中国兽医科学2011，Vol.41Issue(1)：53-59,7.
PRRSV GP3蛋白和PCV-2Cap蛋白在杆状病毒囊膜表面的共展示及展示蛋白的小鼠免疫试验
Baculovirus surface display of GP3 envelope glycoprotein of PRRSV and Cap protein of PCV-2 and immunogenicity of the displayed proteins in mouse model
摘要
Abstract
In order to develope divalent genetic engineering subunit vaccines against both porcine reproductive and respiratory syndrome(PRRS) and post-weaning multisystemic wasting syndrome(PMWS)based on baculovirus surface display system, one recombinant baculovirus BacSC-Dual-ORF3-ORF2 expressing both histidine-tagged PRRSV GP3 protein and porcine circovirus type 2(PCV-2) Cap protein with the CTD and TM derived from baculovirus envelope protein gp64 was constructed. The recombinant baculovirus was analyzed by Western-blot,immunofluorescence confocal microscopy,immunogold electron microscopy, animal immune experiments serum neutralization test and lymphocyte proliferative responses. After infection,GP3 and Cap proteins were expressed and anchored on the plasma membrane of Sf-9 cells, revealed by confocal microscopy. The results of immunogold electron microscope demonstrated that the Bac-SC-Dual-ORF3-ORF2 displayed GP3 and Cap proteins on the viral surface. The animal immune experiments showed that BacSC-Dual-ORF3-ORF2 elicited high antibody of PRRSV and PCV-2 in the treated mouse models. The serum neutralization test showed that the serum from the BacSC-Dual-ORF3-ORF2 treated models had significant levels of virus neutralization activities and neutralizing antibody titer was 1 : 9.6 (PRRSV) and 1: 13.6 (PCV-2), respectively. Lymphocyte proliferative responses results indicated that recombinant BacSC-Dual-ORF3-ORF2 also induced an obvious cellular immune response. The recombinant baculovirus BacSC-Dual-ORF3-ORF2 displaying GP3 and Cap protein was successfully developed. The results showed that the recombinant baculovirus was a potential bivalent genetic engineering marker vaccine against both PRRSV and PCV-2.关键词
杆状病毒表面展示系统/猪繁殖与呼吸综合征病毒/猪圆环病毒2型/GP3蛋白/Cap蛋白分类
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许信刚,张宽,王志异,童德文..PRRSV GP3蛋白和PCV-2Cap蛋白在杆状病毒囊膜表面的共展示及展示蛋白的小鼠免疫试验[J].中国兽医科学,2011,41(1):53-59,7.基金项目
家畜疫病病原生物学国家重点实验室开放基金课题(SKLVEB2009KFKT002) （SKLVEB2009KFKT002）
教育部新世纪优秀人才支持计划项目(NCET-07-0701) （NCET-07-0701）
