表达猪圆环病毒2型Cap和猪繁殖与呼吸综合征病毒GP5融合蛋白重组腺病毒的构建和免疫原性测定

Construction and immunogencity of recombinant adenoviruses expressing Cap protein of PCV2 and GP5 protein of PRRSV in mice

摘要:

断奶后多系统衰弱综合征(PMWS)是目前规模化猪场常见的传染病, 由PCV2感染引起。临床上PCV2与PRRSV混合感染较常见, 死亡率和发病率很高, 但目前尚无能有效预防该病的商品化疫苗。本研究利用基因克隆方法, 构建融合表达PCV2-Cap蛋白与PRRSV-GP5蛋白的重组腺病毒rAd-Cap-GP5, 经IPMA、IFA和Western blotting验证了Cap蛋白和GP5蛋白在该重组病毒中获得表达。将该重组腺病毒免疫小鼠测定其抗体、中和抗体, 结果显示小鼠免疫后出现两种病毒的抗体和中和抗体,诱导小鼠产生明显的体液免疫应答, 可以作为预防PCV2-PRRSV混合感染的基因工程疫苗候选毒株, 为PCV2和PRRSV二联重组亚单位疫苗的研究奠定了基础。

Abstract:

Porcine circovirus 2 (PCV2) has been implicated as the etiological agent of postweaning multisystemic wasting syndrome (PMWS). Co-infection of PCV2 and porcine reproductive and respiratory syndrome virus (PRRSV) can result in severe economic losses to the swine industry. In this study, we constructed the recombinant adenovirus rAd-Cap-GP5 expressing Cap of PCV2 and GP5 of PRRSV. And the expression of Cap and GP5 protein in the HEK-293 cells inoculated with rAd-Cap-GP5 were confirmed by immunoperoxidase monolayer assay (IPMA), indirect immunofluorescence assay (IFA) and Western blotting, respectively. The immunogenicity of recombinant adenoviruses rAd-Cap-GP5 was examined in mice by vaccination with the recombinant adenovirus. The results showed that the mice could produce anti-PCV2 and PRRSV antibodies detected by indirect ELISA and virus neutralization assay. It indicated that rAd-Cap-GP5 could provide humoral immunity responses in mice. The recombinant adenovirus rAd-Cap-GP5 might be an attractive candidate vaccine for preventing the disease associated with PCV2 and PRRSV infection.