口蹄疫病毒非结构蛋白对豚鼠CD8^+T细胞应答的免疫增强作用  

Enhancement of CD8^+T cell immune response of nonstructural protein of foot-and-mouth disease virus in the immunized guinea pigs

作　　者：孙英军[1] 郑海学[1] 张艳[1] 靳野[1] 杨帆[1] 张志东[1] 刘湘涛[1] 

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学国家重点实验室农业部畜禽病毒学重点开放实验室国家口蹄疫参考实验室,甘肃兰州730046

出　　处：《中国兽医科学》2012年第10期1024-1030,共7页Chinese Veterinary Science

基　　金：甘肃省高层次人才科技创新创业扶持行动项目(1013JHTA008);公益行业专项(200903055-04;2008FY130100)

摘　　要：为了整体评价口蹄疫病毒非结构蛋白和前导蛋白对免疫应答的影响,选用反向遗传操作技术构建的与野生型毒株(Mya98/BY/2010)P1基因相同而非结构蛋白和前导蛋白不同的重组病毒(分别被命名为Re-Mya98/BY/2010)和Mya98/BY/2010)作为抗原,经BEI灭活后与4种佐剂乳化,制备不同的疫苗作为免疫原。然后免疫豚鼠,利用流式细胞仪检测免疫豚鼠CD4+T细胞和CD8+T细胞亚群的含量,ELISA方法检测免疫豚鼠血清中抗体应答。免疫后第28天进行攻毒试验。结果,抗原剂量相同的条件下,重组Re-Mya98/BY/2010疫苗比Mya98/BY/2010疫苗更能有效诱导CD8+亚群,每个时间点都含有较高的含量。ELISA结果显示,Mya98/BY/2010和Re-Mya98/BY/2010抗原都能诱导抗体应答,早期Re-Mya98/BY/2010诱导更高的抗体应答。豚鼠攻毒后,免疫Re-Mya98/BY/2010疫苗的保护率分别达到100%、83.33%、100%和50%,而免疫Mya98/BY/2010疫苗的保护率仅为20%、0、17%和25%,两者具有明显的统计学差异。结果表明,口蹄疫非结构蛋白在诱导免疫应答方面具有显著的诱导CD8+T淋巴细胞应答和提高免疫保护率的作用。To examine the role of nonstructural protein of foot-and-mouth disease virus（FMDV） in induction of protective immune response,the recombinant virus Re-Mya98/BY/2010 was constructed by replacing the structural protein-coding gene of the O vaccine strain,with that of the wild type strain Mya98/BY/2010.The immunogenicity of the recombinant and wild-type viruses was then investigated in immunized guinea pigs.Both the recombinant and wild-type viruses was inactivated with BEI and formulated with four different adjuvants（EMULSIGEN-BCL,EMULSIGEN-D,ISA206 and POLYGEN）.The guinea pigs were vaccinated muscularly and challenged with Mya98/BY/2010 on day 28 post-vaccination.Cellular and humoral immune responses on different days post-vaccination were then analyzed.In result,the stronger CD8＋ T cell and anti-FMDV antibody response was induced in guinea pigs vaccinated with the recombinant virus Re-Mya98/BY/2010 when compared to those in guinea pigs vaccinated the wild-type strain Mya98 / BY / 2010.The protection rates in guinea pigs vaccinated with the recombinant virus Re-Mya98 / BY / 2010with four difference adjuvants were 100%,100%,83.33%and 50%,respectively,while the guinea pigs vaccinated with FMDV Mya98 / BY / 2010were 20%,0%,17%and 25%,respectively.In conclusion,the recombinant FMDV Re-Mya98 / BY / 2010was able to induce both strong humoral and cellular immune responses,supporting the concept that a role of FMDV nonstructural protein in induction of the immune response.

关 键 词：口蹄疫病毒 非结构蛋白 细胞免疫 

分 类 号：S852.65[农业科学—基础兽医学]