CAJ | 학술논문

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。
목적：위탐색역전록병독재체재면역세포중적표체화기인치료중적응용。방법：용DNA중조기술구건을간병독표면항원C(HBV-S)기인중조역전록병독재체，전천공전염PA317후，사선고표체극륭，용가병독과립감염HepG2、거서세포(RAW264.7)화EL4세포，분별용RT-PCR급ELISA법검측목적기인표체。결과：HBsAg재상술세포중획득불동정도적표체，48h세포상청중HBsAg함량（A치）분별위0.92、0.53、0.42。결론：본실험소용재체，기목적기인가재세포내은정표체，이차，재거서세포등항원제정세포중균유교고적표체，저제시아문질립면역유가능통과자격거서세포유도궤체산생교강적체액면역화세포면역。작위일충고효적기인전이계통，해표체계통가용우기인면역화기인치료。
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.