重组PA4复制子病毒颗粒疫苗免疫原性的分析

摘要

Objective: To construct recombinant Semliki forest virus(SFV) replicon virus particles contained domain 4 of the protective antigen (PA4) from anthrax,and further study its immunogenicity. Methods: The recombinant virus particles (RVP) were prepared by cotranfecting the expression vector pSCAR-SPA4 with helper vector pSHCAR and expression of antigen PA4 was also confirmed by indirect imumofluorescence. Mice were immunized with RVP and antibody level of sera from mice was measured. Lymphocyte proliferate responses and cytokine levelswere measured after specifically stimulate splenocyte. Results: After immunization of mice,high antibody titer was measured. Specifically stimulated splenocytes with PA4 antigen can induce lymphocyte proliferation and secretion of IFN-γ and IL-4. Conclusion: Recombinant PA4 replicon virus particles vaccine can induce high titer antibody and cellular immunity response. The data presented in the report suggested that the recombinant PA4 replicon virus particles in this study may be developed into an alternative vaccine candidate for anthrax.%目的:构建携带炭疽毒素保护性抗原第四结构域(PA4)基因的重组Semliki森林病毒(SFV)复制子病毒颗粒,并对其免疫原性进行研究.方法:将编码炭疽PA4的SFV复制子DNA载体pSCAR-SPA4,与辅助SFV DNA载体pSHCAR共转染BHK21细胞,制备表达PA4的重组复制子病毒颗粒；用重组复制子病毒颗粒疫苗免疫小鼠,并采用ELISA法检测其血清抗体水平和细胞因子IFN-γ和IL-4.结果:免疫小鼠血清中检测到较高的抗体水平,免疫小鼠的脾淋巴细胞经特异性抗原刺激后产生了明显的T细胞增殖反应并分泌产生了IFN-γ和IL-4.结论:重组PA4复制子病毒颗粒疫苗免疫小鼠后能够产生特异性的抗体反应和细胞免疫反应.制备的重组PA4复制子病毒颗粒极有潜力作为人用炭疽候选疫苗,为进一步研究新型炭疽疫苗奠定了基础.