表达PRRSV GP2a-GP4的重组LCMV的构建与鉴定

Establishment and Identification of A Recombinant LCMV Expressing PRRSV GP2a-GP4

猪繁殖与呼吸综合征病毒(Porcine reproductive and respiratory syndrome virus, PRRSV)是危害养猪业发展的重要病原之一。高效PRRS疫苗的研发是病毒防控的重要前提。借助机体产生针对病毒颗粒表面糖蛋白(glycoprotein, GP)GP2a/GP4的中和抗体,阻止与CD163受体的结合,成为PRRSV疫苗研发的关键策略之一。据此,构建了基于重组淋巴细胞性脉络丛脑膜炎病毒(Lymphocytic choriomeningitis virus, LCMV)的反向遗传学操作系统,使重组LCMV携带PRRSV GP2a-GP4蛋白的基因序列。对比P2代和P16代重组病毒的生长曲线发现P16代病毒增殖速度更快、滴度更高。在重组LCMV感染的细胞中,通过Western blot试验能够检测到GP2a-GP4蛋白的表达。该重组LCMV载体的免疫保护效果有待攻毒保护试验的进一步评估。

Porcine reproductive and respiratory syndrome virus(PRRSV)has plagued the swine industry for 30 years and the development of effective vaccines would be an important step towards the control of PRRS.One potential strategy is the induction of antibody that disrupts the interaction between the porcine CD163 and the GP2a-GP4 from the PRRSV virion.This study described the successful development of reverse genetics systems to generate a recombinant lymphocytic choriomeningitis virus(LCMV)expressing PRRSV GP2a-GP4 antigens.Compared the viral growth curves between recombinant LCMV P2 and P16 found that P16 virus reached higher titer.The expression of GP2a-GP4 protein was verified by Western blot in recombinant LCMV-infected cells.Future PRRSV challenge experiments in swine will be needed to assess immune protection by this recombinant LCMV vector.