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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病毒的构建及免疫原性

[Construction and immunogenicity of recombinant pseudorabies virus expressing the modified GP5m protein of porcine reproduction and respiratory syndrome virus].

作者信息

Jiang Yun-Bo, Fang Liu-Rong, Xiao Shao-Bo, Zhang Hui, Chen Huan-Chun

机构信息

Laboratory of Animal Virology, College of Animal Science and Veterinary Medicine, Huazhong Agriculture University, Wuhan 430070, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2005 Nov;21(6):858-64.

Abstract

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, ORF5m. 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 GP5m protein is a candidate for the development of bivalent genetic engineering vaccines against PRRSV and PRV.

摘要

伪狂犬病病毒(PRV)是一种α疱疹病毒,已被用作活病毒动物疫苗的载体。表达猪繁殖与呼吸综合征病毒(PRRSV)GP5蛋白的重组PRV TK-/gE-/GP5+,是基于PRV基因缺失疫苗病毒株TK-/gE-/LacZ+构建的。然而,该毒株刺激接种动物产生抗PRRSV中和抗体的能力较差。为了增强PRV重组体的特异性免疫应答,用修饰的ORF5基因ORF5m替换了PRRSV TK-/gE-/LacZ+的ORF5基因。通过PCR、Southern印迹和Western印迹对所得重组PRV TK-/gE-/GP5m+进行了验证。将TK-/gE-/GP5m+和表达GP5蛋白的TK-/gE-/GP5+接种到Balb/c小鼠中以评估它们的免疫原性。结果表明,TK-/gE-/GP5m+诱导的抗PRRSV中和抗体量和细胞免疫应答高于TK-/gE-/GP5+。本研究表明,表达修饰的GP5m蛋白的新型重组PRV是开发抗PRRSV和PRV二价基因工程疫苗的候选者。

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