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目的 :研究转 4 1BBL基因小鼠肝癌细胞疫苗体外诱导淋巴细胞特异性杀伤活性及刺激同系小鼠脾细胞产生细胞因子 (IL 2、TNF α和GM CSF)的能力 .方法 :采用脂质体介导法将真核表达质粒pCDNA3.1(+) m4 1BBL导入小鼠肝癌细胞Hepa1 6 ,经G4 18筛选后获得稳定高表达克隆 ,以丝裂霉素C(MMC)处理后 ,制成肿瘤细胞疫苗 (TCV) ,经体外与同系小鼠脾淋巴细胞共同培养后 ,测定淋巴细胞特异性杀伤活性及对脾细胞产生细胞因子 (IL 2、TNF α和GM CSF)的影响 .结果 :转染 4 1BBL的Hepa1 6细胞能够高表达 4 1BBL蛋白 ,并且经MMC处理后制成的瘤苗在培养 4 8h仍能表达m4 1BBL .与野生型的Hepa1 6细胞相比 ,上述瘤苗能诱导淋巴细胞产生针对亲本的小鼠肝癌细胞Hepa1 6的特异性杀伤作用 (P <0 .0 5 ) ,但是对于小鼠肝癌细胞H2 2及成纤维细胞NIH3T3无效 ;此瘤苗在体外能显著增强脾细胞分泌细胞因子IL 2、TNF α和GM CSF的能力 .结论 :转 4 1BBL基因小鼠肝癌细胞疫苗能诱导有效的抗肝癌免疫反应 .
Objective: To investigate the ability of transfection of 4 1BBL mouse hepatoma cell vaccine induced lymphocyte-specific cytotoxicity (IL 2, TNF α and GM CSF) in spleen cells of syngeneic mice in vitro.Methods: Liposome mediated The recombinant plasmid pCDNA3.1 (+) m4 1BBL was transfected into Hepa1 6 hepatoma cell line HepG6, which was screened by G418 to obtain stable and highly expressed clones. After treated with mitomycin C (MMC), the cells were made into tumor cells (TCV) were cultured in vitro and splenic lymphocytes of syngeneic mice to determine the specific cytotoxicity (IL 2, TNF α and GM CSF) of lymphocyte.Results: 1BBL Hepa1 6 cells were highly expressed 4 1BBL protein, and the MMCs treated with MMC could still express m4 1BBL after 48 h culture.Compared with wild-type Hepa1 6 cells, the above-mentioned vaccine can induce lymphocyte production Specific killing effect on Hepa1 6 mouse hepatoma cells (P <0 05), but not on H 2 2 and NIH 3T3 fibroblasts in mice; this vaccine can significantly enhance the secretion of spleen cells in vitro Factor IL 2, TNF α GM CSF ability Conclusion: Tumor Vaccine 4 1BBL turn transgenic mice can induce an effective immune response against liver cancer.