摘要
目的 :探讨转染hTCRVβ 8 4基因后健康人PBMC的免疫学特性及其对起源于乙肝病毒的肝癌细胞株BEL 74 0 2杀伤活性的改变。方法 :将hTCRVβ 8 4基因克隆至真核表达载体pcDNA3 1( )上 ,并转染健康人PBMC ,流式细胞术检测转染后PBMC中TCRVβ8 4蛋白表达 ,乳酸脱氢酶释放活性法检测重组质粒转染后PBMC对癌细胞BEL 74 0 2的杀伤活性。 结果 :TCRVβ8 4在基因转染后淋巴细胞中表达显著增高 ;与BEL 74 0 2共培养后 ,基因转染组CD3+ TCRVβ8 4T细胞增殖明显高于对照组 ;BEL 74 0 2G刺激后免疫细胞活化 ,表达CD12 2的细胞数量增多 ,表达CD19(B细胞活化的标志 )的B细胞增加 ;重组质粒转染后 ,PBMC对肝癌细胞BEL 74 0 2杀伤活性增强 ;透射电镜观察发现 ,重组质粒转染的PBMC使BEL 74 0 2凋亡。结论 :hTCRVβ 8 4基因修饰可显著增强淋巴细胞在超抗原BEL 74 0 2刺激下的增殖及免疫细胞活化 ,基因修饰后T淋巴细胞杀伤活性明显增强。
Objective:To investigate the immune state and the alteration of cytotoxicity to hepatocellular carcinoma cells BEL 7402 in hTCRV β 8 4 gene transfeced PBMC after co culturing with HBV derived hepatocellular carcinoma cells BEL 7402. Methods:The hTCRVβ 8 4 recombinant plasmid was transfered into peripheral blood mononuclear cells (PBMC).Flowcytometer analysis were used to assay the expression of TCRVβ8.4 gene and the the immune state of hTCRVβ8.4 gene modified PBMC.LDH release assay were used to test the cytotoxicity of the PBMC to hepatocellular carcinoma cells BEL 7402.Results:The expresstion of TCRVβ8.4 in CD3 +T cell was increased significantly after gene transfection.The percentage of TCRVβ8.4?CD122 + and CD19 +lymphocytes increased obviously after co cultured with BEL 7402,this indicates BEL 7402 stimulated the proliferation of these cells and cellular immunity.Transmission electronic microscope showed apoptosis in BEL 7402 induced by hTCRVβ8.4 gene transferred PBMC.LDH release assay reveals increased cytotoxicity of hTCRV β8.4 gene transfered PBMC to BEL 7402.Conclusion:The proliferation and the cytotoxicity were enhanced significantly in hTCRV β8.4 modified PBMC after stimulated by HBV derived hepatocellular carcinoma cells BEL 7402.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2003年第6期404-407,共4页
Chinese Journal of Immunology
关键词
TCRVβ8.4基因转染
PBMC
肝癌细胞
杀伤活性
hTCRV β8.4
Gene cloning and transfection
Lymphocytes
Hepatocellular carcinoma cells BEL 7402
