Rv3129的构建、表达、纯化以及免疫原性分析  

Construction,Expression,Purification and Immunogenicity Analysis of Rv3129

作　　者：王晓春[1] 王艳 许礼发[1] 毛莉蓉 孔令雲 张子安 WANG Xiaochun;WANG Yan;XU Lifa;MAO Lirong;KONG Lingyun;ZHANG Zian(School of Medicine,Anhui University of Technology,Huainan Anhui 232001,China)

机构地区：[1]安徽理工大学医学院,安徽淮南232001

出　　处：《安徽理工大学学报（自然科学版）》2022年第6期87-93,共7页Journal of Anhui University of Science and Technology:Natural Science

基　　金：安徽省高校自然科学研究重点资助项目(KJ2015A093,KJ2016A211)。

摘　　要：目的以结核分枝杆菌休眠期相关蛋白Rv3129作为研究对象,构建其重组质粒并进行原核表达,并通过体外人群实验、体内动物实验验证其免疫原性。方法构建、表达并验证Rv3129蛋白,通过人外周血刺激释放实验、小鼠血清抗体水平检测以及各细胞因子检测实验验证该疫苗的免疫原性。结果成功构建、表达及纯化rRv3129,可刺激M.tb感染人群外周血能够有效分泌高水平IFN-γ。动物实验中,初免增强策略下的亚单位疫苗Rv3129/DIM能够诱导强烈的Th1型免疫应答,分泌的抗体水平显著高于其他组别。结论该亚单位疫苗具有较强的免疫原性,能够在机体内诱导强烈的Th1型免疫反应,对预防成人结核病可能具有重要意义。Objective To construct a recombinant plasmid and perform prokaryotic expression of Rv3129,a dormancy-associated protein of Mycobacterium tuberculosis,and to verify its immunogenicity by in vitro population experiments and in vivo animal experiments.Methods The Rv3129 protein was constructed,expressed and validated in vivo,and the immunogenicity of the vaccine was verified by human peripheral blood stimulated release assay,mouse serum antibody level assay and various cytokine assay assays.Results Successful construction,expression and purification of rRv3129 stimulated the peripheral blood of M.tb-infected population to secrete high levels of IFN-γ.In animal experiments,the subunit vaccine Rv3129/DIM under the primary immune booster strategy induced a strong Th1-type immune response and secreted significantly higher levels of antibodies than other groups.Conclusion The subunit vaccine has strong immunogenicity and can induce strong Th1-type immune responses in the organism,which is important for anti-tuberculosis and protection in adults.

关 键 词：结核分枝杆菌 亚单位疫苗 卡介苗 复合佐剂 初免增强策略 

分 类 号：R378.911[医药卫生—病原生物学]