PDF(Pubmed)
Abstract：
Japanese encephalitis virus (JEV), a flavivirus transmitted by mosquitoes, has caused epidemics and severe neurological diseases in Asian countries. In this study, we developed a cDNA infectious clone, pBAC JYJEV3, of the JEV genotype 3 strain (EF571853.1) using a bacterial artificial chromosome (BAC) vector. The constructed infectious clone was transfected into Vero cells, where it exhibited infectivity and induced cytopathic effects akin to those of the parent virus. Confocal microscopy confirmed the expression of the JEV envelope protein. Comparative analysis of growth kinetics revealed similar replication dynamics between the parental and recombinant viruses, with peak titers observed 72 h post-infection (hpi). Furthermore, plaque assays demonstrated comparable plaque sizes and morphologies between the viruses. Cryo-electron microscopy confirmed the production of recombinant virus particles with a morphology identical to that of the parent virus. Immunization studies in mice using inactivated parental and recombinant viruses revealed robust IgG responses, with neutralizing antibody production increasing over time. These results showcase the successful generation and characterization of a recombinant JEV3 virus and provide a platform for further investigations into JEV pathogenesis and vaccine development.
摘要：
日本脑炎病毒（JEV），由蚊子传播的黄病毒,在亚洲国家引起了流行病和严重的神经系统疾病。在这项研究中,我们开发了一个cDNA感染性克隆，使用细菌人工染色体(BAC)载体的JEV基因型3菌株(EF571853.1)的pBACJYJEV3。将构建的感染性克隆转染到Vero细胞中，它表现出类似于亲本病毒的感染性和诱导的细胞病变效应。共聚焦显微镜证实了JEV包膜蛋白的表达。生长动力学的比较分析揭示了亲本和重组病毒之间相似的复制动力学，感染后72小时观察到峰值滴度（HPI）。此外，噬菌斑试验证明了病毒之间具有可比性的噬菌斑大小和形态。低温电子显微镜证实产生了与亲本病毒相同形态的重组病毒颗粒。使用灭活的亲本和重组病毒在小鼠中进行的免疫研究显示出强大的IgG反应，中和抗体产量随时间增加。这些结果展示了重组JEV3病毒的成功产生和表征，并为进一步研究JEV发病机理和疫苗开发提供了平台。
