狂犬病病毒囊膜糖蛋白基因DNA免疫研究  

Immunogenicity of a codon optimized DNA vaccine expressing rabies virus G protein

作　　者：蒋伟[1,2] 王喜军[2] 帅磊[2] 葛金英[2] 任家琰[1] 步志高[2] 

机构地区：[1]山西农业大学动物科技学院,山西太谷030801 [2]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2013年第9期752-755,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家科技重大专项(2012ZX10004214)

摘　　要：狂犬病病毒(RV)囊膜糖蛋白(G)是该病毒诱导保护性免疫反应的主要抗原。为研究G蛋白基因的DNA免疫效果,本研究构建了表达哺乳动物密码子优化的RV G基因的重组真核表达质粒pCAGG-RVG,间接免疫荧光试验及western blot结果表明,重组RV G基因在pCAGG-RVG转染的BHK-21细胞中获得正确表达。将pCAGG-RVG按500μg剂量经肌肉注射途径免疫比格犬,间隔4周以相同剂量、途径加强免疫一次,并于初次免疫前、后不同时间检测血清RV的中和抗体。结果显示,pCAGG-RVG一次免疫即可诱导产生显著的中和抗体反应,二次免疫中和抗体滴度表现出显著地增强效果。二次免疫后,中和抗体滴度缓慢下降,保持持续平稳,至初次免疫后第54周,7只免疫犬病毒中和抗体滴度平均为2.88 IU/mL,并且全部高于0.5 IU/mL,即全部高于对强毒攻击保护的最低中和抗体滴度要求。因此,本研究构建的pCAGG-RVG具有预防狂犬病的潜力,是一种有希望的狂犬病候选疫苗。The glycoprotein （G） of rabies virus （RV） is a major antigen for protective immune responses against RV infection. In this study, G gene of RV was optimized according to mammalian favored codons and cloned into eukaryotic expression vector pCAGGS to construct recombinant plasmid pCAGG-RVG, and the G protein was expressed transiently in BHK-21 cells identified by indirect immunofluorescence assay and western bolt. In addition, Beagle dogs were immunized with 500 μg of pCAGG-RVG via intramuscular inoculation for primary and booster vaccination at 4 weeks interval. The results of virus neutralization demonstrated that the dogs developed the neutralization antibody （NA） at average of 3.81 IU/mL to RV post the initial vaccination, and the NA titers reached a pike level at average of 8.21 IU/mL in 2 weeks post the booster. Thereafter, the NA in the dogs maintained at high titre （2.88 IU/mL） during 54 weeks of the experimental period, which exceed the protection threshold value of 0.5 IU/mL of the minimum NA titer required for protecting animals from challenge with street RV. Therefore, the recombinant plasmid pCAGG-RVG has the potential to serve as a DNA vaccine against rabies in dogs.

关 键 词：狂犬病病毒 DNA疫苗 G蛋白 

分 类 号：S852.65[农业科学—基础兽医学]