目的：为探索逆转录病毒载体在免疫细胞中的表达和基因治疗中的应用。方法：用DNA重组技术构建乙肝病毒表面抗原C(HBV-S)基因重组逆转录病毒载体，电穿孔转染PA317后，筛选高表达克隆，用假病毒颗粒感染HepG2、巨噬细胞(RAW264.7)和EL4细胞，分别用RT-PCR及ELISA法检测目的基因表达。结果：HBsAg在上述细胞中获得不同程度的表达，48h细胞上清中HBsAg含量（A值）分别为0.92、0.53、0.42。结论：本实验所用载体，其目的基因可在细胞内稳定表达，而且，在巨噬细胞等抗原提呈细胞中均有较高的表达，这提示我们质粒免疫有可能通过刺激巨噬细胞诱导机体产生较强的体液免疫和细胞免疫。作为一种高效的基因转移系统，该表达系统可用于基因免疫和基因治疗。%AIM： To investigate the effectiveness of recombinanted retrovirusvector in gene therapy. METHODS: The retroviral vector pLXSN-S was constructed and transferred into PA317 by means of electroporation, then HepG2、RAW264.7 and EL4 cells were infected with the pseudovirus produced from PA317, which highly expressed HBsAg. HBsAg expression was tested by RT-PCR and ELISA. RESULTS: HBsAg was expressed variously in the eukaryotic cells mentioned above. HBsAg ( A value) of the cell supernatants (48 hours) were 0.92，0.53，0.42, respectively. CONCLUSION: The vector used in this study was an effective vector to carry genes of interest to target cells and macrophage, and high level HBsAg was expressed in antigen presenting cell such as macrophage, It indicated that plasmid immunity can induce the B lymphocyte and T lymphocyte response to hepatitis B virus surface antigen by stimulating macrophage. As a vector, it may be useful in the test for gene immunity and gene therapy.
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