Neutralizing Antibody and Soluble ACE2 Inhibition of a Replication-Competent VSV-SARS-CoV-2 and a Clinical Isolate of SARS-CoV-2

  1. James Brett Case
  2. Paul W. Rothlauf
  3. Rita E. Chen
  4. Zhuoming Liu
  5. Haiyan Zhao
  6. Arthur S. Kim
  7. Louis-Marie Bloyet
  8. Qiru Zeng
  9. Stephen Tahan
  10. Lindsay Droit
  11. Ma. Xenia G. Ilagan
  12. Michael A. Tartell
  13. Gaya Amarasinghe
  14. Jeffrey P. Henderson
  15. Shane Miersch
  16. Mart Ustav
  17. Sachdev Sidhu
  18. Herbert W. Virgin
  19. David Wang
  20. Siyuan Ding
  21. Davide Corti
  22. Elitza S. Theel
  23. Daved H. Fremont
  24. Michael S. Diamond
  25. Sean P.J. Whelan

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Abstract

No abstract available

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  1. Version published to 10.1016/j.chom.2020.06.021
  2. ScreenIT

    SciScore for 10.1101/2020.05.18.102038: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIRB: This study was approved by the Mayo Clinic Institutional Review
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Viral proteins were separated on a 8% acrylamide gel, transferred onto a nitrocellulose membrane, and incubated with human anti-SARS antibody CR3022 diluted in Tris-buffer saline containing 1% Tween-20 (TBS-T) and 5% milk, followed by incubation with HRP-conjugated goat anti-human antibody (Abcam) diluted in TBS-T containing 1% milk.
    anti-SARS
    suggested: None
    anti-human antibody
    suggested: None
    Samples were prescreened by the Euroimmun anti-SARS-CoV-2 IgG ELISA (Lubeck, Germany), a qualitative assay with the Food and Drug Administration Emergency Use Authorization that detects antibodies to the SARS-CoV-2 S protein.
    anti-SARS-CoV-2 IgG ELISA ( Lubeck , Germany)
    suggested: None
    Plates were washed and sequentially incubated with 1 μg/mL of CR3022 anti-S antibody (ter Meulen et al., 2006; Yuan et al., 2020) and HRP-conjugated goat anti-human IgG in PBS supplemented with 0.1% saponin and 0.1% BSA.
    anti-S
    suggested: None
    anti-human IgG
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    BSRT7/5, Vero CCL81, Vero E6, Vero E6-TMPRSS2, A549, Caco-2, Caco-2 BBe1
    A549
    suggested: NCI-DTP Cat# A549, RRID:CVCL_0023)
    Caco-2
    suggested: None
    , Huh7, HepG2, Hela, BHK-21, HEK293, and HEK293T were maintained in DMEM (Corning or VWR) supplemented with glucose, L-glutamine, sodium pyruvate, and 10% fetal bovine serum (FBS).
    Huh7
    suggested: None
    HepG2
    suggested: None
    BHK-21
    suggested: None
    HEK293
    suggested: None
    HT-29 cells were cultured in complete DMEM/F12 (Thermo-Fisher) supplemented with sodium pyruvate, non-essential amino acids, and HEPES.
    HT-29
    suggested: None
    Vero E6-TMPRSS2 cells were generated using a lentivirus vector.
    Vero E6-TMPRSS2
    suggested: None
    Briefly, HEK293T producer cells were transfected with pLX304-TMPRSS2, pCAGGS-VSV-G, and psPAX2, and cell culture supernatants were collected at 48 hours and clarified by centrifugation at 1,000 x g for 5 min.
    HEK293T
    suggested: None
    The resulting lentivirus was used to infect Vero E6 cells for 24 h, and cells were selected with 40 μg/ml blasticidin for 7 days.
    Vero E6
    suggested: None
    Briefly, BSRT7/5 cells were inoculated with vaccinia virus vTF7-3 and subsequently transfected with T7-expression plasmids encoding VSV N, P, L, and G, and an antigenomic copy of the viral genome.
    BSRT7/5
    suggested: CCLV Cat# CCLV-RIE 0583, RRID:CVCL_RW96)
    Next generation sequencing: Total RNA was extracted from Vero CCL81 cells infected with VSV-SARS-CoV-2-SΔ21 using Trizol (Invitrogen) according to the manufacturer’s protocol.
    Vero CCL81
    suggested: None
    Software and Algorithms
    SentencesResources
    Other plasmids were previously described: VSV N, P, L and G expression plasmids (Stanifer et al., 2011; Whelan et al., 1995), psPAX2 (Addgene), and pLX304-TMPRSS2 (Zang et al., 2020).
    Addgene
    suggested: (Addgene, RRID:SCR_002037)
    Alignment of each sample to VSV and SARS CoV-2 sequence was performed using bbmap v38.79
    bbmap
    suggested: (BBmap, RRID:SCR_016965)
    Mapped reads were extracted using samtools 1.9 and used for de novo assembly by SPAdes v3.13.0.
    samtools
    suggested: (SAMTOOLS, RRID:SCR_002105)
    SPAdes
    suggested: (SPAdes, RRID:SCR_000131)
    Consensus sequences for each RNA sample were generated by aligning contigs to the reference plasmid sequence pVSV(1+)-eGFP-SARS-CoV-2-S with SnapGene v5.0.
    SnapGene
    suggested: (SnapGene, RRID:SCR_015052)
    Data were processed using Prism software (GraphPad Prism 8.0).
    Prism
    suggested: (PRISM, RRID:SCR_005375)
    GraphPad
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Data were processed using Prism software (GraphPad Prism 8.0).
    Prism
    suggested: (PRISM, RRID:SCR_005375)
    GraphPad
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Statistical analyses: All statistical tests were performed using GraphPad Prism 8.0 software as described in the indicated figure legends.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)

    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 did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: We did not find any issues relating to colormaps.

    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.

    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 10.1101/2020.05.18.102038v1 on bioRxiv