Antibody Response against SARS-CoV-2 and Seasonal Coronaviruses in Nonhospitalized COVID-19 Patients

  1. Natalia Ruetalo
  2. Ramona Businger
  3. Karina Althaus
  4. Simon Fink
  5. Felix Ruoff
  6. Michaela Pogoda
  7. Angelika Iftner
  8. Tina Ganzenmüller
  9. Klaus Hamprecht
  10. Bertram Flehmig
  11. Tamam Bakchoul
  12. Markus F. Templin
  13. Michael Schindler

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Abstract

There is strong interest in the nature of the neutralizing antibody response against SARS-CoV-2 in infected individuals. For vaccine development, it is especially important which antibodies confer protection against SARS-CoV-2, if there is a phenomenon called antibody-dependent enhancement (ADE) of infection, and if there is cross-protection by antibodies directed against seasonal coronaviruses.

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  1. ScreenIT

    SciScore for 10.1101/2020.08.07.20169961: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementConsent: Study participants and sample processing: Blood was drawn from potential blood donors for reconvalescent plasma therapy after written consent at the Clinical Transfusion Medicine, Tübingen between April 04 and May 12, 2020, under the guidelines of the local ethics committees 222/2020BO.
    IRB: Study participants and sample processing: Blood was drawn from potential blood donors for reconvalescent plasma therapy after written consent at the Clinical Transfusion Medicine, Tübingen between April 04 and May 12, 2020, under the guidelines of the local ethics committees 222/2020BO.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Supernatant as well as cell lysates from infected cells were tested by Western blot using a SARS-CoV-2 anti-nucleocapsid protein (NP) specific antibody (GeneTex).
    anti-nucleocapsid protein (NP
    suggested: None
    Mediagnost, IgG antibody detection directed to the S1 RBD SARS-CoV-2 in human sera using Mediagnost test system was made according to the manufacturer’s instructions.
    IgG
    suggested: None
    Next a horseradish peroxidase (HRP) conjugated goat anti-human IgG binds to the human IgG antibodies.
    anti-human IgG
    suggested: None
    Increasing extinctions represent increasing amounts of antibodies to SARS-CoV-2 S1 protein.
    SARS-CoV-2 S1 protein.
    suggested: None
    IgA and IgM antibody detection directed to the SARS-CoV-2 S1 protein was made in analogy to the above-described IgG detection system except that the HRP labeled detection antibody was directed against human IgA or human IgM antibodies.
    IgA
    suggested: None
    IgM
    suggested: None
    human IgA
    suggested: None
    human IgM
    suggested: None
    Elecsys anti-SARS-CoV-2 (Roche): For qualitative detection of anti SARS-CoV-2 (IgG+IgM) antibodies the electrochemiluminescence immunoassay (ECLIA) was performed using the fully automated cobas e 6000/601 immunoassay analyzer (Roche Diagnostics, Mannheim,
    anti-SARS-CoV-2
    suggested: None
    anti SARS-CoV-2 (IgG+IgM
    suggested: None
    For positive control, we used a dilution series of a serum from a reconvalescent student infected symptomatically (fever, cough, loss of smell) tested posiive for viral RNA and NC-specific antibodies.
    NC-specific
    suggested: None
    Multiplexed detection of anti-coronavirus antibodies: Whole viral protein lysates from 229E, OC43, and NL63 (ZeptoMetrix Corp) and from SARS-CoV-2 were used for DigiWest as described (16).
    anti-coronavirus
    suggested: None
    NL63
    suggested: None
    Cells were incubated for 1 h at rt with 100 µl of serum from a hospitalized convalescent donor in a 1:1000 dilution and washed 3 times with PBS. 100 µl of goat anti-human Alexa594 1:2000 in PBS was used as secondary antibody for 1h at rt.
    anti-human
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Neutralization assay: For neutralization experiments, 1×104 Caco-2 cells/well were seeded in 96-well plates the day before infection in media containing 5% FCS.
    Caco-2
    suggested: None
    Software and Algorithms
    SentencesResources
    Virus neutralizing titers (VNT50) were calculated as the half-maximal inhibitory dose (ID50) using 4-parameter nonlinear regression (GraphPad Prism)
    GraphPad
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Software and statistical analysis: GraphPad Prism 8.0 was used for statistical and correlation analyses and to generate graphs.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Other software used included Gen5 v.3.04.
    Gen5
    suggested: (Gen5, RRID:SCR_017317)

    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.

  2. Version published to 10.1128/msphere.01145-20
  3. Version published to 10.1101/2020.08.07.20169961v2 on medRxiv
  4. Version published to 10.1101/2020.08.07.20169961v1 on medRxiv