鸡传染性支气管炎病毒多表位核酸疫苗的构建及其免疫研究  

CONSTRUCTION AND IMMUNOGENICITY OF A MULTI-EPITOPE DNA VACCINE AGAINST INFECTIOUS BRONCHITIS VIRUS IN CHICKENS

作　　者：刘芳[1,2] 谭磊[2] 陆凤[2] 范瑾[2] 刘开春[2] 王欣[2] 廖瑛[2] 仇旭升[2] 孙英杰[2] 宋翠萍[2] 王桂军[1] 丁铲[2] 

机构地区：[1]安徽农业大学,合肥230036 [2]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国动物传染病学报》2015年第2期26-31,共6页Chinese Journal of Animal Infectious Diseases

基　　金：上海市科技兴农重点攻关项目(沪农科攻字(2013)第3-7号)

摘　　要：应用筛选出的鸡传染性支气管炎病毒(Infectious bronchitis virus,IBV)结构蛋白S1基因的T细胞抗原表位盒、Australia T株和M41株S1基因的B细胞抗原表位,定向克隆入真核表达载体p VAX1,构建成5个真核表达质粒:p VAX-S1T+M、p VAXS1B-M41、p VAX-S1B-T、p VAX-S1T+S1B-M41、p VAX-S1T+S1B-T。将每组提取制备的质粒溶液通过腿部肌肉多点注射免疫7日龄SPF雏鸡,28日龄时加强免疫1次,同时设立PBS组作为对照免疫组。分别在首免前及免疫后7、14、21 d以及二免后10 d翅静脉采血并分离血清,用间接ELISA方法检测抗体效价。二免后10 d用IBV病毒液攻毒测定保护率。ELISA抗体效价结果显示:二免后10 d各免疫组抗体水平达到最高,p VAX-S1B-T、p VAX-S1T+S1B-M41和p VAX-S1T+S1B-T组与PBS组相比差异均具有显著统计学意义(P<0.05),p VAX-S1、p VAX-S1T+M和p VAX-S1B-T组与PBS组相比差异具有极显著统计学意义(P<0.01)。攻毒结果表明,p VAX-S1、p VAX-S1B-T组的保护效率均为75%,高于PBS组(12.5%),表明表位抗原成分能够有效发挥保护效力,与S1全基因疫苗组保护率相当。本研究结果为研制安全有效的新型IBV疫苗提供了新的途径。The T cell epitope box of structural protein S 1 gene of avian Infectious bronchitis virus （IBV） and B cell epitopes of Australia T strain and M41 strain S1 gene were cloned into the eukaryotic expression vector pVAX1, constructing five groups of eukaryotic expression plasmids： pVAX- S 1T＋M, pVAX-S 1B-M41, pVAX-S 1 B-T, pVAX-S 1T＋S 1B-M41 and pVAX-S 1T＋S 1B-T. The plasmid solution from each group was administered to 7-days-old chickens via intramuscular route. Three weeks later, chickens were boosted with the same doses of DNA vaccines. Chickens receiving PBS served as controls. Blood samples were taken at 0, 7, 14 and 21 days post the first vaccination and 10 days post the second vaccination. Anti-IBV specific antibodies were measured in indirect ELISA. At 10 days post the second vaccination, chickens were challenged with virulent IBV strains. The result showed that antibody titers of chickens immunizedwith pVAX-S 1, pVAX-S 1T＋M and pVAX-S 1B-T were significantly higher （P〈0.01） than that of control group. The protection against the challenge was achieved in 75% chickens administered with pVAX-S I and pVAX-S 1B-T. In conclusion, the present study provided a new strategy to enhance the protection of IBV DNA vaccine.

关 键 词：传染性支气管炎病毒 抗原表位 核酸疫苗 免疫研究 

分 类 号：S852.659.6[农业科学—基础兽医学]