High titers of multiple antibody isotypes against the SARS-CoV-2 spike receptor-binding domain and nucleoprotein associate with better neutralization

  1. Maria G. Noval
  2. Maria E. Kaczmarek
  3. Akiko Koide
  4. Bruno A. Rodriguez-Rodriguez
  5. Ping Louie
  6. Takuya Tada
  7. Takamitsu Hattori
  8. Tatyana Panchenko
  9. Larizbeth A. Romero
  10. Kai Wen Teng
  11. Andrew Bazley
  12. Maren de Vries
  13. Marie I. Samanovic
  14. Jeffrey N. Weiser
  15. Ioannis Aifantis
  16. Joan Cangiarella
  17. Mark J. Mulligan
  18. Ludovic Desvignes
  19. Meike Dittmann
  20. Nathaniel R. Landau
  21. Maria Aguero-Rosenfeld
  22. Shohei Koide
  23. Kenneth A. Stapleford

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Abstract

Understanding antibody responses to SARS-CoV-2 is indispensable for the development of containment measures to overcome the current COVID-19 pandemic. Here, we determine the ability of sera from 101 recovered healthcare workers to neutralize both authentic SARS-CoV-2 and SARS-CoV-2 pseudotyped virus and address their antibody titers against SARS-CoV-2 nucleoprotein and spike receptor-binding domain. Interestingly, the majority of individuals have low neutralization capacity and only 6% of the healthcare workers showed high neutralizing titers against both authentic SARS-CoV-2 virus and the pseudotyped virus. We found the antibody response to SARS-CoV-2 infection generates antigen-specific isotypes as well as a diverse combination of antibody isotypes, with high titers of IgG, IgM and IgA against both antigens correlating with neutralization capacity. Importantly, we found that neutralization correlated with antibody titers as quantified by ELISA. This suggests that an ELISA assay can be used to determine seroneutralization potential. Altogether, our work provides a snapshot of the SARS-CoV-2 neutralizing antibody response in recovered healthcare workers and provides evidence that possessing multiple antibody isotypes may play an important role in SARS-CoV-2 neutralization.

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  1. ScreenIT

    SciScore for 10.1101/2020.08.15.252353: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementConsent: All patients gave written consent and all samples were deidentified for this study under IRB #i20-00595 (SARS-CoV-2 infected) and IRB #s18-02037 (healthy controls).
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line AuthenticationContamination: All cells were maintained at 37°C with 5% CO2 and confirmed mycoplasma free.

    Table 2: Resources

    Antibodies
    SentencesResources
    Serum IgG, IgA, and IgM antibodies to the SARS-CoV-2 nucleoprotein were tested using the SARS-CoV-2 IgG, IgA and IgM ELISA kits manufactured by Virotech Diagnostics GmbH for Gold Standard Diagnostics (Davis, CA) following the manufacturer’s instructions.
    IgM
    suggested: None
    SARS-CoV-2 IgG
    suggested: None
    15 µl of secondary antibody (anti human Fab-HRP: Jackson ImmunoResearch, cat#
    anti human
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Human embryonic kidney (HEK) 293T cells were maintained in DMEM supplemented with 10% FBS, and 1% penicillin/streptomycin (P/S).
    HEK
    suggested: None
    Expi293T cells (Thermo Fisher) were maintained in Expi293 Expression Medium (Thermo Fisher).
    Expi293T
    suggested: None
    Viral titers were determined by plaque assay in Vero E6 cells.
    Vero E6
    suggested: RRID:CVCL_XD71)
    Pseudotyped virus preparation: Pseudotyped virus was produced by calcium phosphate cotransfection of 293T cells with pMDL, plenti.GFP-NLuc, pcSARS-CoV-2-SΔ19 and pcRev at a ratio of 4:3:4:1.
    293T
    suggested: None
    Pseudotype virus neutralization assay: To determine neutralizing serum titers, ACE2-293T cells were plated in 96 well tissue culture dishes at 10,000 cells/well.
    ACE2-293T
    suggested: None
    Software and Algorithms
    SentencesResources
    Data analysis and statistics: All experiments were performed in technical duplicates and data analysis and statistics were performed using GraphPad Prism (Version 8.4.3)
    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.
    • No funding statement was detected.
    • 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.

  2. Version published to 10.1101/2020.08.15.252353v1 on bioRxiv