Retroviruses pseudotyped with the severe acute respiratory syndrome coronavirus spike protein efficiently infect cells expressing angiotensin-converting enzyme 2
Retroviruses pseudotyped with the severe acute respiratory syndrome coronavirus spike protein efficiently infect cells expressing angiotensin-converting enzyme 2
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DOI：
10.1128/jvi.78.19.10628-10635.2004
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发表时间：
2004-10-01
影响因子：
5.4
通讯作者：
Choe, H
中科院分区：
文献类型：
--
作者：
Moore, MJ;Dorfman, T;Choe, H
冠状病毒对受体细胞的感染是由其尖峰蛋白(S)介导的。引起严重急性呼吸综合征(SARS)的冠状病毒(SARS-CoV)感染表达受体血管紧张素转换酶2(ACE2)的细胞。在这里，我们发现对SARS冠状病毒S蛋白基因的密码子优化大大提高了S蛋白的表达。我们还发现两种逆转录病毒，猴免疫缺陷病毒和小鼠白血病病毒，都表达绿色荧光蛋白，并与SARS冠状病毒S蛋白或S蛋白变体假型，有效地感染稳定表达血管紧张素转换酶2的HEK293T细胞。在两种情况下，由胞浆结构域被截短并改变为包括人类免疫缺陷病毒1型包膜糖蛋白胞质尾部片段的S蛋白突变体介导的感染都比野生型S蛋白介导的感染效率高得多。利用S蛋白假型SIV，我们发现ACE2的酶活性在S蛋白介导的感染中没有作用。最后，我们证明了一种可溶的和催化失活的ACE2形式有效地阻断了S蛋白假型逆转录病毒和SARS冠状病毒的感染。这些结果使得在不使用活病毒的情况下研究SARS-CoV进入抑制剂成为可能，并建议了一种治疗SARS的候选方法。
Infection of receptor-bearing cells by coronaviruses is mediated by their spike (S) proteins. The coronavirus (SARS-CoV) that causes severe acute respiratory syndrome (SARS) infects cells expressing the receptor angiotensin-converting enzyme 2 (ACE2). Here we show that codon optimization of the SARS-CoV S-protein gene substantially enhanced S-protein expression. We also found that two retroviruses, simian immunodeficiency virus (SIV) and murine leukemia virus, both expressing green fluorescent protein and pseudotyped with SARS-CoV S protein or S-protein variants, efficiently infected HEK293T cells stably expressing ACE2. Infection mediated by an S-protein variant whose cytoplasmic domain had been truncated and altered to include a fragment of the cytoplasmic tail of the human immunodeficiency virus type 1 envelope glycoprotein was, in both cases, substantially more efficient than that mediated by wild-type S protein. Using S-protein-pseudotyped SIV, we found that the enzymatic activity of ACE2 made no contribution to S-protein-mediated infection. Finally, we show that a soluble and catalytically inactive form of ACE2 potently blocked infection by S-protein-pseudotyped retrovirus and by SARS-CoV. These results permit studies of SARS-CoV entry inhibitors without the use of live virus and suggest a candidate therapy for SARS.