表达H9亚型禽流感病毒血凝素基因的重组鸡痘病毒及其免疫效力  

Recombinant Fowlpox Virus Expressing HA From Subtype H9N2 of Avian Influenza Virus and Its Protective Immunity Against Homologous Challenge in Chickens

作　　者：程坚[1] 刘秀梵[1] 彭大新[1] 刘红旗[1] 

机构地区：[1]扬州大学畜禽传染病学农业部重点开放实验室,扬州225009

出　　处：《微生物学报》2002年第4期442-447,共6页Acta Microbiologica Sinica

基　　金："8 6 3"计划资助项目 ( 2 0 0 1AA2 1 30 4 1 )~~

摘　　要：以RT PCR法扩增获得H9亚型禽流感病毒 (AIV)分离株 (A Chicken China F 1 998)的血凝素 (HA)基因 ,将其定向插入鸡痘病毒转移载体 1 1 75的痘苗病毒启动子P7 5的下游 ,得到重组转移载体 1 1 75HA。以脂质体转染法将 1 1 75HA转染至已感染鸡痘病毒 2 82E4疫苗株(wt FPV)的鸡胚成纤维细胞 (CEF)中 ,通过在含X gal的营养琼脂上连续挑选蓝色病毒蚀斑获得并纯化rFPV HA。以间接免疫荧光法证实感染rFPV HA的CEF表达了HA。rFPV HA在免疫 7日龄SPF鸡 7天后即能诱生可检出的血凝抑制 (HI)抗体 ,1 4天后诱生的HI抗体到达高峰 ,且诱生的HI抗体保持较高水平达 5 5天。在 7日龄SPF鸡及含抗FPV母源抗体的商品鸡上进行的免疫效力试验表明 ,rFPV HV能显著抑制静脉攻毒后免疫鸡从泄殖腔的排毒 ,效果与AIV全病毒灭活苗相当。The hemagglutinin (HA) gene from the AIV,A/Chicken/China/F/1998(H9N2),was amplified with the RT PCR technique and directionally inserted into transferring vector 1175,resulted in recombinant transferring vector 1175HA.In order to generate recombinant fowlpox virus expressing HA(rFPV HA),the recombinant transferring vector 1175HA was used to transfect the chicken embryo fibroblasts(CEF)pre infected with wide type fowlpox virus.Then,by selection of blue plaques on the CEF overlaid with agar containing X gal,rFPV HA was obtained and purified.The expression of HA by rFPV HA was detected in the recombinant virus infected CEF by indirect immunofluorescence.Experiments on chickens demonstrated that rFPV HA could induce detectable HI antibodies 7 days post vaccination and those HI antibodies of relatively high titers could persist 55 days.rFPV HA also had the same protective efficacies to suppress SPF chickens or commercial broiler chickens with antibodies against FPV from shedding challenged virus from intestine as inactivated vaccine in oil emulsion.

关 键 词：表达 H9亚型禽流感病毒 血凝素基因 重组鸡痘病毒 免疫效力 

分 类 号：S852.65[农业科学—基础兽医学]