Wild-type human coronaviruses prefer cell-surface TMPRSS2 to endosomal cathepsins for cell entry

• Published in: Virology (New York, N.Y.) Vol. 517; pp. 9 - 15
• Main Authors: Shirato, Kazuya, Kawase, Miyuki, Matsuyama, Shutoku
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.04.2018
• Subjects: Air-liquid interface culture; Amino Acid Sequence; animal viruses; Betacoronavirus 1; Brief Communication; cathepsin L; Cathepsin L - genetics; Cathepsins; Cell Line; Coronavirus - physiology; cultured cells; Endosomes; Entry; epithelial cells; host-pathogen relationships; Human bronchial tracheal epithelial cells; Human coronavirus; Human coronavirus 229E; Human coronavirus HKU1; Humans; pathogenesis; RNA, Messenger - genetics; RNA, Messenger - metabolism; serine; Serine Endopeptidases - physiology; serine proteinases; Spike Glycoprotein, Coronavirus; Virus Internalization; Virus Replication - physiology; DMEM; Entry; HCoV; S; Air-liquid interface culture; CatL; VHCR; Human bronchial tracheal epithelial cells; ATCC; Cam; Human coronavirus
• DOI: 10.1016/j.virol.2017.11.012
• ISSN: 0042-6822; 1096-0341

Human coronaviruses (HCoVs) enter cells via two distinct pathways: the endosomal pathway using cathepsins to activate spike protein and the cell-surface or early endosome pathway using extracellular proteases such as transmembrane protease serine 2 (TMPRSS2). We previously reported that clinical isolates of HCoV-229E preferred cell-surface TMPRSS2 to endosomal cathepsin for cell entry, and that they acquired the ability to use cathepsin L by repeated passage in cultured cells and were then able to enter cells via the endosomal pathway. Here, we show that clinical isolates of HCoV-OC43 and -HKU1 preferred the cell-surface TMRRSS2 to endosomal cathepsins for cell entry, similar to HCoV-229E. In addition, the cell-culture-adapted HCoV-OC43 lost the ability to infect and replicate in air-liquid interface cultures of human bronchial tracheal epithelial cells. These results suggest that circulating HCoVs in the field generally use cell-surface TMPRSS2 for cell entry, not endosomal cathepsins, in human airway epithelial cells. •Clinical isolates of HCoV-OC43 and -HKU1 were isolated from ALI-cultured HBTE cells.•Clinical isolates of HCoVs preferred the TMRRSS2 to cathepsins for cell entry.•Cell culture adapted HCoV-OC43 lost the ability to replicate in HBTE-ALI culture.