Functional SARS-CoV-2-Specific Immune Memory Persists after Mild COVID-19

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Abstract

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  1. SciScore for 10.1101/2020.08.11.20171843: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIRB: Ethics Statement: This study was approved by the University of Washington Institutional Review Board (Gale Lab, IRB 00009810).
    Consent: Informed consent was obtained from all enrolled participants.
    IACUC: All mouse experiments were performed in accordance with the University of Washington Institutional Care and Use Committee guidelines.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Controls (n=17) were selected based on having no prior positive SARS-CoV-2 PCR nasal swab and having no detectable SARS-CoV-2 RBD- or S-specific IgG or IgM plasma antibodies (within mean + 3 SD of 5 de-identified plasma samples drawn prior to 2020 generously donated by Wesley C.
    S-specific IgG
    suggested: None
    Secondary antibodies were diluted in dilution buffer as follows: anti-human IgG-HRP (Jackson ImmunoResearch) at 1:3000, anti-human IgM-HRP (Southern Biotech) at 1:3000, or anti-human IgA-HRP (Southern Biotech) at 1:1500.
    anti-human IgG-HRP
    suggested: None
    anti-human IgM-HRP
    suggested: (SouthernBiotech Cat# 2020-05, RRID:AB_2795603)
    anti-human IgA-HRP
    suggested: (SouthernBiotech Cat# 2050-05, RRID:AB_2687526)
    Experimental Models: Cell Lines
    SentencesResources
    After 30 minutes of incubation at 37°C, the plasma/virus mixtures were added to 12 well plates of Vero cells and incubated for 1 hour at 37C, rocking every 15 minutes.
    Vero
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    Tetramer validation in mice: Adult C57BL/6j mice (The Jackson Laboratory) were immunized with 50ug SARS-CoV-2 RBD in CFA in the footpad and, 7-10 days later, popliteal lymph nodes were dissected, mashed and stained in 10nM decoy-PE-APC tetramer and then 10nM RBD-PE as described17and as described below for immunophenotyping B cells.
    C57BL/6j
    suggested: None
    Software and Algorithms
    SentencesResources
    SARS-CoV-2 Protein Production and Purification: Tetramer generation: Recombinant trimeric spike and the RBD domain were both biotinylated using a EZ-Link Sulfo-NHS-LC Biotinylation Kit (ThermoFisher), tetramerized with streptavidin-PE (Agilent) and stored in 50% glycerol at −20°C as previously described17.
    ThermoFisher
    suggested: (ThermoFisher; SL 8; Centrifuge, RRID:SCR_020809)
    CR3022, a human SARS-CoV antibody previously determined to cross-react with SARS-CoV-2 was used as a positive control.
    SARS-CoV-2
    suggested: (Active Motif Cat# 91351, RRID:AB_2847848)
    Data was analysed in Prism (GraphPad).
    GraphPad
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Prism
    suggested: (PRISM, RRID:SCR_005375)
    Data was analysed in Prism (GraphPad).
    GraphPad
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Prism
    suggested: (PRISM, RRID:SCR_005375)

    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.

    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. SciScore for 10.1101/2020.08.11.20171843: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementMethods: Ethics Statement This study was approved by the University of Washington Institutional Review Board (Gale Lab, IRB 00009810).Randomizationnot detected.Blindingnot detected.Power Analysisnot detected.Sex as a biological variable) Sex 27% Male, 73Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Controls (n=17) were selected based on having no prior positive SARS-CoV-2 PCR nasal swab and having no detectable SARS-CoV-2 RBD- or S-specific IgG or IgM plasma antibodies (within mean + 3 SD of 5 de-identified plasma samples drawn prior to 2020 generously donated by Wesley C.
    S-specific IgG
    suggested: None
    Secondary antibodies were diluted in dilution buffer as follows: anti-human IgG-HRP (Jackson ImmunoResearch) at 1:3000, anti-human IgM-HRP (Southern Biotech) at 1:3000, or anti-human IgA-HRP (Southern Biotech) at 1:1500.
    anti-human IgG-HRP
    suggested: None
    anti-human IgM-HRP
    suggested: (SouthernBiotech Cat# 2020-05, RRID:AB_2795603)
    anti-human IgA-HRP
    suggested: (SouthernBiotech Cat# 2050-05, RRID:AB_2687526)
    Expression of recombinant full-length human IgG monoclonal antibodies were carried out in serum-free basal medium (Nutridoma-SP, Sigma-Aldrich).
    recombinant full-length human IgG monoclonal antibodies
    suggested: None
    human IgG
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    After 30 minutes of incubation at 37oC, the plasma/virus mixtures were added to 12 well plates of Vero cells and incubated for 1 hour at 37C, rocking every 15 minutes.
    Vero
    suggested: None
    For large scale transfections, expression vectors containing paired heavy and light chains were transiently transfected into 293T cells using polyethylenimine (PEI).
    293T
    suggested: KCB Cat# KCB 200744YJ, RRID:CVCL_0063)
    Experimental Models: Organisms/Strains
    SentencesResources
    Tetramer validation in mice Adult C57BL/6j mice (The Jackson Laboratory) were immunized with 50ug SARS-CoV-2 RBD in CFA in the footpad and, 7-10 days later, popliteal lymph nodes were dissected, mashed and stained in 10nM decoy-PE-APC tetramer and then 10nM RBD-PE as described17and as described below for immunophenotyping B cells.
    C57BL/6j
    suggested: None
    Software and Algorithms
    SentencesResources
    CR3022, a human SARS-CoV antibody previously determined to cross-react with SARS-CoV-2 was used as a positive control.
    SARS-CoV-2
    suggested: (Active Motif Cat# 91345, RRID:AB_2847847)
    Data was analysed in Prism (GraphPad).
    Prism
    suggested: (PRISM, RRID:SCR_005375)
    Stained samples were run on a LSRII flow cytometer and analyzed using FlowJo (Becton Dickinson).
    FlowJo
    suggested: (FlowJo, RRID:SCR_008520)
    Sequences were trimmed at Q30 using Geneious and submitted to IMGT/HighV-QUEST for alignment (Alamyar et al, 2012).
    Geneious
    suggested: (Geneious, RRID:SCR_010519)
    Statistics Statistics used are described in figure legends and were determined using Prism (Graphpad).
    Graphpad
    suggested: (GraphPad, RRID:SCR_000306)

    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 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.


    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.