The Spike Protein of the Emerging Betacoronavirus EMC Uses a Novel Coronavirus Receptor for Entry, Can Be Activated by TMPRSS2, and Is Targeted by Neutralizing Antibodies
The Spike Protein of the Emerging Betacoronavirus EMC Uses a Novel Coronavirus Receptor for Entry, Can Be Activated by TMPRSS2, and Is Targeted by Neutralizing Antibodies
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DOI：
10.1128/jvi.00128-13
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发表时间：
2013-05-01
影响因子：
5.4
通讯作者：
Poehlmann, Stefan
中科院分区：
文献类型：
--
作者：
Gierer, Stefanie;Bertram, Stephanie;Poehlmann, Stefan
最近在沙特阿拉伯出现的新型人类冠状病毒EMC(hCoV-EMC)具有高致病性，可能对公众健康构成重大威胁。阐明hCoV-EMC与宿主细胞的相互作用对于我们理解这种病毒的发病机制和确定抗病毒干预的靶点至关重要。在这里，我们研究了控制hCoV-EMC进入宿主细胞的病毒和细胞决定因素。我们发现，人冠状病毒-EMC(EMC-S)的刺突蛋白被掺入慢病毒颗粒中，并介导来自不同器官的人细胞系的转导，包括肺、肾、结肠以及原代人巨噬细胞。已知的冠状病毒受体ACE2、CD13和CEACAM1的表达不促进EMC-S驱动的转导，提示hCoV-EMC使用一种新的受体进入。蛋白水解酶的定向表达和抑制分析表明，TMPRSS2和内体组织蛋白激活了EMC-S的病毒-细胞融合，成为抗病毒干预的潜在靶点。最后，EMC-S驱动的转导被hCoV-EMC感染患者的血清阻断，表明患者可以产生EMC-S特异性的中和抗体。总而言之，我们的结果表明，hCoV-EMC使用一种新的受体来使蛋白酶激活进入人类细胞，并可能能够在肺外传播。此外，他们将TMPRSS2、组织蛋白B和L定义为潜在的干预目标，并提示中和抗体有助于控制hCoV-EMC感染。
The novel human coronavirus EMC (hCoV-EMC), which recently emerged in Saudi Arabia, is highly pathogenic and could pose a significant threat to public health. The elucidation of hCoV-EMC interactions with host cells is critical to our understanding of the pathogenesis of this virus and to the identification of targets for antiviral intervention. Here we investigated the viral and cellular determinants governing hCoV-EMC entry into host cells. We found that the spike protein of hCoV-EMC (EMC-S) is incorporated into lentiviral particles and mediates transduction of human cell lines derived from different organs, including the lungs, kidneys, and colon, as well as primary human macrophages. Expression of the known coronavirus receptors ACE2, CD13, and CEACAM1 did not facilitate EMC-S-driven transduction, suggesting that hCoV-EMC uses a novel receptor for entry. Directed protease expression and inhibition analyses revealed that TMPRSS2 and endosomal cathepsins activate EMC-S for virus-cell fusion and constitute potential targets for antiviral intervention. Finally, EMC-S-driven transduction was abrogated by serum from an hCoV-EMC-infected patient, indicating that EMC-S-specific neutralizing antibodies can be generated in patients. Collectively, our results indicate that hCoV-EMC uses a novel receptor for protease-activated entry into human cells and might be capable of extrapulmonary spread. In addition, they define TMPRSS2 and cathepsins B and L as potential targets for intervention and suggest that neutralizing antibodies contribute to the control of hCoV-EMC infection.