PDF(Pubmed)
Abstract：
Rabies virus (RABV) is the causative agent of rabies, a lethal neurological disease in mammals. RABV strains can be classified into fixed strains (laboratory strains) and street strains (field/clinical strains), which have different properties including cell tropism and neuroinvasiveness. RABV Toyohashi strain is a street strain isolated in Japan from an imported case which had been bitten by rabid dog in the Philippines. In order to facilitate molecular studies of RABV, we established a reverse genetics (RG) system for the study of the Toyohashi strain. The recombinant virus was obtained from a cDNA clone of Toyohashi strain and exhibited similar growth efficiency as the original virus in cultured cell lines. Both the original and recombinant strains showed similar pathogenicity with high neuroinvasiveness in mice, and the infected mice developed a long and inconsistent incubation period, which is characteristic of street strains. We also generated a recombinant Toyohashi strain expressing viral phosphoprotein (P protein) fused with the fluorescent protein mCherry, and tracked the intracellular dynamics of the viral P protein using live-cell imaging. The presented reverse genetics system for Toyohashi strain will be a useful tool to explore the fundamental molecular mechanisms of the replication of RABV street strains.
摘要：
狂犬病病毒（RABV）是狂犬病的病原体，哺乳动物的致命神经系统疾病。RABV菌株可分为固定菌株（实验室菌株）和街道菌株（现场/临床菌株），它们具有不同的特性，包括细胞嗜性和神经侵袭性。RABVToyohashi毒株是日本的街头毒株，从菲律宾的狂犬病狗咬伤的进口病例中分离出来。为了促进RABV的分子研究，我们建立了用于研究丰桥菌株的反向遗传学（RG）系统。重组病毒是从Toyohashi株的cDNA克隆获得的，在培养的细胞系中表现出与原始病毒相似的生长效率。原始菌株和重组菌株在小鼠中显示出相似的致病性，具有高神经侵袭性，受感染的小鼠发展了漫长而不一致的潜伏期，这是街头菌株的特征。我们还产生了表达与荧光蛋白mCherry融合的病毒磷蛋白（P蛋白）的重组Toyohashi菌株，并使用活细胞成像跟踪病毒P蛋白的细胞内动力学。提出的Toyohashi菌株的反向遗传系统将是探索RABV街道菌株复制的基本分子机制的有用工具。
