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SARS-CoV-2 growth, furin-cleavage-site adaptation and neutralization using serum from acutely infected hospitalized COVID-19 patients
- Source :
- Journal of General Virology, The Journal of General Virology, bioRxiv, article-version (status) pre, article-version (number) 2
- Publication Year :
- 2020
- Publisher :
- Microbiology Society, 2020.
-
Abstract
- Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019 (COVID-19), emerged at the end of 2019 and by mid-June 2020 the virus had spread to at least 215 countries, caused more than 8 000 000 confirmed infections and over 450 000 deaths, and overwhelmed healthcare systems worldwide. Like severe acute respiratory syndrome coronavirus (SARS-CoV), which emerged in 2002 and caused a similar disease, SARS-CoV-2 is a betacoronavirus. Both viruses use human angiotensin-converting enzyme 2 (hACE2) as a receptor to enter cells. However, the SARS-CoV-2 spike (S) glycoprotein has a novel insertion that generates a putative furin cleavage signal and this has been postulated to expand the host range. Two low-passage (P) strains of SARS-CoV-2 (Wash1 : P4 and Munich : P1) were cultured twice in Vero E6 cells and characterized virologically. Sanger and MinION sequencing demonstrated significant deletions in the furin cleavage signal of Wash1 : P6 and minor variants in the Munich : P3 strain. Cleavage of the S glycoprotein in SARS-CoV-2-infected Vero E6 cell lysates was inefficient even when an intact furin cleavage signal was present. Indirect immunofluorescence demonstrated that the S glycoprotein reached the cell surface. Since the S protein is a major antigenic target for the development of neutralizing antibodies, we investigated the development of neutralizing antibody titres in serial serum samples obtained from COVID-19 human patients. These were comparable regardless of the presence of an intact or deleted furin cleavage signal. These studies illustrate the need to characterize virus stocks meticulously prior to performing either in vitro or in vivo pathogenesis studies.
- Subjects :
- 0301 basic medicine
clinical isolates
viruses
030106 microbiology
adaptation
Virus Replication
Cleavage (embryo)
Article
Virus
03 medical and health sciences
Antigen
Neutralization Tests
Virology
Chlorocebus aethiops
Animals
Humans
Neutralizing antibody
Vero Cells
Furin
chemistry.chemical_classification
biology
Sequence Analysis, RNA
Animal
SARS-CoV-2
Genetic Variation
COVID-19
Viral Load
neutralization
Adaptation, Physiological
Antibodies, Neutralizing
In vitro
Immunoglobulin A
Hospitalization
030104 developmental biology
chemistry
Immunoglobulin G
Host-Pathogen Interactions
Proteolysis
Vero cell
biology.protein
RNA, Viral
Positive-strand RNA Viruses
furin cleavage
Antibody
Glycoprotein
Research Article
Subjects
Details
- Language :
- English
- ISSN :
- 14652099 and 00221317
- Database :
- OpenAIRE
- Journal :
- Journal of General Virology
- Accession number :
- edsair.doi.dedup.....7e4fdd90dd239f83ebea95bb14bb0c1d
- Full Text :
- https://doi.org/10.1099/jgv.0.001481