Seasonal human coronavirus antibodies are boosted upon SARS-CoV-2 infection but not associated with protection

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Abstract

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  1. SciScore for 10.1101/2020.11.06.20227215: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    After 2 hours of incubation, ELISA plates were washed 3 times with PBS-T and bound antibodies were detected using a 1:5000 dilution of goat anti-human IgG conjugated to horseradish peroxidase (Jackson ImmunoResearch Laboratories, West Grove PA: cat. 109-036-098).
    anti-human IgG
    suggested: None
    The next day, heat inactivated serum samples were serially diluted 2-fold and mixed with 50-200 focus forming units/well of VSVΔG-RFP SARS-CoV-2 pseudotype virus and 600ng/ml of 1E9F9, a mouse anti-VSV Indiana G (Absolute Antibody, Oxford, UK: cat# Ab01402-2.0).
    anti-VSV
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Vero E6 cells stably expressing TMPRSS2 were seeded in 100μl at 2.5×104 cells/well in a 96 well collagen coated plate.
    Vero E6
    suggested: None
    The serum-virus mixture was incubated for 1 hour at 37⁰C before being plated on VeroE6 TMPRSS2 cells.
    VeroE6 TMPRSS2
    suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
    Software and Algorithms
    SentencesResources
    ELISA plates (Thermo Fisher Scientific: cat. 14-245-153) were coated overnight at 4°C with either 2 μg/mL SARS-CoV-2 antigen, 1.5 μg/mL hCOV antigen, or DPBS to control for background.
    Thermo Fisher Scientific
    suggested: (Thermo Fisher Scientific, RRID:SCR_008452)

    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. SciScore for 10.1101/2020.11.06.20227215: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementThe Penn Medicine BioBank is an established repository that routinely collects blood products from donors visiting the University of Pennsylvania Healthcare system upon written informed consent.RandomizationIn instances we did not find matched controls, we randomly selected patients with RT-PCR negative test results.Blindingnot detected.Power Analysisnot detected.Sex as a biological variablenot detected.Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    RESULTS Identification of SARS-CoV-2-reactive Antibodies in Human Sera Collected Prior to the COVID-19 Pandemic We completed ELISAs to quantify levels of pre-pandemic SARS-CoV-2-reactive IgG antibodies in 204 human serum samples collected in 2017.
    SARS-CoV-2-reactive IgG
    suggested: None
    Humans with Pre-pandemic SARS-CoV-2-reactive Antibodies Had Elevated Levels of Antibodies Against Previously Circulating Betacoronaviruses We completed ELISAs to quantify levels of pre-pandemic hCoV-reactive IgG antibodies in all 204 human serum samples collected in 2017.
    hCoV-reactive IgG
    suggested: None
    STAR METHODS KEY RESOURCES TABLE REAGENT or RESOURCE Antibodies Goat anti-human IgG-HRP mAb CR3022 mAb 1E9F9 Bacterial and Virus Strains SARS-CoV-2
    anti-human IgG-HRP
    suggested: None
    After 2 hours of incubation, ELISA plates were washed 3 times with PBS-T and bound antibodies were detected using a 1:5000 dilution of goat anti-human IgG conjugated to horseradish peroxidase (Jackson ImmunoResearch Laboratories, West Grove PA: cat. 109-036-098).
    anti-human IgG
    suggested: (Jackson ImmunoResearch Labs Cat# 109-036-098, RRID:AB_2337596)
    The next day, heat inactivated serum samples were serially diluted 2-fold and mixed with 50-200 focus forming units/well of VSVΔG-RFP SARS-CoV-2 pseudotype virus and 600ng/ml of 1E9F9, a mouse anti-VSV Indiana G (Absolute Antibody, Oxford, UK: cat# Ab01402-2.0).
    anti-VSV
    suggested: None
    We also compared antibody titers in pre-pandemic serum samples among SARS-CoV-2 PCR-confirmed individuals in relationship to hospitalization or need for respiratory support due to COVID-19.
    COVID-19
    suggested: None
    F-H) ELISAs were completed to quantify levels of serum antibodies binding to the full length S proteins from 229E, NL63, and OC43 using pre-pandemic serum samples with (n=12) and without (n=51).
    NL63
    suggested: None
    * D7 NL63 D0 D7 D0 D7 OC43 SARS-CoV-2 O SA C R SC oV -2 D0 Spike Fold-Change OC43 titer (day 7/day 0) * N L6 D7 229E * D0 C **** 9E 8.192 4.096 2.048 1.024 5.12 2.56 1.28 6.4 3.2 1.6 B **** Fold-Change titer (day 7/day 0) IgG antibody titer (reciprocal dilution) A Survived Died Figure 3.
    IgG
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    CRL-3216, RRID:CVCL_0063 Thermo Fisher cat.
    CRL-3216
    detected: (CCLV Cat# CCLV-RIE 1018, RRID:CVCL_0063)
    Cell lines 293F cells were from Thermo fisher (Thermo Fisher cat.
    293F
    suggested: RRID:CVCL_D615)
    . 293T cells were from ATCC (ATCC cat.
    293T
    suggested: KCB Cat# KCB 200744YJ, RRID:CVCL_0063)
    VeroE6/TMPRSS2 cells were a gift from Stefan Pohlman (German Primate Center, Leibniz Institute for Primate Research) as described previously17.
    VeroE6/TMPRSS2
    suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
    Vero E6 cells stably expressing TMPRSS2 were seeded in 100μl at 2.5x104 cells/well in a 96 well collagen coated plate.
    Vero E6
    suggested: None
    The serum-virus mixture was incubated for 1 hour at 37⁰C before being plated on VeroE6 TMPRSS2 cells.
    VeroE6 TMPRSS2
    suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
    Software and Algorithms
    SentencesResources
    STAR METHODS KEY RESOURCES TABLE REAGENT or RESOURCE Antibodies Goat anti-human IgG-HRP mAb CR3022 mAb 1E9F9 Bacterial and Virus Strains SARS-CoV-2
    STAR
    suggested: (STAR, RRID:SCR_015899)
    The Penn Medicine BioBank is an established repository that routinely collects blood products from donors visiting the University of Pennsylvania Healthcare system upon written informed consent.
    Pennsylvania Healthcare
    suggested: None
    ELISA plates (Thermo Fisher Scientific: cat. 14- 245-153) were coated overnight at 4oC with either 2 μg/mL SARS-CoV-2 antigen, 1.5 μg/mL hCOV antigen, or DPBS to control for background.
    Thermo Fisher Scientific
    suggested: (Thermo Fisher Scientific, RRID:SCR_008452)
    QUANTIFICATION AND STATISTICAL ANALYSIS Statistical analyses were performed using Prism version 8 (GraphPad Software, San Diego CA)
    Prism
    suggested: (PRISM, RRID:SCR_005375)
    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 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.