SARS-CoV-2 pseudovirus infectivity and expression of viral entry-related factors ACE2, TMPRSS2, Kim-1, and NRP-1 in human cells from the respiratory, urinary, digestive, reproductive, and immune systems

J Med Virol. 2021 Dec;93(12):6671-6685. doi: 10.1002/jmv.27244. Epub 2021 Aug 4.

Abstract

Infection by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes a wide spectrum of syndromes involving multiple organ systems and is primarily mediated by viral spike (S) glycoprotein through the receptor-binding domain (RBD) and numerous cellular proteins including ACE2, transmembrane serine protease 2 (TMPRSS2), kidney injury molecule-1 (Kim-1), and neuropilin-1 (NRP-1). In this study, we examined the entry tropism of SARS-CoV-2 and SARS-CoV using S protein-based pseudoviruses to infect 22 cell lines and 3 types of primary cells isolated from respiratory, urinary, digestive, reproductive, and immune systems. At least one cell line or type of primary cell from each organ system was infected by both pseudoviruses. Infection by pseudoviruses is effectively blocked by S1, RBD, and ACE2 recombinant proteins, and more weakly by Kim-1 and NRP-1 recombinant proteins. Furthermore, cells with robust SARS-CoV-2 pseudovirus infection had strong expression of either ACE2 or Kim-1 and NRP-1 proteins. ACE2 glycosylation appeared to be critical for the infections of both viruses as there was a positive correlation between infectivity of either SARS-CoV-2 or SARS-CoV pseudovirus with the level of glycosylated ACE2 (gly-ACE2). These results reveal that SARS-CoV-2 cell entry could be mediated by either an ACE2-dependent or -independent mechanism, thus providing a likely molecular basis for its broad tropism for a wide variety of cell types.

Keywords: Angiotensin-converting enzyme 2 (ACE2); Kim-1; NRP-1; SARS-CoV; SARS-CoV-2; TMPRSS2; and glycosylation; pseudovirus; receptor binding domain (RBD); spike protein; viral entry tropism; virus receptor.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Angiotensin-Converting Enzyme 2 / metabolism*
  • Blotting, Western
  • COVID-19 / metabolism
  • COVID-19 / virology
  • Cell Line
  • Cells, Cultured
  • Fluorescent Antibody Technique
  • Gastrointestinal Tract / cytology
  • Gastrointestinal Tract / virology*
  • Genitalia / cytology
  • Genitalia / virology*
  • Hepatitis A Virus Cellular Receptor 1 / metabolism*
  • Humans
  • Immune System / cytology
  • Immune System / virology*
  • Neuropilin-1 / metabolism*
  • Respiratory System / cytology
  • Respiratory System / virology*
  • SARS-CoV-2 / physiology*
  • Serine Endopeptidases / metabolism*
  • Virus Internalization*

Substances

  • HAVCR1 protein, human
  • Hepatitis A Virus Cellular Receptor 1
  • NRP1 protein, human
  • Neuropilin-1
  • ACE2 protein, human
  • Angiotensin-Converting Enzyme 2
  • Serine Endopeptidases
  • TMPRSS2 protein, human