Vesicular stomatitis virus pseudotyped with severe acute respiratory syndrome coronavirus spike protein
Vesicular stomatitis virus pseudotyped with severe acute respiratory syndrome coronavirus spike protein
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DOI：
10.1099/vir.0.80955-0
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发表时间：
2005-08-01
影响因子：
3.8
通讯作者：
Morikawa, S
中科院分区：
文献类型：
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作者：
Fukushi, S;Mizutani, T;Morikawa, S
严重急性呼吸综合征冠状病毒(SARS-CoV)含有一个单峰蛋白(S)，它与其受体血管紧张素转换酶2(ACE2)结合，诱导膜融合，作为中和抗原。利用水疱性口炎病毒Delta G*系统产生了一株携带水泡性口炎病毒S蛋白的假型病毒。SARS-CoV-S蛋白胞浆结构域的部分缺失使其能够有效地整合到病毒颗粒中，并导致高滴度的假型(VSV-SARS-St19)的产生。VSV-SARS-St19感染Vero E6细胞后，7d即可检测到绿色荧光蛋白的表达。用抗SARS冠状病毒抗体和可溶性ACE2中和VSV-SARS-St19，用抗ACE2抗体处理Vero E6细胞，阻断其感染。这些结果表明，SARS-CoV-S蛋白以ACE2依赖的方式介导VSV-SARS-St19感染。为分析SARS-CoV-S蛋白的功能和建立快速检测SARS-CoV感染中和抗体的方法奠定了基础。
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 VSV Delta G* 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 In 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.