单纯疱疹病毒II型CTL表位DNA疫苗的Th1/Th2免疫应答研究

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。

Study on Th1/Th2 Response of HSV-II CTL Epitope DNA Vaccine

Objective To investigate the effect on Th1/Th2 response of HSV-2 g D DNA vaccine with CTL epitope.Methods 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 cells 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.