基质蛋白第221、226位氨基酸位点发生突变的重组水泡性口炎病毒构建和致病性研究  

Construction and pathogenesis of recombinant vesicular stomatitis virus with mutations at the amino acid sites of 221 and 226 in matrix protein

作　　者：吴昊 赵秋华 孙郭艳 柯勇 毕波 方心葵[1,2] 孙涛 WU Hao;ZHAO Qiuhua;SUN Guoyan;KE Yong;BI Bo;FANG Xinkui;SUN Tao(School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, China;Shanghai Municipal Veterinary Key Laboratory, Shanghai, 200240, China;Animal Disease Prevention and Control Center of Minhang District, Shanghai, 201109, China;Shanghai Pig Breeding Farm, Shanghai, 201408, China)

机构地区：[1]上海交通大学农业与生物学院,上海200240 [2]上海市兽医生物技术重点实验室,上海200240 [3]上海市闵行区动物疫病预防控制中心,上海201109 [4]上海种猪场,上海201408

出　　处：《畜牧与兽医》2018年第4期77-85,共9页Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金面上项目(31272562);上海市科委2015年度“科技创新行动计划”医学和农业领域科技支撑项目(15391901600)

摘　　要：重组水泡性口炎病毒（vesicular stomatitis virus,VSV）是一种具有良好应用前景的病毒载体,可用于制备疫苗和治疗肿瘤,但该载体可导致实验动物产生神经毒性,因此其使用仍存在安全性问题。为减低乃至去除野生型VSV的致病性,对该病毒基质蛋白M的第221、226位氨基酸进行了定点突变,制备了新型重组VSV,并围绕新型重组病毒的致病性展开了体外和体内试验。在体外试验中,221、226双位点突变重组VSV病毒（VSVM221,226）的复制能力相比野生型VSV（VSVXN2）降低可达20倍,且激发IFN-β达（871±1.3）pg/m L。在体内试验中,接种VSVM221,226小鼠体重下降仅约（8.9±0.2）%和（12.3±0.4）%,而接种VSVXN2小鼠体重下降高达（32.6±0.5）%和（30±2）%,且有死亡现象发生。试验结果表明,相比VSVXN2以及221、226单位点突变VSV,VSVM221,226的致病性显著降低,其致弱效应可能是由于病毒感染有效激活宿主细胞产生Ⅰ型干扰素所致。VSVM221,226有希望成为一个安全、有效的疫苗载体。Recombinant vesicular stomatitis virus（VSV） is an ideal viral vector for vaccine and viro-therapy; however,wild-type VSV still possesses toxicity in animals. When mice are injected with high-dosed VSV,they exhibit neurotoxicity. In order to reduce the pathogenicity of wild-type VSV,recombinant viruses aiming at amino acids of 221 and 226 in matrix protein were constructed with their pathogenicity characterized in vivo and in vitro. In the in vitro test,the ability of VSV with double mutations occurring at the amino acid sites of 221 and226（VSVM221,226） to replicate might be reduced by 20 folds,compared with the wild-type VSV（VSVXN2）,and the VSV might induce IFN-β by（871±1. 3） pg/m L. In the in vivo test,the body weight loss was only by（8. 9±0. 2） % and（12. 3±0. 4） % in the VSVM221,226 vaccinated mice,but（32. 6±0. 5） % and（30±2） % in the VSVXN2 vaccinated mice; some of the rodents even died. In contrast to VSVXN2,the pathogenicity of VSVM221,226 was significantly attenuated,which might be due to efficient induction of type I interferon in the host cells by viral infection. VSVM221,226 might be a promising vector for safe and effective vaccine development.

关 键 词：水泡性口炎病毒 基质蛋白 致病性 

分 类 号：S85[农业科学—兽医学]