表达非洲猪瘟病毒p54蛋白的重组猪瘟兔化弱毒疫苗的构建及其免疫原性评价  

Generation and immunogenicity evaluation of a recombinant classical swine fever virus C-strain expressing p54 protein of African swine fever virus

作　　者：李树稳 杨晓柯 管翔雨 张广柱 黄淑坚 李永锋[2] 仇华吉 LI Shuwen;YANG Xiaoke;GUAN Xiangyu;ZHANG Guangzhu;HUANG Shujian;LI Yongfeng;QIU Huaji(College of Life Science and Engineering,Foshan University,Foshan,Guangdong 528231,China;State Key Laboratory of Animal Disease Prevention and Control,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150069,China;Harbin Weike Biotechnology Co.,Ltd.,Harbin 150069,China)

机构地区：[1]佛山科学技术学院生命科学与工程学院,广东佛山528231 [2]中国农业科学院哈尔滨兽医研究所动物疫病防控国家重点实验室,黑龙江哈尔滨150069 [3]哈尔滨维科生物技术有限公司,黑龙江哈尔滨150069

出　　处：《中国兽医学报》2024年第10期2094-2100,2122,共8页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31972673)。

摘　　要：通过同源重组构建pHCLV-p54感染性克隆,将其转染至SK6细胞盲传拯救出重组病毒rHCLV-p54,并测定其在体外的遗传稳定性、生长曲线,同时免疫家兔评价其免疫效果。结果显示,E183L基因能够在重组病毒中稳定遗传,但是插入的外源基因影响C株的复制;家兔免疫试验结果显示,重组病毒rHCLV-p54能够诱导家兔产生抗ASFV的特异性抗体。结果表明,表达非洲猪瘟病毒(African swine fever virus, ASFV)p54蛋白的重组病毒rHCLV-p54具有良好的免疫原性,有望作为候选疫苗进一步评价。African swine fever(ASF)is a virulent,hemorrhagic disease of swine caused by African swine fever virus(ASFV),which seriously affects the healthy development of Chinese pig industry.The genome of ASFV is large and encodes more than 165 proteins.Among them,the p54 protein is encoded by the E183L gene,which has various functions such as participating in viral assembly,inducing apoptosis and inducing immune response.The conventional Chinese vaccine(C-strain)is a safe and effective attenuated vaccine developed by Chinese scientists.It can efficiently protect against attacks from various genotypes of classical swine fever virus(CSFV).The aim of this study was to investigate whether C-strain can express ASFV p54 protein serve as a delivery vector for ASF genetically engineered vaccines.An infectious clone of pHCLV-p54 was constructed by homologous recombination,the recombinant virus rHCLV-p54 was rescued by transfecting it into SK6 cells by blind passaging.Its genetic stability and growth curve were determined in vitro,while rabbits were immunized to evaluate its immunity effect.The results showed that the E183L gene remained genetically stable in the recombinant virus,indicating that the E183L gene could be stably inherited in recombinant viruses,but the inserted exogenous gene affected the replication of the C-strain.The results of the rabbit immunization test showed that the recombinant virus rHCLV-p54was able to induce ASFV-specific antibodies.The above results indicated that we have successfully constructed a recombinant C-strain that stably expresses the ASFV p54protein.In summary,the recombinant virus rHCLV-p54has a good immunogenicity and is warranted for further evaluation as a vaccine candidate.

关 键 词：非洲猪瘟 猪瘟 猪瘟兔化弱毒疫苗 活载体疫苗 

分 类 号：S852.65[农业科学—基础兽医学]