Anti-SARS-CoV-2 IgG and IgA antibodies in COVID-19 convalescent plasma do not facilitate antibody-dependent enhance of viral infection

  1. Natasha M Clark
  2. Sanath Kumar Janaka
  3. William Hartman
  4. Susan Stramer
  5. Erin Goodhue
  6. John Weiss
  7. David T. Evans
  8. Joseph P. Connor

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Abstract

The novel coronavirus SARS-CoV2, which causes COVID-19, has resulted in the death of nearly 4 million people within the last 18 months. While preventive vaccination and monoclonal antibody therapies have been rapidly developed and deployed, early in the pandemic the use of COVID-19 convalescent plasma (CCP) was a common means of passive immunization, with the theoretical risk of antibody-dependent enhancement (ADE) of viral infection remaining undetermined. Though vaccines elicit a strong and protective immune response, and transfusion of CCP with high titers of neutralization activity are correlated with better clinical outcomes, the question of whether antibodies in CCP can enhance infection of SARS-CoV2 has not been directly addressed. In this study, we analyzed for and observed passive transfer of neutralization activity with CCP transfusion. Furthermore, to specifically understand if antibodies against the spike protein (S) enhance infection, we measured the anti-S IgG, IgA, and IgM responses and adapted retroviral-pseudotypes to measure virus neutralization with target cells expressing the ACE2 virus receptor and the Fc alpha receptor (FcαR) or Fc gamma receptor IIA (FcγRIIA). Whereas neutralizing activity of CCP correlated best with higher titers of anti-S IgG antibodies, the neutralizing titer was not affected when Fc receptors were present on target cells. These observations support the absence of antibody-dependent enhancement of infection (ADE) by IgG and IgA isotypes found in CCP. The results presented, therefore, support the clinical use of currently available antibody-based treatment including the continued study of CCP transfusion strategies.

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    SciScore for 10.1101/2021.09.14.460394: (What is this?)

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

    Table 1: Rigor

    EthicsIRB: CCP recipients and clinical parameters: This study was approved by the University of Wisconsin Institutional Review Board.
    Consent: Cases met all criteria for enrollment under the Mayo Clinic Expanded Access Protocol (IND # 20-003312) (EAP) and gave written, informed consent for CCP transfusion and data collection.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Rhesus Anti-SARS CoV Spike monoclonal antibody (NHP Reagent Resource)
    Anti-SARS
    suggested: None
    anti-Dengue monoclonal antibody, and 6 negative control plasma samples were added to each plate for validation.
    anti-Dengue
    suggested: None
    Area under the curve (AUC) was calculated (Graphpad Prism) as a measure of the anti-S antibody titers.
    anti-S
    suggested: None
    Anti-SARS-CoV2 Ig assay (CoV2T) (Ortho Clinical Diagnostics, Markham, Ontario): This assay is an automated assay that uses a two stage immunometric technique to identify anti-SARS-CoV2 Spike protein antibodies (IgG and IgM).
    anti-SARS-CoV2 Spike protein antibodies ( IgG
    suggested: None
    IgM
    suggested: None
    A cut-off threshold value was defined to determine a reactive sample and the signal to cut-off value (S/CO) was used as a quantitative measure of anti-spike antibodies.
    anti-spike
    suggested: None
    Architect Anti-SARS-CoV2 IgG assay (Abbott Laboratories, Chicago, IL): This assay is an automated, two-step immunoassay for the detection of IgG antibodies to SARS-CoV2 nucleocapsid proteins.
    Anti-SARS-CoV2 IgG
    suggested: None
    SARS-CoV2 nucleocapsid proteins .
    suggested: None
    Antibodies are captured on antigen coated paramagnetic microparticles and then detected by incubation with an acridinium-conjugated anti-human IgG.
    anti-human IgG
    suggested: None
    Although neither of these assays are used clinically as a quantitative test for the purposes of this study both the S/CO and the Index were considered to be reflective of the amount of anti-SARS-CoV2 antibodies in each sample.
    anti-SARS-CoV2
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    . 293T cells were transduced with retroviral particles carrying a pQCXIP vector encoding the gene for the human ACE2 protein, then selected and maintained in D10 supplemented with 1µg/ml Puromycin.
    293T
    suggested: None
    The 293T-ACE2 cells were transduced with retroviral particles carrying pQCXIH vector encoding the gene for FcαR and selected in D10 supplemented with 100μg/ml Hygromycin B and 1μg/ml Puromycin.
    293T-ACE2
    suggested: None
    Expression of FcαR and ACE2 in 293T-ACE2-FcαR cells were confirmed by flow cytometry.
    293T-ACE2-FcαR
    suggested: None
    Recombinant DNA
    SentencesResources
    The 293T-ACE2 cells were transduced with retroviral particles carrying pQCXIH vector encoding the gene for FcαR and selected in D10 supplemented with 100μg/ml Hygromycin B and 1μg/ml Puromycin.
    pQCXIH
    suggested: RRID:Addgene_37104)
    Separately, the 293T-ACE2 cells were transduced with retroviral particles carrying pQCXIP vector encoding the gene for FcγRII.
    pQCXIP
    suggested: RRID:Addgene_15714)
    Software and Algorithms
    SentencesResources
    Area under the curve (AUC) was calculated (Graphpad Prism) as a measure of the anti-S antibody titers.
    Graphpad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Architect Anti-SARS-CoV2 IgG assay (Abbott Laboratories, Chicago, IL): This assay is an automated, two-step immunoassay for the detection of IgG antibodies to SARS-CoV2 nucleocapsid proteins.
    Abbott Laboratories
    suggested: None
    All statistical analysis was done using the GraphPad Prism software (GraphPad Software, Inc La Jolla, CA).
    GraphPad
    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 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: 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.

    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.

  2. Version published to 10.1101/2021.09.14.460394v1 on bioRxiv