Switched and unswitched memory B cells detected during SARS-CoV-2 convalescence correlate with limited symptom duration

  1. Krista L. Newell
  2. Deanna C. Clemmer
  3. Justin B. Cox
  4. Yetunde I. Kayode
  5. Victoria Zoccoli-Rodriguez
  6. Harry E. Taylor
  7. Timothy P. Endy
  8. Joel R. Wilmore
  9. Gary M. Winslow

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Abstract

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), the causative agent of the pandemic human respiratory illness COVID-19, is a global health emergency. While severe acute disease has been linked to an expansion of antibody-secreting plasmablasts, we sought to identify B cell responses that correlated with positive clinical outcomes in convalescent patients. We characterized the peripheral blood B cell immunophenotype and plasma antibody responses in 40 recovered non-hospitalized COVID-19 subjects that were enrolled as donors in a convalescent plasma treatment study. We observed a significant negative correlation between the frequency of peripheral blood memory B cells and the duration of symptoms for convalescent subjects. Memory B cell subsets in convalescent subjects were composed of classical CD24 + class-switched memory B cells, but also activated CD24-negative and natural unswitched CD27 + IgD + IgM + subsets. Memory B cell frequency was significantly correlated with both IgG1 and IgM responses to the SARS-CoV-2 spike protein receptor binding domain (RBD) in most seropositive subjects. IgM + memory, but not switched memory, directly correlated with virus-specific antibody responses, and remained stable over 3 months. Our findings suggest that the frequency of memory B cells is a critical indicator of disease resolution, and that IgM + memory B cells may play an important role in SARS-CoV-2 immunity.

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  1. ScreenIT

    SciScore for 10.1101/2020.09.04.20187724: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementConsent: Exclusion criteria included the inability to give informed consent and/or an inability to donate plasma or blood transfusion in the past.
    IRB: Study approval: All participants provided written informed consent prior to participation in the study, which was performed according to a protocol approved by the Institutional Review Board (IRB) of the SUNY Upstate Medical University under IRB number 1587400.
    Randomizationnot detected.
    BlindingAll samples were de-identified following collection, and researchers conducting assays were blinded to clinical data until final comparative analysis.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Flow Cytometry: The following antibodies used for flow cytometry were obtained from BioLegend: CD21 (Bu32), T-bet (4B10), CD38 (HIT2), CD11c (Bu15), CD3 (HIT3A), CD14 (HCD14), IgD (IA6-2), CD24 (MC5), IgM (MHM-88), CD27 (O323), and CD19 (HIB19).
    CD38
    suggested: (Agilent Cat# TC67401, RRID:AB_579635)
    CD11c
    suggested: (Agilent Cat# TC66501, RRID:AB_579626)
    CD3
    suggested: None
    CD14
    suggested: None
    IA6-2
    suggested: None
    CD24
    suggested: None
    CD27
    suggested: None
    CD19
    suggested: None
    HIB19
    suggested: None
    After incubation, plates were washed three times with PBS-T and HRP-conjugated secondary anti-human antibodies were used for detection.
    anti-human
    suggested: None
    IgM was detected using goat-anti-human-IgM-HRP (Sigma, #A6907), and IgG subclass antibodies were detected with mouse horseradish peroxidase (HRP)-conjugated anti-human IgG1 (9054-05), IgG2 (9060-05), IgG3 (9210-05), and IgG4 (9200-05) from SouthernBiotech.
    IgG subclass
    suggested: None
    anti-human IgG1
    suggested: (SouthernBiotech Cat# 9054-05, RRID:AB_2796627)
    anti-human IgG1 (9054-05)
    suggested: (SouthernBiotech Cat# 9054-05, RRID:AB_2796627)
    IgG2 (9060-05)
    suggested: (SouthernBiotech Cat# 9060-05, RRID:AB_2796633)
    IgG3 (9210-05)
    suggested: (SouthernBiotech Cat# 9210-05, RRID:AB_2796699)
    9210-05
    suggested: (SouthernBiotech Cat# 9210-05, RRID:AB_2796699)
    IgG4 (9200-05) from SouthernBiotech.
    suggested: None
    IgG4
    suggested: (SouthernBiotech Cat# 9200-05, RRID:AB_2796691)
    Experimental Models: Cell Lines
    SentencesResources
    Recombinant Twin-Strep-tagged RBD protein was purified from 293T cells transfected with paH-RBD SD1-3CH25, generously provided by Jason S.
    293T
    suggested: None
    Software and Algorithms
    SentencesResources
    Flow Cytometry: The following antibodies used for flow cytometry were obtained from BioLegend: CD21 (Bu32), T-bet (4B10), CD38 (HIT2), CD11c (Bu15), CD3 (HIT3A), CD14 (HCD14), IgD (IA6-2), CD24 (MC5), IgM (MHM-88), CD27 (O323), and CD19 (HIB19).
    BioLegend
    suggested: (BioLegend, RRID:SCR_001134)
    Data from stained samples were acquired using a BD Fortessa flow cytometer equipped with DIVA software (BD Biosciences) and were analyzed using FlowJo software (Tree Star).
    FlowJo
    suggested: (FlowJo, RRID:SCR_008520)
    Statistical Analysis: Statistical analyses were performed using GraphPad Prism software (v8.4.3).
    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: 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. Version published to 10.1371/journal.pone.0244855
  3. Version published to 10.1101/2020.09.04.20187724v2 on medRxiv
  4. Version published to 10.1101/2020.09.04.20187724v1 on medRxiv