No substantial preexisting B cell immunity against SARS-CoV-2 in healthy adults

  1. Meryem Seda Ercanoglu
  2. Lutz Gieselmann
  3. Sabrina Dähling
  4. Nareshkumar Poopalasingam
  5. Susanne Detmer
  6. Manuel Koch
  7. Michael Korenkov
  8. Sandro Halwe
  9. Michael Klüver
  10. Veronica Di Cristanziano
  11. Hanna Janicki
  12. Maike Schlotz
  13. Johanna Worczinski
  14. Birgit Gathof
  15. Henning Gruell
  16. Matthias Zehner
  17. Stephan Becker
  18. Kanika Vanshylla
  19. Christoph Kreer
  20. Florian Klein

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Abstract

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  1. Version published to 10.1016/j.isci.2022.103951
  2. ScreenIT

    SciScore for 10.1101/2021.09.08.459398: (What is this?)

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

    Table 1: Rigor

    EthicsIRB: Buffy coats from study participants were collected according to a study protocol approved by the Institutional Review Board of the University of Cologne (study protocol 16-054).
    Consent: All study participants provided informed consent.
    Sex as a biological variableThe study cohort comprised 150 individuals with 50.7% female and 49.3% male adults.
    RandomizationFor reverse transcription, sorted cells were incubated with 7 μl of a random-hexamer-primer master mix (RHP mix) consisting of 0.75 μl Random Hexamer Primer (Thermo Fisher Scientific), 0.5 μl NP-40 (Thermo Fisher Scientific), 0.15 μl RNaseOut (Thermo Fisher Scientific), and 5.6 μl of nuclease-free H2O at 65 °C for 1 min.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    1st PCR product was amplified using Q5 Hot Start High Fidelity DNA Polymerase (New England Biolabs) and specific forward and reverse primers including adaptor sequences which are homologous to the restriction sites of the antibody expression vector (IgG1, IgL, IgK (Tiller et al., 2008)).
    IgG1
    suggested: None
    IgL, IgK
    suggested: None
    The final concentration of purified antibodies was determined by UV/Vis spectroscopy using a Nanodrop (A280).
    A280
    suggested: None
    Anti-SARS-CoV-2 S1/S2 IgG and IgM antibody titers of plasma samples were also assessed using the automated DiaSorin’s LIAISON® SARS-CoV-2 S1/S2 protein ELISA kit according to the manufacturer’s instructions.
    Anti-SARS-CoV-2 S1/S2 IgG
    suggested: None
    IgM
    suggested: None
    Anti-SARS-CoV-2 S1 IgA antibody titers of plasma samples were also measured using the Euroimmun anti-SARS-CoV-2 ELISA on the automated system Euroimmun Analyzer I and S/CO values interpreted with following cut-off values: negative S/CO < 0.8, equivocal S/CO ≥ 0.8 - < 1.1, positive S/CO ≥ 1.1.
    Anti-SARS-CoV-2 S1 IgA
    suggested: None
    anti-SARS-CoV-2
    suggested: None
    Figure S4: Binding of monoclonal antibodies isolated from pre-pandemic blood samples, related to Figure 4 (A) Binding of monoclonal antibodies to SARS-CoV-2, HKU-1 and OC43 S proteins determined by serial dilution ELISA.
    HKU-1
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    HEK293-6E cells were maintained in FreeStyle 293 Expression Medium (Life Technologies) supplemented with 0.2 % penicillin/streptomycin under constant shaking at 90 – 120 rpm, 37 °C and 6 % CO2.
    HEK293-6E
    suggested: RRID:CVCL_HF20)
    VeroE6 and HEK293T-ACE2 cells were maintained in Dulbecco’s Modified Eagle Medium (Gibco) supplemented with 10 % fetal bovine serum (FBS, Sigma-Aldrich), 1 x penicillin-streptomycin (Gibco), 1 mM sodium pyruvate (Gibco) and 2 mM L-glutamine (Gibco) at 37 °C and 5 % CO2.
    HEK293T-ACE2
    suggested: None
    The sex of HEK293T, HEK293-6E and VeroE6 cell lines is female.
    VeroE6
    suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
    Briefly, expression constructs were transfected into the HEK293 EBNA cells using FuGENE HD transfection reagent (Promega).
    HEK293
    suggested: CLS Cat# 300192/p777_HEK293, RRID:CVCL_0045)
    Cell-surface-expressed S protein immunoassay: HEK293T cells were transfected with plasmids encoding the full-length SARS-CoV-2 S protein (GenBank ID.:
    HEK293T
    suggested: CCLV Cat# CCLV-RIE 1018, RRID:CVCL_0063)
    For testing neutralization at a single dilution, polyclonal IgG samples at a concentration of 1000 μg/ml, plasma samples at a dilution of 1:10, or mAbs at a concentration of 50 μg/ml, were co-incubated with pseudovirus supernatants for 1 h at 37°C, following which 293T-ACE-2 cells were added.
    293T-ACE-2
    suggested: RRID:CVCL_YZ65)
    Software and Algorithms
    SentencesResources
    Evaluations were performed using FlowJo10 software.
    FlowJo10
    suggested: None
    IC50 values were calculated as the antibody concentration causing a 50 % reduction in signal compared the virus-only controls using a dose-response curve in GraphPad Prism.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Statistical tests and analyses were done with GraphPad Prism (v7 and v8), Python (v3.6.8), R (v4.0.0) and Mircosoft Excel for Mac (v14.7.3 and 16.4.8).
    Python
    suggested: (IPython, RRID:SCR_001658)
    Mircosoft Excel
    suggested: None
    A two-tailed unpaired t test (Prism, GraphPad) was performed to test for the frequency differences of SARS-CoV-2-reactive IgG+ and IgG- B cells between pre-pandemic and convalescent blood samples (Figure 3A).
    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.

    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.

  3. Version published to 10.1101/2021.09.08.459398 on bioRxiv