1887

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Volume 101, Issue 11

SARS-CoV-2 growth, furin-cleavage-site adaptation and neutralization using serum from acutely infected hospitalized COVID-19 patients Open Access

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 or pathogenesis studies.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): adaptation , clinical isolates , COVID-19 , furin cleavage , neutralization and SARS-CoV-2
Funding
This study was supported by the:
  • National Institutes of Health (Award AI060525)
    • Principle Award Recipient: Cynthia M McMillen
  • DSF Charitable Foundation
    • Principle Award Recipient: William B Klimstra
  • University of Pittsburgh Clinical and Translational Science Institute
    • Principle Award Recipient: William B Klimstra
  • Center for Vaccine Research, University of Pittsburgh
    • Principle Award Recipient: William B Klimstra
  • Dr. Patrick Gallagher, Chancellor of the University of Pittsburgh
    • Principle Award Recipient: William B Klimstra
© 2020 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. The Microbiology Society waived the open access fees for this article.
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2020-08-21
2024-04-25
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SARS-CoV-2 growth, furin-cleavage-site adaptation and neutralization using serum from acutely infected hospitalized COVID-19 patients
J. Gen. Virol. 101, 1156 (2020); https://doi.org/10.1099/jgv.0.001481
/content/journal/jgv/10.1099/jgv.0.001481
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