Abstract
Severe acute respiratory syndrome coronavirus (SARS-CoV) contains a single spike (S) protein, which binds to its receptor, angiotensin-converting enzyme 2 (ACE2), induces membrane fusion and serves as a neutralizing antigen. A SARS-CoV-S protein-bearing vesicular stomatitis virus (VSV) pseudotype using the VSVDeltaG* system was generated. Partial deletion of the SARS-CoV-S protein cytoplasmic domain allowed efficient incorporation into VSV particles and led to the generation of a pseudotype (VSV-SARS-St19) at high titre. Green fluorescent protein expression was demonstrated as early as 7 h after infection of Vero E6 cells with VSV-SARS-St19. VSV-SARS-St19 was neutralized by anti-SARS-CoV antibody and soluble ACE2, and its infection was blocked by treatment of Vero E6 cells with anti-ACE2 antibody. These results indicated that VSV-SARS-St19 infection is mediated by SARS-CoV-S protein in an ACE2-dependent manner. VSV-SARS-St19 will be useful for analysing the function of SARS-CoV-S protein and for developing rapid methods of detecting neutralizing antibodies specific for SARS-CoV infection.
MeSH Terms
Angiotensin-Converting Enzyme 2
Angiotensin-Converting Enzyme Inhibitors/pharmacology
Animals
Carboxypeptidases/antagonists & inhibitors,metabolism
Chlorocebus aethiops
Dose-Response Relationship, Drug
Hemagglutinins, Viral/metabolism,physiology
Humans
Membrane Glycoproteins/metabolism,physiology
Peptidyl-Dipeptidase A
Reassortant Viruses/metabolism,physiology
Rhabdoviridae Infections/virology
SARS Virus/chemistry,physiology
Spike Glycoprotein, Coronavirus
Vero Cells
Vesicular stomatitis Indiana virus/metabolism,physiology
Viral Envelope Proteins/metabolism,physiology
Virus Replication
Chemicals
Angiotensin-Converting Enzyme Inhibitors
Hemagglutinins, Viral
Membrane Glycoproteins
Spike Glycoprotein, Coronavirus
Viral Envelope Proteins
spike glycoprotein, SARS-CoV
spike protein, mouse hepatitis virus
Carboxypeptidases
Peptidyl-Dipeptidase A
ACE2 protein, human
Angiotensin-Converting Enzyme 2
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Fukushi Shuetsu
Special Pathogens Laboratory, Department of Virology I, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashimurayama, Tokyo 208-0011, Japan.
Mizutani Tetsuya
Special Pathogens Laboratory, Department of Virology I, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashimurayama, Tokyo 208-0011, Japan.
Saijo Masayuki
Special Pathogens Laboratory, Department of Virology I, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashimurayama, Tokyo 208-0011, Japan.
Matsuyama Shutoku
Laboratory of Respiratory Viral Diseases and SARS, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashimurayama, Tokyo 208-0011, Japan.
Miyajima Naoko
Laboratory of Respiratory Viral Diseases and SARS, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashimurayama, Tokyo 208-0011, Japan.
Taguchi Fumihiro
Laboratory of Respiratory Viral Diseases and SARS, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashimurayama, Tokyo 208-0011, Japan.
Itamura Shigeyuki
Laboratory of Influenza Virus, Department of Virology III, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashimurayama, Tokyo 208-0011, Japan.
Kurane Ichiro
Special Pathogens Laboratory, Department of Virology I, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashimurayama, Tokyo 208-0011, Japan.
Morikawa Shigeru
Special Pathogens Laboratory, Department of Virology I, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashimurayama, Tokyo 208-0011, Japan.
