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用编码猪繁殖与呼吸综合征病毒ORF5的质粒DNA接种的猪的免疫反应。

Immune response in pigs vaccinated with plasmid DNA encoding ORF5 of porcine reproductive and respiratory syndrome virus.

作者信息

Pirzadeh B, Dea S

机构信息

Centre de recherche en virologie, Institut Armand-Frappier, Université du Québec, Laval-des-Rapides, Canada.

出版信息

J Gen Virol. 1998 May;79 ( Pt 5):989-99. doi: 10.1099/0022-1317-79-5-989.

Abstract

The ORF5-encoded major envelope glycoprotein (GP5) of porcine reproductive and respiratory syndrome virus (PRRSV) is one of the three major structural proteins of this virus. While some porcine convalescent sera and monoclonal antibodies directed against GP4 and GP5 have the capacity to neutralize the virus in vitro, the protein specificity of porcine neutralizing sera has not yet been established. DNA immunization with a plasmid encoding GP5 of PRRSV, under the control of a human cytomegalovirus promoter, induced anti-GP5-specific neutralizing antibodies in pigs and BALB/c mice. The GP5 protein specificity of neutralizing sera was confirmed by immunoblotting and ELISA. Peripheral blood mononuclear cells obtained from DNA-vaccinated pigs underwent blastogenic transformation in the presence of E. coli-expressed recombinant ORF5-encoded protein, indicating the specificity of the cellular immune response to GP5. Following a massive intratracheal challenge with the virulent IAF-Klop strain of PRRSV, DNA-vaccinated pigs were protected from generalized viraemia and the development of typical macroscopic lung lesions that were observed in unvaccinated, virus-challenged controls, as well as in pigs that were immunized with E. coli-expressed GST-ORF5 recombinant fusion protein. Interstitial pneumonitis and broncho-alveolitis were remarkably milder in DNA-vaccinated animals. These results suggest that the GP5 of PRRSV is a good candidate for a subunit recombinant-type vaccine.

摘要

猪繁殖与呼吸综合征病毒(PRRSV)的开放阅读框5(ORF5)编码的主要包膜糖蛋白(GP5)是该病毒的三种主要结构蛋白之一。虽然一些猪恢复期血清以及针对GP4和GP5的单克隆抗体具有在体外中和该病毒的能力,但猪中和血清的蛋白特异性尚未确定。用人巨细胞病毒启动子控制下的编码PRRSV的GP5的质粒进行DNA免疫,可在猪和BALB/c小鼠中诱导产生抗GP5特异性中和抗体。通过免疫印迹和酶联免疫吸附测定(ELISA)证实了中和血清的GP5蛋白特异性。从经DNA疫苗接种的猪获得的外周血单个核细胞在大肠杆菌表达的重组ORF5编码蛋白存在下发生增殖性转化,表明细胞免疫反应对GP5具有特异性。在用PRRSV的强毒株IAF-Klop进行大规模气管内攻毒后,经DNA疫苗接种的猪免受全身性病毒血症的影响,并且未观察到在未接种疫苗、经病毒攻毒的对照猪以及用大肠杆菌表达的GST-ORF5重组融合蛋白免疫的猪中出现的典型宏观肺部病变。在经DNA疫苗接种的动物中,间质性肺炎和支气管肺泡炎明显较轻。这些结果表明,PRRSV的GP5是亚单位重组型疫苗的良好候选者。

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