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Human airway cells prevent SARS-CoV-2 multibasic cleavage site cell culture adaptation.
Lamers, Mart M; Mykytyn, Anna Z; Breugem, Tim I; Wang, Yiquan; Wu, Douglas C; Riesebosch, Samra; van den Doel, Petra B; Schipper, Debby; Bestebroer, Theo; Wu, Nicholas C; Haagmans, Bart L.
  • Lamers MM; Viroscience Department, Erasmus Medical Center, Rotterdam, Netherlands.
  • Mykytyn AZ; Viroscience Department, Erasmus Medical Center, Rotterdam, Netherlands.
  • Breugem TI; Viroscience Department, Erasmus Medical Center, Rotterdam, Netherlands.
  • Wang Y; Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, United States.
  • Wu DC; Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, United States.
  • Riesebosch S; Viroscience Department, Erasmus Medical Center, Rotterdam, Netherlands.
  • van den Doel PB; Viroscience Department, Erasmus Medical Center, Rotterdam, Netherlands.
  • Schipper D; Viroscience Department, Erasmus Medical Center, Rotterdam, Netherlands.
  • Bestebroer T; Viroscience Department, Erasmus Medical Center, Rotterdam, Netherlands.
  • Wu NC; Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, United States.
  • Haagmans BL; Center for Biophysics and Quantitative Biology, University of Illinois at Urbana-Champaign, Urbana, United States.
Elife ; 102021 04 09.
Article in English | MEDLINE | ID: covidwho-1389777
Preprint
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ABSTRACT
Virus propagation methods generally use transformed cell lines to grow viruses from clinical specimens, which may force viruses to rapidly adapt to cell culture conditions, a process facilitated by high viral mutation rates. Upon propagation in VeroE6 cells, SARS-CoV-2 may mutate or delete the multibasic cleavage site (MBCS) in the spike protein. Previously, we showed that the MBCS facilitates serine protease-mediated entry into human airway cells (Mykytyn et al., 2021). Here, we report that propagating SARS-CoV-2 on the human airway cell line Calu-3 - that expresses serine proteases - prevents cell culture adaptations in the MBCS and directly adjacent to the MBCS (S686G). Similar results were obtained using a human airway organoid-based culture system for SARS-CoV-2 propagation. Thus, in-depth knowledge on the biology of a virus can be used to establish methods to prevent cell culture adaptation.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Virus Cultivation / Epithelial Cells / Virus Internalization / Spike Glycoprotein, Coronavirus / SARS-CoV-2 Limits: Animals / Humans Language: English Year: 2021 Document Type: Article Affiliation country: ELIFE.66815

Full text: Available Collection: International databases Database: MEDLINE Main subject: Virus Cultivation / Epithelial Cells / Virus Internalization / Spike Glycoprotein, Coronavirus / SARS-CoV-2 Limits: Animals / Humans Language: English Year: 2021 Document Type: Article Affiliation country: ELIFE.66815