Fukushi Shuetsu, Mizutani Tetsuya, Saijo Masayuki, Matsuyama Shutoku, Miyajima Naoko, Taguchi Fumihiro, Itamura Shigeyuki, Kurane Ichiro, Morikawa Shigeru
Special Pathogens Laboratory, Department of Virology I, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashimurayama, Tokyo 208-0011, Japan.
Laboratory of Respiratory Viral Diseases and SARS, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashimurayama, Tokyo 208-0011, Japan.
J Gen Virol. 2005 Aug;86(Pt 8):2269-2274. doi: 10.1099/vir.0.80955-0.
Severe acute respiratory syndrome coronavirus (SARS-CoV) contains a single spike (S) protein, which binds to its receptor, angiotensin-converting enzyme 2 (ACE2), induces membrane fusion and serves as a neutralizing antigen. A SARS-CoV-S protein-bearing vesicular stomatitis virus (VSV) pseudotype using the VSVDeltaG* system was generated. Partial deletion of the SARS-CoV-S protein cytoplasmic domain allowed efficient incorporation into VSV particles and led to the generation of a pseudotype (VSV-SARS-St19) at high titre. Green fluorescent protein expression was demonstrated as early as 7 h after infection of Vero E6 cells with VSV-SARS-St19. VSV-SARS-St19 was neutralized by anti-SARS-CoV antibody and soluble ACE2, and its infection was blocked by treatment of Vero E6 cells with anti-ACE2 antibody. These results indicated that VSV-SARS-St19 infection is mediated by SARS-CoV-S protein in an ACE2-dependent manner. VSV-SARS-St19 will be useful for analysing the function of SARS-CoV-S protein and for developing rapid methods of detecting neutralizing antibodies specific for SARS-CoV infection.
严重急性呼吸综合征冠状病毒（SARS-CoV）含有单一的刺突（S）蛋白，该蛋白与其受体血管紧张素转换酶2（ACE2）结合，诱导膜融合并作为中和抗原。利用VSVDeltaG*系统构建了携带SARS-CoV-S蛋白的水疱性口炎病毒（VSV）假型。SARS-CoV-S蛋白胞质结构域的部分缺失允许其高效掺入VSV颗粒，并导致高滴度假型（VSV-SARS-St19）的产生。在用VSV-SARS-St19感染Vero E6细胞后7小时，即可检测到绿色荧光蛋白表达。VSV-SARS-St19被抗SARS-CoV抗体和可溶性ACE2中和，其感染可通过用抗ACE2抗体处理Vero E6细胞来阻断。这些结果表明，VSV-SARS-St19感染是以ACE2依赖的方式由SARS-CoV-S蛋白介导的。VSV-SARS-St19将有助于分析SARS-CoV-S蛋白的功能，并有助于开发快速检测针对SARS-CoV感染的中和抗体的方法。
