乙型脑炎病毒NS1蛋白亚单位疫苗制备及其免疫效果评价  

Manufacture of Japanese encephalitis virus NS1 protein subunitvaccine and evaluation of its immune effect

作　　者：钮俊俊 魏宁 朱硕 陈涛 杜冰洁 周登元 李秋燕 张路平 叶静[1] 司有辉 曹胜波[1] NIU Junjun;WEI Ning;ZHU Shuo;CHEN Tao;DU Bingjie;ZHOU Dengyuan;LI Qiuyan;ZHANG Luping;YE Jing;SI Youhui;CAO Shengbo(College of Veterinary Medicine,Huazhong Agricultural University,Wuhan 430070,China)

机构地区：[1]华中农业大学动物医学学院,湖北武汉430070

出　　处：《中国兽医学报》2023年第3期512-518,525,共8页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31825025,32022082,32030107,31972721,32002268)。

摘　　要：为制备乙型脑炎病毒(Japanese encephalitis virus,JEV)NS1蛋白亚单位疫苗并评价其免疫效果,将JEV NS1基因克隆至杆状病毒表达载体,构建重组杆状病毒。将重组杆状病毒感染High5细胞,收集培养基上清,并用镍柱亲和方法纯化NS1蛋白。将纯化的NS1蛋白与弗式佐剂制成亚单位疫苗,并通过小鼠免疫接种和攻毒模型评价该疫苗的免疫效果。结果表明,50,100,200μg/只剂量的亚单位疫苗免疫组的保护率分别为70%,80%,90%;同时,较非疫苗免疫组,NS1亚单位疫苗免疫可以显著降低小鼠脑组织中的病毒载量和炎症因子的表达,减轻小鼠的神经症状和脑部病理损伤。以上结果表明NS1蛋白作为亚单位疫苗具有较好的免疫保护效果,具有开发为JEV新型疫苗的潜力。In order to manufacture Japanese encephalitis virus(JEV)NS1 protein subunit vaccine and evaluate its immune effect,JEV NS1 gene was cloned into baculovirus expression vector to construct recombinant baculovirus.High5 cells were infected with recombinant baculovirus,and the NS1 protein was purified by nickel column.The subunit vaccine was produced by mixing the purified NS1 protein with Freund’s adjuvant,and the immune effect of the vaccine was evaluated by mouse model.The results showed that the protective rates of 50,100,200μg/mouse subunit vaccine against JEV challenge were 70%,80%and 90%,respectively.At the same time,compared with the un-immunized group,NS1 subunit vaccine immunization significantly reduced the viral load and the expression of inflammatory factors in the brain tissue of mice and attenuated the neurological symptoms and brain pathological damage of mice.Taken together,these results demonstrated that JEV NS1 subunit vaccine could stimulate effective immune response and could be used as a potential new vaccine for JEV.

关 键 词：乙型脑炎病毒 NS1蛋白 亚单位疫苗 杆状病毒表达系统 

分 类 号：S852.65[农业科学—基础兽医学] R535[农业科学—兽医学]