CD209L/L-SIGN and CD209/DC-SIGN Act as Receptors for SARS-CoV-2.
CD209L/L-SIGN and CD209/DC-SIGN Act as Receptors for SARS-CoV-2.
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DOI：
10.1101/2020.06.22.165803
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发表时间：
2021-07-28
影响因子：
18.2
通讯作者：
Rahimi, Nader
中科院分区：
文献类型：
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作者：
Amraei, Razie;Yin, Wenqing;Rahimi, Nader
随着COVID-19大流行的持续传播，研究SARS-CoV-2与其宿主之间相互作用的过程具有重要意义。在这里，我们报告了CD209L/L-SIGN和相关蛋白CD209/DC-SIGN作为能够介导SARS-CoV-2进入人细胞的受体的鉴定。人体组织免疫荧光染色显示CD209L在肺、肾上皮和内皮中有显著表达。利用纯化重组SARS-CoV-2刺突受体结合域（S-RBD）或S1同时包含NTB和RBD并异位表达CD209L和CD209进行的多次生化分析显示，CD209L和CD209与S-RBD相互作用。CD209L包含两个n -糖基化序列，位于N92和N361位点，但我们确定只有N92位点被占用。去除该位点的n -糖基化增强了S-RBD与CD209L的结合。CD209L还与ACE2相互作用，提示CD209L和ACE2异源二聚化在SARS-CoV-2进入和感染存在这两种细胞类型中的作用。此外，我们证明了人内皮细胞允许SARS-CoV-2感染，并且通过敲低策略或可溶性CD209L干扰CD209L活性可抑制病毒进入。我们的观察结果表明，CD209L和CD209在包括血管系统在内的疾病相关细胞类型中作为SARS-CoV-2的替代受体。这一特性在ACE2低表达或缺失的组织中尤为重要，可能对抗病毒药物的开发有影响。随着COVID-19大流行的持续传播，研究SARS-CoV-2与其宿主之间相互作用的过程具有重要意义。在这里，我们报告了CD209L/L-SIGN和相关蛋白CD209/DC-SIGN作为能够介导SARS-CoV-2进入人细胞的受体的鉴定。人体组织免疫荧光染色显示CD209L在肺、肾上皮和内皮中有显著表达。利用纯化重组SARS-CoV-2刺突受体结合域（S-RBD）或包含N端结构域和RBD并异位表达CD209L和CD209的S1进行的多次生化分析显示，CD209L和CD209与S-RBD相互作用。CD209L包含两个n -糖基化序列，位于N92和N361位点，但我们确定只有N92位点被占用。去除该位点的n -糖基化增强了S-RBD与CD209L的结合。CD209L还与ACE2相互作用，提示CD209L和ACE2异源二聚化在SARS-CoV-2进入和感染存在这两种细胞类型中的作用。此外，我们证明人类内皮细胞允许SARS-CoV-2感染，并且通过敲低策略或可溶性CD209L干扰CD209L活性可抑制病毒进入。我们的观察结果表明，CD209L和CD209在包括血管系统在内的疾病相关细胞类型中作为SARS-CoV-2的替代受体。这一特性在ACE2低表达或缺失的组织中尤为重要，可能对抗病毒药物的开发有影响。
As the COVID-19 pandemic continues to spread, investigating the processes underlying the interactions between SARS-CoV-2 and its hosts is of high importance. Here, we report the identification of CD209L/L-SIGN and the related protein CD209/DC-SIGN as receptors capable of mediating SARS-CoV-2 entry into human cells. Immunofluorescence staining of human tissues revealed prominent expression of CD209L in the lung and kidney epithelium and endothelium. Multiple biochemical assays using a purified recombinant SARS-CoV-2 spike receptor binding domain (S-RBD) or S1 encompassing both NTB and RBD and ectopically expressed CD209L and CD209 revealed that CD209L and CD209 interact with S-RBD. CD209L contains two N-glycosylation sequons, at sites N92 and N361, but we determined that only site N92 is occupied. Removal of the N-glycosylation at this site enhances the binding of S-RBD with CD209L. CD209L also interacts with ACE2, suggesting a role for heterodimerization of CD209L and ACE2 in SARS-CoV-2 entry and infection in cell types where both are present. Furthermore, we demonstrate that human endothelial cells are permissive to SARS-CoV-2 infection and interference with CD209L activity by knockdown strategy or with soluble CD209L inhibits virus entry. Our observations demonstrate that CD209L and CD209 serve as alternative receptors for SARS-CoV-2 in disease-relevant cell types, including the vascular system. This property is particularly important in tissues where ACE2 has low expression or is absent, and may have implications for antiviral drug development.As the COVID-19 pandemic continues to spread, investigating the processes underlying the interactions between SARS-CoV-2 and its hosts is of high importance. Here, we report the identification of CD209L/L-SIGN and the related protein CD209/DC-SIGN as receptors capable of mediating SARS-CoV-2 entry into human cells. Immunofluorescence staining of human tissues revealed prominent expression of CD209L in the lung and kidney epithelia and endothelia. Multiple biochemical assays using a purified recombinant SARS-CoV-2 spike receptor-binding domain (S-RBD) or S1 encompassing both N termal domain and RBD and ectopically expressed CD209L and CD209 revealed that CD209L and CD209 interact with S-RBD. CD209L contains two N-glycosylation sequons, at sites N92 and N361, but we determined that only site N92 is occupied. Removal of the N-glycosylation at this site enhances the binding of S-RBD with CD209L. CD209L also interacts with ACE2, suggesting a role for heterodimerization of CD209L and ACE2 in SARS-CoV-2 entry and infection in cell types where both are present. Furthermore, we demonstrate that human endothelial cells are permissive to SARS-CoV-2 infection, and interference with CD209L activity by a knockdown strategy or with soluble CD209L inhibits virus entry. Our observations demonstrate that CD209L and CD209 serve as alternative receptors for SARS-CoV-2 in disease-relevant cell types, including the vascular system. This property is particularly important in tissues where ACE2 has low expression or is absent and may have implications for antiviral drug development.