Rapid seroconversion and persistent functional IgG antibodies in severe COVID-19 patients correlates with an IL-12p70 and IL-33 signature

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Abstract

Despite ongoing efforts to characterize the host response toward SARS-CoV-2, a major gap in our knowledge still exists regarding the magnitude and duration of the humoral response. Analysis of the antibody response in mild versus moderate/severe patients, using our new developed quantitative electrochemiluminescent assay for detecting IgM/IgA/IgG antibodies toward SARS-CoV-2 antigens, revealed a rapid onset of IgG/IgA antibodies, specifically in moderate/severe patients. IgM antibodies against the viral receptor binding domain, but not against nucleocapsid protein, were detected at early stages of the disease. Furthermore, we observed a marked reduction in IgM/IgA antibodies over-time. Adapting our assay for ACE2 binding-competition, demonstrated that the presence of potentially neutralizing antibodies is corelated with IgG/IgA. Finally, analysis of the cytokine profile in COVID-19 patients revealed unique correlation of an IL-12p70/IL33 and IgG seroconversion, which correlated with disease severity. In summary, our comprehensive analysis has major implications on the understanding and monitoring of SARS-CoV-2 infections.

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  1. SciScore for 10.1101/2020.06.28.20141838: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIRB: All experiments were reviewed and approved by the Helsinki committee (IRB#RMC-0265-20) and were performed according to their regulations and guidelines.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.

    Table 2: Resources

    Software and Algorithms
    SentencesResources
    Statistical analysis: All of the statistical analyses were performed using GraphPad Prism 8 software.
    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: We detected the following sentences addressing limitations in the study:
    This enzymatic activity introduces several limitations. Since the enzymatic reaction is time dependent, many ELISA tests require the use of a standard curve in order to detect the sample in the linear range of the assay. In electrochemiluminescence-based assays, such as the one we have developed, each individual sample in the plate is electronically excited and emits light, which is recorded immediately. This enables the assay to have a wide dynamic range that exceeds that of the standard ELISA tests. An additional advantage of the tests we developed is the high positive-to-negative ratio. Using a standard ELISA test, we could achieve a ∼10-fold induction, whereas using our assay we reached ∼100-fold. An additional advantage of the platform that we used is the ability to assess all three major antibody classes using multiplexing. This is extremely important in two different aspects. Regarding the diagnostic aspect, this allows us to increase the sensitivity of the assay by cross-analyzing the formation of different antibodies in each individual. Indeed, although many patients developed all three antibodies toward the viral RBD, several patients could generate only a single antibody class (e.g., they were positive for IgM but not for IgA or IgG). Such patients would be perceived as patients that did not develop antibodies if they were assessed by assays that enable the detection of 1 or even 2 antibodies. Strengthening this notion, our combined analysis strategy could increase...

    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.