基于杆状病毒-昆虫细胞表达系统的猪脑心肌炎病毒样颗粒的表达及鉴定  

Expression and Identification of Porcine Encephalomyocarditis Virus-like Particles Based on Baculovirus-insect Cell Expression System

作　　者：王运航 景伟 穆素雨 孙世琪[1] 郭慧琛[1] 白满元 WANG Yunhang;JING Wei;MU Suyu;SUN Shiqi;GUO Huichen;BAI Manyuan(State Key Laboratory for Animal Disease Control and Prevention,College of Veterinary Medicine,Lanzhou University,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730000,China;School of Life Science,Yulin University,Yulin 719000,China)

机构地区：[1]中国农业科学院兰州兽医研究所兰州大学动物医学与生物安全学院动物疫病防控全国重点实验室,兰州730000 [2]榆林学院生命科学学院,榆林719000

出　　处：《畜牧兽医学报》2024年第10期4571-4578,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家重点研发计划(2022YFD1800603,2021YFD1800300);国家自然科学基金(32072859,32072847,32002272);科技人才与平台计划(202205AF150007);所长基金:《口蹄疫病毒持续性感染牛咽上皮细胞遗传变异的机制研究》LVRI-SZJJ-202107。

摘　　要：脑心肌炎(encephalomyocarditis)是由脑心肌炎病毒(encephalomyocarditis virus,EMCV)引起的以猪的脑炎、心肌炎为主要特征的病毒性人畜共患病,可感染多种哺乳动物,目前国内尚无有效疫苗。本研究旨在开发一款安全有效的疫苗以防控该病。采用P1、3CD共表达的构建策略,利用杆状病毒-昆虫细胞表达系统获得EMCV病毒样颗粒(virus-like particals,VLPs)并将其免疫小鼠,初步评价其在小鼠体内的免疫应答效果。结果显示,免疫组小鼠特异性抗体滴度达到1∶192以上,中和抗体滴度达到1∶64以上,且与ISA206佐剂混合使用可诱导机体产生更高的抗体水平。IL-4、IFN-γ、IL-5、TNF-α细胞因子水平均可达到20 pg·mL^(-1),且明显高于PBS组。综上,本研究获得的EMCV病毒样颗粒疫苗可以诱导机体产生良好的细胞免疫和体液免疫应答,还能诱导炎症细胞因子的产生,有助于激发机体的免疫反应,增强疫苗的保护效果,为EMCV的防控提供了理论依据和技术储备。Porcine encephalomyocarditis is a viral zoonotic disease caused by encephalomyocarditis virus and characterized by porcine encephalitis and myocarditis,which can infect a variety of mammals.Currently,there is no effective vaccine in China,so developing a safe and effective vaccine is an effective measure to prevent and control this disease.In this study,virus-like particals of porcine encephalomyocarditis virus were assembled by using the baculovirus-insect cell expression system with P1 and 3CD co-expression strategy,and the immune response effect of the virus in mice was preliminatively evaluated.The results showed that the specific antibody titer of mice in the immune group reached more than 1∶192,the neutralizing antibody titer reached more than 1∶64,and the combination with ISA206 adjuvant can induce higher antibody levels.The lev-els of IL-4,IFN-γ,IL-5 and TNF-αcytokines all reached 20 pg·mL^(-1),and were significantly higher than those in PBS group.Therefore,the EMCV virus-like particle vaccine obtained in this study can induce the body to produce good cellular and humoral immune responses,and can also induce the production of inflammatory cytokines,which helps to stimulate the body's immune re-sponse,enhance the protective effect of the vaccine,and provide theoretical basis and technical reserve for the prevention and control of EMCV.

关 键 词：杆状病毒-昆虫细胞表达系统 脑心肌炎病毒 病毒样颗粒 

分 类 号：S852.65[农业科学—基础兽医学]