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目的 构建葡萄球菌肠毒素A(SEA)真核表达载体 ,并通过体内转染 ,观察其诱导抗小鼠肝癌的免疫效应。方法采用基因工程技术构建SEA基因的真核表达载体 ,使其表达SEA分子。通过阳离子脂质体介导的体内转染 ,观察其对小鼠肝癌的治疗作用。用51 Cr释放法测定治疗组与对照组动物脾淋巴细胞杀伤H2 2 细胞的活性。结果成功地构建了SEA真核表达载体 pLXSN SEA ,体内转染 pLXSN SEA的荷瘤小鼠肿瘤明显缩小 ,生存期延长 ,4 10小鼠肿瘤完全消退 ,且长期无瘤生存。CTL杀伤活性实验表明 ,瘤区内转染pLXSN SEA可诱导强烈的CTL杀伤效应 ,与对照组相比较差异显著 (P <0 .0 1)。结论超抗原SEA体内转染对肿瘤具有明确的治疗作用 ,有望用于抗肿瘤的生物治疗
Objective To construct the eukaryotic expression vector of staphylococcal enterotoxin A (SEA), and to observe its immune effect induced by anti-mouse hepatoma by transfection in vivo. Methods The eukaryotic expression vector of SEA gene was constructed by genetic engineering to express SEA molecule. Cationic liposome-mediated transfection in vivo was used to observe the therapeutic effect on mouse liver cancer. The activity of splenic lymphocytes killing H2 2 cells in treated and control animals was determined by 51 Cr release assay. Results The SEA eukaryotic expression vector pLXSN SEA was successfully constructed. Tumor-bearing mice transfected with pLXSN SEA in vivo were significantly reduced in tumor size and prolonged in survival time. Tumors in 41 of mice completely regressed and had long-term tumor-free survival. CTL cytotoxicity assay showed that transfection of pLXSN SEA in the tumor area could induce a strong CTL killing effect, which was significantly different from the control group (P <0.01). CONCLUSIONS: Transfection of superantigen SEA in vivo has a definite therapeutic effect on tumors and is expected to be used in anti-tumor biotherapy