超抗原葡萄球菌肠毒素A对肝癌细胞免疫原性影响的研究  

Influence of staphylococcal enterotoxin A on immunogenicity of hepatocellular carcinoma cells

作　　者：李增山[1] 隋延仿[1] 姜永强[2] 雷祚荣[2] 

机构地区：[1]第四军医大学病理教研室肿瘤抗原肽实验室,西安710032 [2]军事医学科学院微生物流行病研究所

出　　处：《中华肝脏病杂志》2001年第6期343-344,共2页Chinese Journal of Hepatology

基　　金：国家自然科学基金(39770827)

摘　　要：目的给肝癌细胞转入超抗原分子葡萄球菌肠毒素A(staphylococcal enterotoxin A,SEA)的编码基因,使其表达这种毒素分子,以增强肝癌细胞的免疫原性,达到增强肝癌细胞诱导免疫排斥反应的目的.方法构建SEA编码基因的逆转录真核表达载体,转染病毒包装细胞系PA317后,行滴度测定.以高滴度克隆培养上清转导肝癌细胞株HHCC,筛选获得阳性表达株.然后利用外周血混合淋巴细胞进行杀伤实验.结果成功的构建了SEA的逆转录真核表达载体pLXSN-SEA,转导肝癌细胞后,获得表达SEA分子的阳性克隆HHCC-SEA.培养上清中SEA含量在pg的水平.外周血混合淋巴细胞杀伤实验结果显示,T淋巴细胞对转导后肝癌细胞的杀伤率可达45.6%.高于对HHCC 20.7%的杀伤率,T细胞对转导后肝癌细胞杀伤反应的Km值为5.18×104,而HHCC组的Km值为2.92 × 105,与前者的差别有统计学意义.结论肝癌细胞在转入SEA分子的编码基因后,表达出微量的SEA蛋白,却具有较强的免疫激活能力.即表达SEA的肝癌细胞在很低的浓度下,激活的T细胞即可对其达到最大杀伤反应速度,对T细胞的激活能力远远高于HHCC.Objective To transduce the gene of staphylococcal enterotoxin A（SEA) into a HCC cell line HHCC and to make the cells to express the molecules of SEA for the purpose of improving the immunogenicity and inducing im-mune rejection of HCC. Methods Construct retroviral vector containing the gene of SEA and tranduce it into HHCC cells. Then the integration, transcription and expression of recombinant were analyzed. At last cytotoxicity test of peripheral blood mixed lymphocytes was carried out. Results We obtained the retroviral vector pLXSN-SEA, transduced into HHCC cells and obtained the HCC cell line that expressed SEA toxin protein. The content of SEA in supernatant was at the level of pg. The cytotoxicity of HHCC transduced with SEA by T lymphocytes was 45.6%, which was higher than that of HHCC(20.7%). The Km value of T lymphocytes against HHCC transduced with SEA and without SEA was 5.18 ×10~4 and 2.92 ×10~5, separately. The difference had statistical significance. Conclusions After transducing the SEA gene, although tiny quantity of expres-sion is detected, the cytotoxicity assay proves that the expression of SEA lead to a robust immune response.

关 键 词：肝癌 超抗原分子葡萄球菌肠毒素A 肝癌细胞 免疫 

分 类 号：R735.7[医药卫生—肿瘤]