狂犬病毒CTN株糖蛋白重组鸡痘病毒与核酸疫苗的联合免疫  

Combined immunization of recombinant fowlpox virus containing CTN strain rabies virus glycoprotein gene and nucleic acid vaccine

作　　者：金宁一[1] 郭志儒[1] 赵怀龙[1] 张茂林[1] 

机构地区：[1]解放军军事医学科学院军事兽医研究所,吉林长春130062

出　　处：《免疫学杂志》2004年第4期277-280,共4页Immunological Journal

基　　金：国家自然科学基金项目资助 (30 0 70 568)

摘　　要：目的　构建狂犬病毒糖蛋白重组鸡痘病毒 ,研究联合运用重组病毒与同一抗原核酸疫苗对小鼠的免疫效果 ,为狂犬病疫苗的研究提供基础。方法　以中国鸡痘病毒疫苗株 2 82E4为基础构建的表达载体pUTA 2的ATI·P7.5复合启动子下游 ,插入狂犬病毒CTN 181株糖蛋白基因 ,构建了重组转移载体pUTA RgC。以脂质体转染法将pUTA RgC转染至已感染鸡痘病毒 2 82E4株 (FPV) 2～ 3h的鸡胚成纤维 (CEF)细胞中。待细胞出现明显病变后收获病毒 ,用BrdU进行连续 3次加压筛选 ,挑取蚀斑毒株、纯化、扩增 ,间接免疫荧光鉴定。用重组鸡痘病毒vUTA RgC单独及与核酸疫苗联合免疫小鼠 ,在细胞及体液免疫 ,攻毒保护水平评价所构建的重组病毒及联合免疫效果。结果　间接免疫荧光结果表明已成功构建重组鸡痘病毒 ,免疫后能诱导机体产生细胞与体液免疫 ,并能抵抗标准攻毒株的攻击 ,联合免疫效果较为理想。结论　获得一株能够表达狂犬病毒糖蛋白的vUTA RgC ,并证明有一定的免疫原性 。Objective To construct the rabies virus recombinant fowlpox virus and investigate the combined immunization of the recombinant virus and the nucleic acid vaccine in mice. Methods The fowlpox virus (FPV) transfer vector pUTA-RgC was constructed by inserting the glycoprotein gene of rabies virus CTN-181 strain into the downstream of the hybrid promoter ATI·P7.5 of the FPV expressing vector pUTA-2. The chicken embryo fibroblast (CEF) cells were infected with FPV Chinese vaccine strain 282E4 for 2-3 h. Then the pUTA-RgC was transfected into the CEF cells by lipofectin method. After the appropriate cell pathological effects appeared, the transfected cultures were harvested and passaged for three generations in CEF cells with the MEM cultures containing BrdU. The recombinant FPV was identified by indirect immunofluorescene assay. The recombinant FPV pUTA-RgC and nucleic acid vaccine pVAX-RgC were separately or together injected into the mice. Results The recombinant FPV vUTA-RgC and the recombinant nucleic acid vaccine pVAX-RgC could elicit the cellular and humoral immunity in the mice and afford some protection against the challenge of live rabies virus CVS. Conclusion Combined immunization of recombinant FPV and nucleic acid vaccine can induce potential immune response against rabies virus.

关 键 词：狂犬病毒 重组鸡痘病毒 核酸疫苗 联合免疫 

分 类 号：S852.4[农业科学—基础兽医学]