基于酿酒酵母表面展示技术构建寨卡口服疫苗及其免疫活性初探

Construction of ZIKV Oral Vaccine Based on Saccharomyces cerevisiae Surface Display Technology and the Preliminary Study on Its Immune Activity

目的:基于酿酒酵母表面展示技术构建具有免疫活性的寨卡病毒(ZIKV)口服疫苗。方法:以Zika/SZ01/2016的包膜(Envelope)基因作为研究对象,以酿酒酵母表面展示质粒p YD1为骨架,构建EBY100/p YD1-Envelope。通过蛋白质印迹法(western blot)、免疫荧光分析和流式细胞仪分析,对Envelope蛋白进行定性分析,通过BCA蛋白测定试剂盒对Envelope蛋白进行定量分析。以SPF级的BALB/c小鼠作为动物模型,考察3种免疫方案(给药方案A:50 OD_(600 nm)·d^(-1)×3 d,给药方案B:75 OD600 nm·d^(-1)×2 d,给药方案C:150 OD_(600 nm)·d^(-1)×1 d)的口服免疫效果。结果:3种免疫方案均能诱发产生较高水平的体液免疫应答和黏膜免疫应答。给药方案C的免疫效果最佳。结论:本研究首次探讨了ZIKV口服疫苗的免疫效果,为有效预防ZIKV感染提供了一种可供参考的解决方案。

Objective： To construct Zika virus（ ZIKV） oral vaccine based on the Saccharomyces cerevisiae（ S. cerevisiae） surface display technology. Methods： EBY100/pYD1-Envelope was constructed based on the Envelope gene of Zika/SZ01/2016 used as research subject and S. cerevisiae surface display plasmid pYD1 was used for a backbone. Qualitative analysis was performed by western blot,immunofluorescence assay and flow cytometric assay; quantitative analysis was then made by BCA protein detecting kit as well. SPF grade BALB/c mice were used as animal model to study the oral immune effect of 3 dosage regimens（ Regimen A： 50 OD（600 nm）·d（-1）× 3 d,Regimen B： 75 OD（600 nm）·d（-1）× 2 d,Regimen C： 150 OD600 nm·d（-1）× 1 d）. Results： All the three regimens could induce higher levels of humoral immune response and mucosal response. Regimen C was preferred. Conclusion： In this study,it is the first time to evaluate the immune efficacy of ZIKV oral vaccine. Moreover,it provides a reference strategy for effective prevention of ZIKV infection.