乙型脑炎病毒DNA初免-蛋白加强策略在小鼠模型上的免疫效应评价  

Using mouse model to evaluate the immune effect of DNA prime-protein boost strategies targeting Japanese encephalitis virus

作　　者：于瑞明 田占成[1] 高闪电[1] 独军政[1] 关贵全[1] 殷宏[1,2] YU Ruiming;TIAN Zhancheng;GAO Shandian;DU Junzheng;GUAN Guiquan;YIN Hong(State Key Laboratory of Veterinary Etiological Biology,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,Gansu,China;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou 225009,Jiangsu,China)

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室,甘肃兰州730046 [2]江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《生物工程学报》2022年第8期2902-2911,共10页Chinese Journal of Biotechnology

基　　金：国家自然科学基金(31201899);中国农业科学院农业科技创新工程(CAAS-ASTIP-2016-LVRI)。

摘　　要：为了评价基因Ⅰ型乙型脑炎病毒prM-E DNA疫苗与prM和EⅢ融合抗原亚单位疫苗采用DNA初免-蛋白加强免疫策略对小鼠的免疫效果,本研究将prM-E融合基因插入到pVAX1真核表达载体中,构建重组表达载体prM-E-pVAX1作为DNA疫苗进行初免,利用原核表达系统获得的prM和EⅢ融合抗原作为亚单位疫苗进行加强免疫。将32只4-6周龄雌性BALB/c小鼠随机分成4组,设置prM-E-pVAX1 DNA疫苗组、DNA初免-蛋白加强免疫组、prM和EⅢ融合抗原亚单位疫苗组及pVAX1载体对照组,通过ELISA检测血清中特异性抗体水平;通过噬斑减少中和试验滴定中和抗体滴度;通过细胞因子表达丰度和淋巴细胞增殖试验分析不同疫苗免疫组诱导产生的细胞免疫反应。结果表明,用DNA初免-蛋白加强策略免疫的小鼠诱导产生的中和抗体滴度略高于prM和EⅢ融合抗原亚单位疫苗免疫组,显著高于prM-E-pVAX1 DNA疫苗免疫组。DNA初免-蛋白加强策略在小鼠模型中诱导产生了有效的Th1/Th2型免疫反应,特别是显著诱导了Th1型细胞免疫反应。本研究为预防流行性乙型脑炎提供了新的免疫策略和理论参考依据。In order to evaluate the immune effect of the genotypeⅠJapanese encephalitis virus prM-E DNA vaccine and the prM-EⅢfusion protein subunit vaccine on mice using DNA prime-protein boost strategy,the prM-E gene was inserted into the pVAX1 eukaryotic expression vector.The recombinant expression vector prM-E-pVAX1 was constructed as a DNA vaccine for initial immunity,and the recombinant prM-EⅢfusion protein was obtained using a prokaryotic expression system as a subunit vaccine for enhanced immunity.Thirty two female BALB/c mice aged 4-6 weeks were randomly divided into four groups,and a prM-E-pVAX1 DNA vaccine group,a DNA prime-protein boost immune group,a prM-EⅢsubunit vaccine group,and a pVAX1 vector control group were set up.The specific antibody level in serum was monitored by ELISA,the neutralizing antibody titer was detected by plaque reduction neutralization,and the cellular immune responses induced by different vaccine immune groups were analyzed by cytokine expression abundance and lymphocyte proliferation experiments.The results showed that the neutralizing antibody titers induced by mice immunized with the DNA prime-protein boost strategy were close to that of the group immunized with the single prM-EⅢsubunit vaccine,but significantly higher than that of the group immunized with the single prM-E-pVAX1 DNA vaccine.DNA prime-protein boost strategies induced effective Th1/Th2 immune responses in mouse models,in particular the Th1 cell-mediated immune responses.This study provides a new immune strategy that may facilitate the prevention of Japanese encephalitis.

关 键 词：流行性乙型脑炎 DNA疫苗 prM和EⅢ融合抗原 DNA初免-蛋白加强 

分 类 号：S852.4[农业科学—基础兽医学]