表达猪繁殖与呼吸综合征病毒修饰型GP5m蛋白的重组伪狂犬病毒的构建及其在小鼠的免疫反应  

Construction and Immunogenicity of Recombinant Pseudorabies Virus Expressing the Modified GP5m Protein of Porcine Reproduction and Respiratory Syndrome Virus

作　　者：江云波[1] 方六荣[1] 肖少波[1] 张辉[1] 陈焕春[1] 

机构地区：[1]华中农业大学动物医学院动物病毒研究室,武汉430070

出　　处：《生物工程学报》2005年第6期858-864,共7页Chinese Journal of Biotechnology

基　　金：国家自然科学基金(No.30300257);国家基础研究973计划(No.2005CB523200);武汉市青年科技晨光计划(No.20025001041)资助项目~~

摘　　要：猪伪狂犬病毒(PRV)是一种良好的兽用活病毒疫苗载体。但以PRV基因缺失疫苗株TK-gE-LacZ+为载体表达PRRSVGP5的重组病毒TK-gE-GP5+免疫实验动物后难以激发抗PRRSV的中和抗体。为了进一步增强这种重组病毒的免疫效力,用具有更好免疫原性的修饰的ORF5基因(ORF5m)代替天然ORF5基因,构建了表达PRRSV的修饰型GP5m蛋白的重组伪狂犬病毒TK-gE-GP5m+。经PCR、Southernblot、Westernblot证实构建正确,并能表达具有活性的GP5m蛋白。将TK-gE-GP5m+与TK-gE-GP5+分别免疫Balbc小鼠,结果TK-gE-GP5m+免疫小鼠不仅产生了较高水平的抗PRRSV的中和抗体(36只达到了1∶16),而且在诱导PRRSV特异性细胞免疫方面也显著优于TK-gE-GP5+,表明TK-gE-GP5m+是一种极有希望的PRRSV和PRV二价基因工程候选疫苗。Pseudorabies virus （PRV）, an alpha-herpesvirus, has been used as a vector for live-viral animal vaccines. The recombinant PRV TK^-gE^-/GP5^＋ , which expressing GP5 of PRRSV, is developed based on the PRV genetic-depleting vaccine-virus strain, TK^-/gE^-/LacZ^＋ . However, this strain stimulated poorly the vaccinated animals to produce neutralizing antibodies against PRRSV. In order to develop a booster specific immunized response of the PRV recombinant, the ORF5 gene of PRRSV TK^-/gE^-/LacZ^＋ was substituted by a modified ORF5 gene, ORFSm, The resultant recombinant PRV, TK^-gE^-/GP5m^＋ , was verified by PCR, Southern blotting and Western blotting, TK^-gE^-/GP5m^＋ and TK^-gE^-/GP5^＋ expressed GP5 proteins were inoculated into balb/c mice to evaluate their immunogenicity, The results demonstrated that the amount of neutralization antibodies and cell-immunity responses induced by TK^-gE^-/GP5m^＋ against PRRSV were higher than that of TK^-gE^-/GP5^＋ , This study indicated that the new recombinant PRV expressing the modified GPSm protein is a candidate for the development of bivalent genetic engineering vaccines against PRRSV and PRV.

关 键 词：猪繁殖与呼吸综合征病毒 GP5m蛋白 重组伪狂犬病毒 

分 类 号：S858.28[农业科学—临床兽医学]