Characterization of CCoV-HuPn-2018 spike protein-mediated viral entry

作者：Yongmei Liu, Danying Chen, Yuanyuan Wang, Xinglin Li, Yaruo Qiu, Mei Zheng, Yanjun Song, Guoli Li, Chuan Song, Tingting Liu, Yuanyuan Zhang, Ju‐Tao Guo, Hanxin Lin, Xuesen Zhao · 发表于：Journal of Virology · 年份：2023 · DOI：10.1128/jvi.00601-23 · 被引用次数：13 · 研究领域：SARS-CoV-2 and COVID-19 Research、Animal Virus Infections Studies、Virus-based gene therapy research

Canine coronavirus-human pneumonia-2018 (CCoV-HuPn-2018) was recently isolated from a child with pneumonia. This novel human pathogen resulted from cross-species transmission of a canine coronavirus. It has been known that CCoV-HuPn-2018 uses aminopeptidase N (APN) from canines, felines, and porcines, but not humans, as functional receptors for cell entry. The molecular mechanism of cell entry in CCoV-HuPn-2018 remains poorly understood. In this study, we demonstrated that among the nine APN orthologs tested, the APN of the Mexican free-tailed bat could also efficiently support CCoV-HuPn-2018 spike (S) protein-mediated entry, raising the possibility that bats may also be an alternative host epidemiologically important for the transmission of this virus. The glycosylation at residue N747 of canine APN is critical for its receptor activity. The gain of glycosylation at the corresponding residues in human and rabbit APNs converted them to functional receptors for CCoV-HuPn-2018. Interestingly, the CCoV-HuPn-2018 spike protein pseudotyped virus infected multiple human cancer cell lines in a human APN-independent manner, whereas sialic acid appeared to facilitate the entry of the pseudotyped virus into human cancer cells. Moreover, while host cell surface proteases trypsin and TMPRSS2 did not promote the entry of CCoV-HuPn-2018, endosomal proteases cathepsin L and B are required for the entry of CCoV-HuPn-2018 in a pH-dependent manner. IFITMs and LY6E are host restriction factors ...