禽流感病毒H5亚型主要保护性抗原在禽痘病毒中的高效表达  

Recombinant fowlpox virus expressing HA from subtype H5N1 of avain influenza virus and its protective efficacy

作　　者：陈素娟[1] 孙蕾[1] 石火英[1] 彭大新[1] 刘秀梵[1] 

机构地区：[1]扬州大学农业部畜禽传染病重点开放实验室,江苏扬州225009

出　　处：《中国兽医学报》2007年第2期200-202,共3页Chinese Journal of Veterinary Science

基　　金：国家科技攻关资助项目(2004BA519A02);国家"十五"科技攻关资助项目(2004BA519A19))

摘　　要：将H5亚型禽流感病毒血凝素HA基因克隆入插入载体p11S中获得重组转移质粒p11SH5A,通过酶切鉴定获得了预期的转移质粒p11SH5A,将质粒p11SH5A和野生禽痘病毒(wtFPV)共转染鸡胚成纤维细胞(CEF),通过蓝白斑筛选纯化得到重组病毒rFPV-11SH5A。以间接免疫荧光法证实,HA基因得到了表达。将该重组病毒rFPV-11SH5A以105PFU/只免疫7日龄SPF鸡,于7、10、14、18、21d分别采血分离血清检测HI抗体,于免疫21d后用105ELD50的野生病毒进行肌肉注射观察疫苗保护率。结果表明,该疫苗能提供100%的保护。The hemagglutinin(HA) gene from subtype H5N1 avain influenza virus was directionally inserted into transferring vector p11S,resulting in recombinant transferring vector p11SH5A.Then the recombinant transferring vector p11SH5A was used to transfect the chicken embryo fibroblasts(CEF).Which was pre-infected with wild type fowlpox virus,to generate recombinant fowlpox virus expressing H5N1(rFPV-11SH5A).By selection of blue plaques on the CEF,over laid with agar containing X-gal,rFPV-11SH5A was obtained and purified.The in vitro expression of HA by rFPV-11SH5A was detected in the recombinant virus-infected CEF with indirect immunofluorescence.Experiments on chickens demonstrated that rFPV-11SH5A had the same protective efficacy to suppress SPF chickens from shedding challenged virus as in subtype H5N1 inactivated vaccine and to protect SPF chickens against lethal challenge with homologous subtype H5N1 avain influenza virus.The results show that the rFPV-11SH5A has the ability to reduce the adverse effect induced by vaccination stress of FPV.

关 键 词：H5亚型禽流感病毒 血凝素HA基因 重组鸡痘病毒 免疫效力 

分 类 号：S852.65[农业科学—基础兽医学]