AN融合抗原制备及其加强BCG免疫效果的研究  

Construction of AN Fusion Antigen and Examination of Its Effect as BCG Immune Booster

作　　者：高笑笑 项智灏 陈福增[1] 张鹭[1] 刘军[1] GAO Xiaoxiao;XIANG Zhihao;CHEN Fuzeng;ZHANG Lu;LIU Jun(State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai 200438, China)

机构地区：[1]复旦大学生命科学学院,遗传工程国家重点实验室,上海200438

出　　处：《复旦学报（自然科学版）》2020年第6期677-684,共8页Journal of Fudan University：Natural Science

基　　金：国家科技重大专项(2018ZX10731301-001)。

摘　　要：亚单位疫苗研究中抗原的选择对疫苗效果至关重要.本研究选取结核分枝杆菌中ESX-Ⅰ~ESX-Ⅴ分泌系统的同源蛋白EsxA,EsxC,EsxE,EsxT,EsxN构建融合蛋白AN,通过序列分析、实验动物模型评估其作为亚单位疫苗的可行性.AN抗原的一级结构中存在332个T细胞受体强结合位点,说明融合抗原AN能够提供更多的T细胞识别表位.纯化后的AN抗原免疫C57BL/6小鼠,分离小鼠脾细胞进行胞内因子染色(ICS),结果证明AN抗原能激起高于或接近Ag85A的Th1型细胞因子分泌,具有较强的免疫原性,可以作为构建亚单位疫苗的候选抗原.利用BALB/c小鼠模型评估AN对于BCG免疫的加强保护效应,发现单次AN加强免疫,可使小鼠脾脏细菌清除率增加70.2%,增强了重组BCG的保护效果.The selection of antigens for construction of subunit vaccines is a critical factor for the success of such vaccines.In this study,we selected homologous proteins EsxA,EsxC,EsxE,EsxT,and EsxN in the ESX-Ⅰ—ESX-Ⅴsystem and constructed the fusion protein AN,and examined the potential of AN as a subunit vaccine in animal models.Sequence analysis revealed that AN contains 332T cell epitopes.Intracellular cytokine staining showed that C57/BL6 mice immunized with AN proteins secreted Th1 cytokines at levels higher or similar to Ag85a.In BALB/C mice,a single time boosting with AN decreased the bacterial burden by 70.2%.In summary,this study has constructed a fusion antigen AN and examined its potential as a subunit vaccine.

关 键 词：结核病 融合抗原 EsxA 

分 类 号：Q36[生物学—遗传学]