目的探讨单纯疱疹病毒Ⅱ型(HSV-2)CTL表位疫苗对小鼠Th1/Th2型免疫应答的影响。方法用构建的HSV-2pVAX-gD-CTL重组质粒免疫BALB/c小鼠3次,末次免疫后第4周检测小鼠血清中gDIgG水平;MTT法检测小鼠脾淋巴细胞增殖情况;ELISA法检测脾淋巴细胞培养上清中INF-γ,IL-4水平;流式细胞仪测定小鼠CD4^+,CD8^+T淋巴细胞亚群。结果HSV-2pVAX-gD-CTL疫苗免疫小鼠后可以诱导脾淋巴细胞增殖及分泌INF-γ和IL-4,诱导CD4^+,CD8^+T细胞百分比的升高;并能刺激血清HSV-2gD特异性抗体的产生。其中pVAX-gD-CTL组在脾淋巴细胞刺激指数及其分泌INF-γ水平、CD4^+,CD8^+T细胞百分比升高方面均显著高于pVAX-gD对照组。结论CTL表位对单纯疱疹Ⅱ型重组DNA疫苗诱导的Th1型免疫应答有促进作用。
Objective To investigate the effect on Th1/Th2 response of HSV-2 g D DNA vaccine with CTL epitope. Methotis The recombinant plasmid pVAX-gD-CTL was used to inoculate BALB/C mice by intramuscular injection ( I. M. ) for three times. Four weeks after the last vaccinations, level of anti-HSV-2gD IgG antibody was detected. At the same time, spleen ceils of vaccinated mice were separated and stimulated with HSV-2gD protein. The proliferation of spleen lymphocytes was detected by MTT assay, the level of γ-IFN and IL-4 in cultural supernatant was detected by ELASA assay and the percentage of CD4^+, CD8^+T cell were detected by fluorescein activated cell sorter (FACS). Results After mice were inoculate by HSV-2 pVAX-gD-CTL vaccine, in both pVAX-gD-CTL and pVAX-gD groupe, the BALB/c mice could produce splenocytes proliferation and secreted IFN-γ and IL- 4,high percentage of CD4^+ , CD8^+ T lymphocyte and the antibody against HSV-2 gD. In pVAX-g D-CTL groupe, the level of splenocytes proliferation and secreted IFN-γ and IL-4, the percentage of CD4^+ , CD8^+ T lymphocyte were higher than that in the positive control group pVAX- gD, and the difference was significant. Conclusion CTL epitope recombinant DNA vaccine could specifically induced the Th1 immune response in BALB/C mice.