Discovery of novel human and animal cells infected by the severe acute respiratory syndrome coronavirus by replication-specific multiplex reverse transcription-PCR
Discovery of novel human and animal cells infected by the severe acute respiratory syndrome coronavirus by replication-specific multiplex reverse transcription-PCR
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DOI：
10.1128/jcm.42.7.3196-3206.2004
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发表时间：
2004-07-01
影响因子：
9.4
通讯作者：
Wentworth, DE
中科院分区：
文献类型：
--
作者：
Gillim-Ross, L;Taylor, J;Wentworth, DE
严重急性呼吸综合征冠状病毒（SARS-CoV）是最近爆发的严重急性呼吸综合征的病原体。已知对SARS-CoV易感的细胞仅有VeroE6细胞、胎儿恒河猴肾细胞和人外周血单个核细胞。我们开发了一种多重逆转录- pcr方法来分析来自多种组织和物种的细胞对SARS-CoV的易感性。此外，通过细胞上清液的滴定来确定生产性感染。来自三种猴子的细胞对sars冠状病毒敏感。然而，产生的SARS-CoV水平相差4 log（10）。水貂肺上皮细胞（Mv1Lu）和R-Mix（人肺源性细胞（A549）和水貂肺源性细胞（Mv1Lu）的混合单层）被诊断实验室用于检测呼吸道病毒（如流感病毒）；他们还感染了sars冠状病毒，这表明应审查诊断实验室的做法，以确保采取适当的生物安全预防措施。与VeroE6细胞相比，Mv1Lu细胞产生的SARS-CoV很少，这表明它们是一种更安全的SARS-CoV诊断方法。对其他冠状病毒允许的细胞的评估表明，这些细胞类型未被sars冠状病毒感染，这提供了sars冠状病毒结合替代受体的额外证据。对来自肺、肾、肝和肠的人类细胞的分析发现，人类细胞系可被sars冠状病毒有效感染。本研究确定了可用于sars冠状病毒诊断和/或基础研究的新细胞系。我们的数据和其他体内研究表明，SARS-CoV具有广泛的宿主范围，这表明SARS-CoV利用的细胞受体高度保守，并在多种组织中表达。
The severe acute respiratory syndrome coronavirus (SARS-CoV) is the causative agent of the recent outbreak of severe acute respiratory syndrome. VeroE6 cells, fetal rhesus monkey kidney cells, and human peripheral blood mononuclear cells were the only cells known to be susceptible to SARS-CoV. We developed a multiplex reverse transcription-PCR assay to analyze the susceptibility of cells derived from a variety of tissues and species to SARS-CoV. Additionally, productive infection was determined by titration of cellular supernatants. Cells derived from three species of monkey were susceptible to SARS-CoV. However, the levels of SARS-CoV produced differed by 4 log(10). Mink lung epithelial cells (Mv1Lu) and R-Mix, a mixed monolayer of human lung-derived cells (A549) and mink lung-derived cells (Mv1Lu), are used by diagnostic laboratories to detect respiratory viruses (e.g., influenza virus); they were also infected with SARS-CoV, indicating that the practices of diagnostic laboratories should be examined to ensure appropriate biosafety precautions. Mv1Lu cells produce little SARS-CoV compared to that produced by VeroE6 cells, which indicates that they are a safer alternative for SARS-CoV diagnostics. Evaluation of cells permissive to other coronaviruses indicated that these cell types are not infected by SARS-CoV, providing additional evidence that SARS-CoV binds an alternative receptor. Analysis of human cells derived from lung, kidney, liver, and intestine led to the discovery that human cell lines were productively infected by SARS-CoV. This study identifies new cell lines that may be used for SARS-CoV diagnostics and/or basic research. Our data and other in vivo studies indicate that SARS-CoV has a wide host range, suggesting that the cellular receptor(s) utilized by SARS-CoV is highly conserved and is expressed by a variety of tissues.