狂犬病病毒糖、核蛋白基因疫苗的构建及在小鼠的免疫试验  

Construction of Rabies Virus Glycoprotein/Nucleoprotein Genetic Vaccines and Immunological Experiment on Mice

作　　者：袁慧君[1] 扈荣良[1] 包世俊[1] 张守峰[1] 殷震[1] 

机构地区：[1]解放军军需大学军事兽医研究所,吉林长春130062

出　　处：《中国兽医学报》2004年第2期141-144,共4页Chinese Journal of Veterinary Science

基　　金：国家"8 63"计划资助项目 (2 0 0 1AA2 13 14 1)

摘　　要：分别以 p IRES1neo和 p VAX1为载体 ,构建了以狂犬病病毒糖蛋白和核蛋白为目的基因的基因疫苗单或双基因表达载体。以司苯 -甘油作为包裹剂 ,按 1μg DNA用 0 .5μL司苯 -甘油比例混合 ,将各表达载体制成 DNA疫苗 ,进行小鼠免疫试验。免疫共分 p IRES1neo、p IG- neo、p IN- neo、p IGN、p IG- neo+p IN- neo、p VAX1、p VG、p VN和 p VGN9个试验组 ,每只小鼠于后肢胫前肌每侧接种基因疫苗 5 0μL (0 .5 g/ L ) ,共免疫 3次 ,间隔 14 d。通过间接 EL ISA和细胞中和指数测定分别检测免疫应答水平。结果显示 ,1免疫 3次后 ,所有试验组抗体阳转率 10 0 % ;2第 3次免疫后 14 d检测 ,抗体水平显著上升 ;3以 p IRES1neo为载体构建的基因疫苗免疫效果普遍优于以 p VAX1为载体构建的基因疫苗 ,其中含有糖蛋白 /核蛋白双基因共表达载体 p IGN免疫组的抗体水平均高于其他试验组 ;4以 7.5个 L D50 的 CVS强毒肌肉攻毒 ,保护率达 89%。通过以上试验证明 ,p IGN是用于本动物免疫试验的最佳 DNA疫苗。Recombinant monogene and double gene expression vectors pIG-neo, pIN-neo, pVG, pVN, pIGN and pVGN were constructed by inserting the G and N genes into pIRES1neo and pVAX1,respectively. Span-glycerol was chosen as adjuvant and mixed with the plasmids to form DNA vaccine. Nine groups of mice were immunized with a dosage of 100 μL(0.5 g/L) of plasmids pIRES1neo, pIG-neo, pIN-neo, pIGN, pIG+pIN, pVAX1, pVG, pVN and pVGN, respectively, for 3 times at 14 days intervals. Indirect ELISA and neutralization were employed to determine the immunological response. Results showed that: (1)After 3 times of immunization, seroconversion occurred in all groups excepting negative controls. (2)Antibody level went up swiftly 14 days after the third injection of DNA vaccine. (3)The immunological effect of DNA vaccines based on the pIRES1neo was overally better than those based on the pVAX1. Of all the vaccines, the double gene expression vector pIGN haboring both the glycoprotein and the nucleoprotein cDNAs was the best. The antibody level it stimulated was significantly higher than the others. (4)The protective rate against the challenge of 7.5 LD_(50) CVS was 89%. It indicated that it was worthy being assayed in dogs.

关 键 词：狂犬病病毒 糖蛋白 核蛋白 基因疫苗 小鼠 免疫 

分 类 号：S852.655[农业科学—基础兽医学] Q78[农业科学—兽医学]