Early antibody responses associated with survival in COVID19 patients

  1. Zhao-Hua Zhou
  2. Sai Dharmarajan
  3. Mari Lehtimaki
  4. Susan L. Kirshner
  5. Steven Kozlowski

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Abstract

Neutralizing antibodies to the SARS CoV-2 spike proteins have been issued Emergency Use Authorizations and are a likely mechanism of vaccines to prevent COVID-19. However, benefit of treatment with monoclonal antibodies has only been observed in clinical trials in outpatients with mild to moderate COVID-19 but not in patients who are hospitalized and/or have advanced disease. To address this observation, we evaluated the timing of anti SARS-CoV-2 antibody production in hospitalized patients with the use of a highly sensitive multiplexed bead-based immunoassay allowing for early detection of antibodies to SARS-CoV-2. We found significantly lower levels of antibodies to the SARS-CoV-2 spike protein in the first week after symptom onset in patients who expired as compared to patients who were discharged. We also developed a model to characterize the relationship between each patient’s individual antibody level trajectory and eventual COVID 19 outcome which can be adapted into a prediction model with more data.

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  1. Version published to 10.1371/journal.ppat.1009766
  2. ScreenIT

    SciScore for 10.1101/2021.02.21.21252168: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    BlindingThe patient manifestations, demographics and outcomes were blinded before evaluating sample antibody profiles detection and unblinded for correlation with outcomes after antibody levels were determined.
    Power Analysisnot detected.
    Sex as a biological variable, Figure S14 IgG Sensitivity Analysis (exclude 1000 and 1007) Anti-SARS CoV2 IgG Sensitivity Analysis Kinetics Figure S15 IgG Sensitivity Analysis MFI Week 1 Comparison Figure S16 IgG Sensitivity Analysis Modeling Table S5, Figure S17 IgG Sensitivity Analysis (Males) Anti-SARS CoV2 IgG Sensitivity Analysis Kinetics Figure S18 IgG Sensitivity Analysis MFI Week 1 Comparison Figure S19 IgG Sensitivity Analysis Modeling Table S6, Figure S20

    Table 2: Resources

    Antibodies
    SentencesResources
    , anti-human IgG-PE, anti-human IgM-V450, and biotinylated anti-human IgA monoclonal antibodies were purchased from BD Biosciences (San Jose, CA).
    anti-human IgG-PE
    suggested: (Santa Cruz Biotechnology Cat# sc-3756, RRID:AB_654591)
    biotinylated anti-human IgA monoclonal antibodies
    suggested: None
    anti-human IgA
    suggested: None
    Conjugation of antigens and antibody binding were confirmed by flow cytometry using ELISA-confirmed rabbit anti-SARS CoV2 immune serum, SARS CoV1 immune rabbit serum, non-immunized control rabbit serum, as well as convalescent human COVID19 serum and pre-COVID19 human serum samples.
    anti-SARS
    suggested: None
    In brief, serum samples were serially diluted with PBS buffer (containing 1% BSA and 2mM EDTA) from 1:10 up to 1:100,000,000, mixed with SARS-CoV-2 bead arrays, with and without free SARS-CoV-2 antigens, incubated at 4°C with shaking overnight; washed with PBS buffer, developed with anti-human Ig isotype antibodies, and detected by flow cytometry (BD LSRFortessa™).
    anti-human Ig isotype antibodies ,
    suggested: None
    The multiplexed beads array method was demonstrated to sensitively and specifically detect antibody signals in rabbit anti-SARS-CoV-2 immune serum, convalescent human COVID-19 serum, and serum samples from patients with PCR-confirmed COVID19.
    anti-SARS-CoV-2
    suggested: None
    Detection of anti-CoV2 antibody: Detection for anti-CoV2 antibodies in heat-inactivated serum or plasma samples was done using 96 well U-shaped plates for high-throughput runs.
    anti-CoV2
    suggested: None
    Due to limited titrations, antigen-specific antibody levels (i.e., extrapolated titer values) were calculated based on multiplying the inverse of each sample dilution by an additional inverse dilution factor and using the maximum value.
    antigen-specific
    suggested: None
    Table S1 IgG Analysis Using IgG Cut-point MFI Method Figure S6 IgG Cut-point MFI Week 1 Post-onset Comparison Figure S7 IgG Cut-point MFI Modeling Table S2, Figure S8 IgM Analysis Anti-SARS CoV2 IgM Antibody Kinetics Figure S9 IgM Week 1 Post-onset Comparison Figure S10 IgM Modeling Table S3,
    Anti-SARS CoV2 IgM
    suggested: None
    Figure S11 IgA Analysis Anti-SARS CoV2 IgA Antibody Kinetics Figure S12 IgA Week 1 Post-onset Comparison Figure S13 IgA Modeling Table S4,
    Anti-SARS CoV2 IgA
    suggested: None
    Software and Algorithms
    SentencesResources
    Flow data files were analyzed using FlowJo software (FlowJo, LLC, Ashland, OR).
    FlowJo
    suggested: (FlowJo, RRID:SCR_008520)

    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: We detected the following sentences addressing limitations in the study:
    This study has limitations, including the small number of patients and the evaluation of patients relatively early in the pandemic, with a high mortality. 10 out of 11 mortality outcomes were observed in males. Given our small sample size, we are not able to investigate if the relationship between antibody response and outcome was moderated by sex; However, there was a similar relationship when only male samples were evaluated (Supplementary Materials). Despite these limitations, a clear association between early antibody generation and survival is noted. These findings are consistent with other studies when matched for timeframe. These findings are robust across multiple Spike protein antigens and sensitivity analyses. This pattern was also observed with antibodies to RBD which are strongly correlated with neutralizing activity (23). A cut-point based maximum signal method had even stronger associations with early antibody levels (Supplementary Materials). Antibody data from past and future studies can be evaluated to further refine and confirm these findings; There may be other qualitative antibody characteristics associated with patient outcomes (24). Early antibody responses may relate to variability in time of infection to symptom onset and reflect other mechanisms related to disease onset. Although the mechanisms for these findings need further elucidation, these results align well with clinical studies on therapeutic antibodies (2). These antibodies show benefit in out...

    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 10.1101/2021.02.21.21252168 on medRxiv