基因转染Sp2/0细胞作为检测乙肝病毒基因疫苗细胞免疫效应的靶细胞的研究

Transfected sp2/0 cells as target cells to investigate cellular immune responses in mice induced by hbv gene vaccine

目的研究乙型肝炎病毒(HBV)S基因转染的Sp2/0细胞作为检测HBV基因疫苗细胞免疫效应的靶细胞的可行性。方法以脂质体介导基因转染,含G418的培养基筛选,获得HBV基因S转染的Sp2/0细胞;构建编码HBV抗原蛋白S的重组真核表达质粒pCR3.1-S作为HBV基因疫苗;采用51Cr 4 h 释放法体外检测HBV基因疫苗免疫接种小鼠的淋巴细胞杀伤功能。结果与空载体对照组相比较,HBV基因疫苗诱发Balb/c 小鼠产生较好的HBV特异的细胞免疫应答(P<0.05)。结论以Sp2/0基因转染细胞作为靶细胞检测免疫BALB/c小鼠的淋巴细胞杀伤功能是可行的。

Objective: To study the possibility of gene-transfected Sp2/0 cells as target cells to measure cytotoxic T lymphocyte activity induced by HBV S gene vaccine. Methods: Sp2/0 cells were transfected with recombinant plasmid of pCR3.1-S as target cells. Recombinant eukaryotic expression plasmid of pCR3.1-S encoding S protein of hepatitis B virus was constructed as HBV gene vaccine. Then cellular immune response to HBV could be detected by 4-hour 51Cr release assays in BALB/c mice inoculated with HBV gene vaccines. Results: Compared with plasmid vectors, HBV gene vaccine could induce strong cytotoxic T lymphocyte activities in immunized mice (P<0.05). Conclusions: It is possible that Sp2/0 cells are transfected with genes of HBV as target cells to measure cytotoxic T lymphocyte activity in immunized BALB/c mice.