Mouse-adapted SARS-CoV-2 protects animals from lethal SARS-CoV challenge

  1. Antonio Muruato
  2. Michelle N. Vu
  3. Bryan A. Johnson
  4. Meredith E. Davis-Gardner
  5. Abigail Vanderheiden
  6. Kumari Lokugamage
  7. Craig Schindewolf
  8. Patricia A. Crocquet-Valdes
  9. Rose M. Langsjoen
  10. Jessica A. Plante
  11. Kenneth S. Plante
  12. Scott C. Weaver
  13. Kari Debbink
  14. Andrew L. Routh
  15. David Walker
  16. Mehul S. Suthar
  17. Pei-Yong Shi
  18. Xuping Xie
  19. Vineet D. Menachery

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Abstract

The emergence of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has resulted in a pandemic causing significant damage to public health and the economy. Efforts to understand the mechanisms of Coronavirus Disease 2019 (COVID-19) have been hampered by the lack of robust mouse models. To overcome this barrier, we used a reverse genetic system to generate a mouse-adapted strain of SARS-CoV-2. Incorporating key mutations found in SARS-CoV-2 variants, this model recapitulates critical elements of human infection including viral replication in the lung, immune cell infiltration, and significant in vivo disease. Importantly, mouse adaptation of SARS-CoV-2 does not impair replication in human airway cells and maintains antigenicity similar to human SARS-CoV-2 strains. Coupled with the incorporation of mutations found in variants of concern, CMA3p20 offers several advantages over other mouse-adapted SARS-CoV-2 strains. Using this model, we demonstrate that SARS-CoV-2–infected mice are protected from lethal challenge with the original Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV), suggesting immunity from heterologous Coronavirus (CoV) strains. Together, the results highlight the use of this mouse model for further study of SARS-CoV-2 infection and disease.

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  1. Version published to 10.1371/journal.pbio.3001284
  2. ScreenIT

    SciScore for 10.1101/2021.05.03.442357: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    EthicsIACUC: Synthetic construction of SARS-CoV-2 mouse adapted strains were approved by the University of Texas Medical Branch Institutional Biosafety Committee.
    IRB: For samples Emory University, collection and processing were performed under approval from the University Institutional Review Board (IRB #00001080 and #00022371).
    Consent: Adults ≥18 years were enrolled who met eligibility criteria for SARS-CoV-2 infection (PCR or rapid antigen test confirmed by a commercially available assay) and provided informed consent.
    Sex as a biological variableAll patients enrolled in July 2020 and had a mean age of 57 (range: 26-85; 50% male).
    RandomizationNo blinding was used in any sample collections, nor were samples randomized.
    BlindingNo blinding was used in any sample collections, nor were samples randomized.
    Power Analysisnot detected.
    Cell Line AuthenticationAuthentication: Viral mutants were confirmed by sequence analysis prior to use.

    Table 2: Resources

    Antibodies
    SentencesResources
    IHC was conducted using rabbit polyclonal antibodies raised against the SARS-CoV nucleocapsid protein (clone NB100-56576 [1:100], Novus Biologicals, Littleton, CO) and biotinylated anti-rabbit IgG, streptavidin AP (Vector Labs, Burlingame, CA) with Permanent Red chromogen (Dako/Agilent, Santa Clara CA)
    SARS-CoV nucleocapsid protein
    suggested: (Creative Diagnostics Cat# DMAB8869, RRID:AB_2392503)
    The slides were incubated with anti-SARS-CoV antibody for one hour at room temperature, washed in 1X Tris-buffered saline (TBS), 0.05% Tween 20 (wash buffer), and incubated with goat biotinylated anti-rabbit IgG (H + L) antibody for 30 min at room temperature followed by rinse in wash buffer.
    anti-SARS-CoV
    suggested: None
    anti-rabbit IgG
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    The medium from electroporated cells as harvested at 40 hours post transfection and served as seed stocks for amplifying one passage on Vero E6 cells (P1 stock).
    Vero E6
    suggested: RRID:CVCL_XD71)
    Experimental Models: Organisms/Strains
    SentencesResources
    Mice and in vivo infection: Ten-week-old BALB/C mice were purchased from Charles River Laboratories and were maintained in Sealsafe™ HEPA-filtered air in/out units.
    BALB/C
    suggested: RRID:IMSR_ORNL:BALB/cRl)
    Software and Algorithms
    SentencesResources
    modeling: Spike receptor binding tables were constructed from a set of representative group 2B coronaviruses by using alignment data paired with neighbor-joining phylogenetic trees built in Geneious (v.9.1.5) using the spike amino acid sequences derived the following accession numbers: QHU79204 (SARS-CoV-2 WA1), AGZ48806 (RsSHC014), ALK02457 (WIV16), and AYV99817.1(SARS-CoV Urbani).
    Geneious
    suggested: (Geneious, RRID:SCR_010519)
    Homology models were visualized and manipulated in PyMOL (version 2.4.2).
    PyMOL
    suggested: (PyMOL, RRID:SCR_000305)
    Microsoft Excel for Mac 2011 was used to analyze data.
    Microsoft Excel
    suggested: (Microsoft Excel, RRID:SCR_016137)
    Pileup files were generated using Samtools v1.9 43 and minority variants were extracted by nucleotide voting (PHRED>=30) using a custom python3 script previously described 39.
    Samtools
    suggested: (SAMTOOLS, RRID:SCR_002105)

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

    Results from JetFighter: Please consider improving the rainbow (“jet”) colormap(s) used on page 22. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    Results from scite Reference Check: We found no unreliable references.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  3. Version published to 10.1101/2021.05.03.442357 on bioRxiv