Human airway cells prevent SARS-CoV-2 multibasic cleavage site cell culture adaptation
- Erasmus MC, Netherlands
- University of Illinois at Urbana-Champaign, United States
- Invitae Corporation, United States
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.
Data availability
All scripts used for data processing are deposited in GitHub: https://github.com/wchnicholas/SARS_CoV2_mutation. Raw sequencing data are available at the NIH Short Read Archive under accession number: BioProject PRJNA694097.
Article and author information
Author details
Funding
Netherlands Organization for Health Research and Development (10150062010008)
- Bart L Haagmans
PPP allowance (LSHM19136)
- Bart L Haagmans
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2021, Lamers et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Human airway cells prevent SARS-CoV-2 multibasic cleavage site cell culture adaptation
eLife 10:e66815.
https://doi.org/10.7554/eLife.66815
Further reading
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- Microbiology and Infectious Disease
When culturing SARS-CoV-2 in the laboratory it is vital to avoid deletions in the gene for the spike protein that could affect the interpretation of experiments.
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- Epidemiology and Global Health
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- Microbiology and Infectious Disease
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