诱导特异性CTL反应杀伤肝癌细胞.AIM: To transfect dendritic cells originating from CD34 + hematopoietic stem cells with AFP gene and to observe the immune response against hepatocellular carcinoma.METHODS: CD34 +hematopoietic stem cells were harvested after mobilization by chemotherapy and G-CSF.CD34 + hematopoietic stem cells were induced to differentiate into dendritic cells by cytokine IL-4,GM-CSF and TNF-α.At the fifth day of culture,plasmid pEGFP-AFP was transfected into dendritic cells using lipofectamine 2000.The immune response triggered by dendritic cells was valued by MTT method.RESULTS: The dendritic cells originated from CD34 + cells had typical dendritic stick and expressed CD11C,CD80 and CD86.After transfected with AFP gene,the dendritic cells expressed green fluorescence protein and cytochemical assay showed that there were red spots in the cytoplasm of dendritic cells.MTT method detected the cytotoxic T-lymphocyte (CTL)reaction against HepG2 cells and showed that the transfected dendritic cells could induce AFP specific immune response.The killing rates of CTL response were respectively (92.0 ±3.5)%,(75.5 ±4.1)% and (72.9 ±3.4)% in AFP gene transfected group,pEGFP transfected group and control group.There was a significant difference between the AFP gene transfected group and the other two groups.CONCLUSION: AFP gene transfected dendritic cells originating from hematopoietic stem cells can induce specific immune response and stimulate antigen specific CTL response to kill AFP-expressed hepatocellular carcinoma cells.

关 键 词：甲胎蛋白类 转染 树突细胞 肝肿瘤 

分 类 号：R739.41[医药卫生—肿瘤]