细菌细胞表面展示体系提呈人源诺如病毒(GⅡ.4)抗原效果的评价  

Evaluation of Antigen Presentation of Human Norovirus(GⅡ.4) Using Bacterial Cell-Surface Display System in Mouse Model

作　　者：竺晖 王楠 鲁飞凤 张子蕾 唐峰 徐婷[1] 王大鹏[1] ZHU Hui;WANG Nan;LU Feifeng;ZHANG Zilei;TANG Feng;XU Ting;WANG Dapeng(Department of Food Science and Technology,School of Agriculture and Biology,Shanghai Jiao Tong University,Shanghai 200240,China)

机构地区：[1]上海交通大学农业与生物学院食品科学与工程系,上海200240

出　　处：《病毒学报》2020年第3期439-445,共7页Chinese Journal of Virology

基　　金：“十三五”国家重点研发计划(项目号:2017YFC1601200),题目:基于组学的食源性致病微生物快速高通量检测技术与装备研发。

摘　　要：人源诺如病毒(Human noroviruses,HuNoVs)是重要的肠胃炎病毒之一,造成了全球巨额经济负担,故其防控成为研究热点。探究不同形式的免疫原对机体体液和粘膜免疫的激发效果,成为HuNoVs疫苗研发的一个突破口。本研究利用前期以细菌细胞表面展示体系构建的假HuNoVs(GⅡ.4)和纯化的P(GⅡ.4)蛋白为对象,乳化后每隔两周分别进行Balb/C小鼠皮下多点和口服免疫,并以间接酶联免疫吸附试验测定血清中抗体的消长规律。同时,以磷酸盐缓冲液作为对照。抗体效价测定结果显示:皮下免疫组的假HuNoVs(GⅡ.4)可以有效激发小鼠体液免疫,其效价稍低于以P(GⅡ.4)蛋白作为抗原免疫获得的抗体。经过5次免疫后,血清中IgG抗体效价基本趋于稳定。但是,假HuNoVs(GⅡ.4)和P(GⅡ.4)蛋白的口服免疫组血清中均未测到IgA和IgG抗体。因此,细菌细胞表面展示体系假HuNoVs(GⅡ.4)可以作为HuNoVs疫苗候选抗原的提呈体系,为该病毒的疫苗研发提供借鉴。Human noroviruses(HuNoVs) are important foodborne viruses causing heavy economic burdens worldwide. Prevention and control of these viruses is a research hotspot. However,no vaccine was licensed so far. Finding a better way to present the major capsid antigens of HuNoVs is important. In this study,we evaluated the immunogenicity and efficacy of different antigens by inducing humoral immunity and mucosal immunity in a mouse model. The mimic HuNoVs,developed with bacterial cell-surface display system in our previous reports,and purified P(GⅡ.4)protein were used to immunize Balb/C mice via subcutaneous and oral routes. Mice were injected every 2 weeks with emulsion of antigen and adjuvant. The dynamics of antibody to P(GⅡ.4)in serum was detected by an indirect enzyme-linked immunosorbent assay. Phosphate-buffered saline was used as blank control. We demonstrated that humoral immunity was induced effectively by the mimic HuNoVs in the subcutaneous-multipoint group. The titer of immunoglobulin(Ig)G was lower than that induced by purified P(GⅡ.4). After five injections,the titer of antibodies became stable. However,IgA or IgG was not detected in the serum of mice in the oral-immunization group. Therefore,mimic HuNoVs could be a candidate vaccine to present the antigens of HuNoVs.

关 键 词：人源诺如病毒(HuNoVs) 细菌细胞表面展示体系 P蛋白 免疫 抗原提呈 

分 类 号：Q939.99[生物学—微生物学]