Severe acute respiratory syndrome coronavirus infection of human ciliated airway epithelia: Role of ciliated cells in viral spread in the conducting airways of the lungs
Severe acute respiratory syndrome coronavirus infection of human ciliated airway epithelia: Role of ciliated cells in viral spread in the conducting airways of the lungs
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DOI：
10.1128/jvi.79.24.15511-15524.2005
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发表时间：
2005-12-01
影响因子：
5.4
通讯作者：
Pickles, RJ
中科院分区：
文献类型：
--
作者：
Sims, AC;Baric, RS;Pickles, RJ
严重急性呼吸综合征冠状病毒（SARS-CoV）于2002年出现，是人类严重下呼吸道感染的重要病因，需要体外肺模型来阐明病毒感染的细胞靶点和后果。SARS-CoV受体人血管紧张素i转换酶2 （hACE2）在来源于鼻或气管区的人气道组织的纤毛气道上皮细胞中检测到，提示SARS-CoV可能感染近端气道。为了评估来自鼻腔和气管支气管气道区域的人纤毛气道上皮（HAE）体外模型的感染性，我们通过删除开放阅读框7a/7b （ORF7a/ 7b）并插入绿色荧光蛋白（GFP）产生重组SARS-CoV，从而获得SARS-CoV GFP。SARS-CoV GFP在细胞系中复制到与野生型病毒相似的滴度。sars冠状病毒通过根尖表面特异性感染HAE，并在感染后48小时复制到107 PFU/ml滴度。针对hACE2的多克隆抗血清阻断了病毒感染和复制，提示hACE2是SARS-CoV感染HAE的主要受体。定位于纤毛上皮细胞的sars冠状病毒结构蛋白和病毒粒子。感染是高度细胞溶解性的，因为感染的纤毛细胞坏死并随时间脱落到上皮的管腔表面。sars冠状病毒GFP在来自仓鼠或恒河猴呼吸道的纤毛细胞培养物中也有较小程度的复制。SARS-CoV对HAE患者纤毛细胞的有效感染为研究SARS-CoV复制和发病机制的特点提供了一个有用的体外人肺源模型。
Severe acute respiratory syndrome coronavirus (SARS-CoV) emerged in 2002 as an important cause of severe lower respiratory tract infection in humans, and in vitro models of the lung are needed to elucidate cellular targets and the consequences of viral infection. The SARS-CoV receptor, human angiotensin I-converting enzyme 2 (hACE2), was detected in ciliated airway epithelial cells of human airway tissues derived from nasal or trachcobronchial regions, suggesting that SARS-CoV may infect the proximal airways. To assess infectivity in an in vitro model of human ciliated airway epithelia (HAE) derived from nasal and tracheobronchial airway regions, we generated recombinant SARS-CoV by deletion of open reading frame 7a/7b (ORF7a/ 7b) and insertion of the green fluorescent protein (GFP), resulting in SARS-CoV GFP. SARS-CoV GFP replicated to titers similar to those of wild-type viruses in cell lines. SARS-CoV specifically infected HAE via the apical surface and replicated to titers of 107 PFU/ml by 48 h postinfection. Polyclonal antisera directed against hACE2 blocked virus infection and replication, suggesting that hACE2 is the primary receptor for SARS-CoV infection of HAE. SARS-CoV structural proteins and virions localized to ciliated epithelial cells. Infection was highly cytolytic, as infected ciliated cells were necrotic and shed over time onto the luminal surface of the epithelium. SARS-CoV GFP also replicated to a lesser extent in ciliated cell cultures derived from hamster or rhesus monkey airways. Efficient SARS-CoV infection of ciliated cells in HAE provides a useful in vitro model of human lung origin to study characteristics of SARS-CoV replication and pathogenesis.