鸡传染性喉气管炎病毒gD基因与ChIL-2重组禽痘病毒的构建及其免疫效力试验  

Construction of recombinant fowl pox virus with ChIL-2 and glycoprotein D of infectious laryngotracheitis virus and its immune efficacy

作　　者：杨明凡[1] 张素梅[1] 陈红英[1] 魏战勇[1] 崔保安[1] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002

出　　处：《中国兽医学报》2012年第12期1768-1772,共5页Chinese Journal of Veterinary Science

基　　金：中国博士后科研基金项目

摘　　要：将鸡传染性喉气管炎病毒(ILTV)gD基因和鸡白细胞介素2(ChIL-2)基因通过同源重组法重组禽痘病毒转移载体,构建含ILTV gD基因和ChIL-2基因的重组禽痘病毒转移载体rFPV-IL2-ILTV-gD,蓝色蚀斑纯化法纯化重组病毒,用IFA检测重组病毒中ILTV gD基因的表达和用ELISA试剂盒检测ChIL-2的表达。重组病毒rFPV-IL2-ILTV-gD免疫试验鸡,间接ELISA测定血清中ILTV抗体效价,动物试验用构建的ILTV强毒以滴鼻方式攻击各组试验鸡,结果表明,外源基因在重组禽痘病毒中得到了稳定表达;ELISA检测结果表明在免疫7 d后能够检测到血清抗体,免疫21 d后血清抗体达到高峰,此后便开始逐渐下降,符合疫苗免疫的消长规律;攻毒后每组鸡的发病情况可以看出重组病毒rFPV-IL2-ILTV-gD组和疫苗对照组对ILTV强毒攻击的保护率分别为96%和92%,而空白对照组为8%。说明构建的重组病毒rFPV-IL2-ILTV-gD免疫效力稍优于弱毒疫苗,能够较好的保护免疫鸡群。The fowl pox transfer vector （pSY681-gD-LacZ） includes the gene of ILTV glycoprotein D and the LacZ gene with promoters in frame of each gene at NotI site. The gene of ChID2 was cloned into pSY538 vector at EcoR I site,then the ChIL-2 gene was cloned into the transfer vector （pSY681-gD-LacZ） with its promoter at Not I site. The recombined transfer vector was transfeeted into the FPV infected chicken embryo fibroblast （CEF） cells by using lipofection procedure. Recombinant virus was selected by blue plague purification and named rFPV-IL2-ILTV-gD. The recombinant virus rFPV-IL2-ILTV-gD, expressing ILTV glycoprotein D was identified by indirect immun- ofluoscenee assay （IFA） and expressing ChIb2 was measured by ELISA. Three groups chickens were immunized with water, commercial ILTV vaccine and recombinant virus rFPV-IL2-ILTV-gD, respectively. Serum antibody of chickens to ILT was measured by ELISA. The chickens were challenged with a lethal dose of virulent ILTV at 28 day post inoculation. Results of immune protection test showed that there was no significant difference between the recombinant virus rFPV-IL2-ILTV-gD and the commercial vaccine and the protection rates of the chickens after the virus challenge were 96 % and 92 %, whereas that of the control group was 8 %. It indicates the recombined virus can effectively protect chicken from ILTV challenge.

关 键 词：鸡传染性喉气管炎病毒gD基因 鸡白细胞介素2基因 重组禽痘病毒 免疫效力 

分 类 号：S852.65[农业科学—基础兽医学]