Functional convalescent plasma antibodies and pre-infusion titers shape the early severe COVID-19 immune response

  1. Jonathan D. Herman
  2. Chuangqi Wang
  3. Carolin Loos
  4. Hyunah Yoon
  5. Johanna Rivera
  6. M. Eugenia Dieterle
  7. Denise Haslwanter
  8. Rohit K. Jangra
  9. Robert H. Bortz
  10. Katharine J. Bar
  11. Boris Julg
  12. Kartik Chandran
  13. Douglas Lauffenburger
  14. Liise-anne Pirofski
  15. Galit Alter

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Abstract

Transfer of convalescent plasma (CP) had been proposed early during the SARS-CoV-2 pandemic as an accessible therapy, yet trial results worldwide have been mixed, potentially due to the heterogeneous nature of CP. Here we perform deep profiling of SARS-CoV-2-specific antibody titer, Fc-receptor binding, and Fc-mediated functional assays in CP units, as well as in plasma from hospitalized COVID-19 patients before and after CP administration. The profiling results show that, although all recipients exhibit expanded SARS-CoV-2-specific humoral immune responses, CP units contain more functional antibodies than recipient plasma. Meanwhile, CP functional profiles influence the evolution of recipient humoral immunity in conjuncture with the recipient’s pre-existing SARS-CoV2-specific antibody titers: CP-derived SARS-CoV-2 nucleocapsid-specific antibody functions are associated with muted humoral immune evolution in patients with high titer anti-spike IgG. Our data thus provide insights into the unexpected impact of CP-derived functional anti-spike and anti-nucleocapsid antibodies on the evolution of SARS-CoV-2-specific response following severe infection.

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  1. Version published to 10.1038/s41467-021-27201-y
  2. ScreenIT

    SciScore for 10.1101/2021.03.08.21253157: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    RandomizationFor high S_IgG1 or low S_IgG1 group respectively, we broke the recipient donor pair, randomly matched them, and calculated the Spearman correlation on the permuted recipient-donor pair 1000 times.
    BlindingAll experiments were performed in duplicate while operators were blinded to study group assignment and all cases and controls were run at the same time to avoid batch effects.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Antibody Titer and Fc-Receptor Binding Assays: Antigen-specific antibody subclass, isotype, and Fc-receptor (FcR) binding levels were assayed with a customized multiplexed Luminex bead array, as previously described74.
    Antigen-specific antibody subclass, isotype,
    suggested: None
    Antigen-specific antibody titers were detected with Phycoerythrin (PE)-coupled antibodies against IgG1, IgG2, IgG3, IgG4, IgA1, and IgM (SouthernBiotech, Birmingham, AL).
    Phycoerythrin ( PE)-coupled antibodies against IgG1
    suggested: None
    IgG1
    suggested: None
    IgG2
    suggested: None
    IgG3
    suggested: None
    IgG4
    suggested: None
    IgA1
    suggested: None
    IgM ( SouthernBiotech , Birmingham , AL)
    suggested: None
    IgM
    suggested: None
    Fluorescence was detected using an Intellicyt iQue with a 384-well handling robot (PAA) and analyzed using Forecyt software by gating on fluorescent bead regions and PE mean fluorescence intensity (MFI) was measured as the readout of each antigen-specific antibody measurements.
    antigen-specific
    suggested: None
    Functional Assays: Bead-based assays were used to quantify antibody-dependent cellular phagocytosis (ADCP), antibody-dependent neutrophil phagocytosis (ADNP) and antibody-dependent complement deposition (ADCD) in the MGH SARS-CoV-2 cohort, as previously described75-79.
    antibody-dependent neutrophil phagocytosis ( ADNP
    suggested: None
    antibody-dependent complement deposition ( ADCD
    suggested: None
    Neutrophils were stained with an anti-CD66b PacBlue detection antibody (Biolegend) and fixed with 4% paraformaldehyde (Alfa Aesar).
    anti-CD66b
    suggested: None
    To measure antibody-dependent deposition of C3, lyophilized guinea pig complement (Cedarlane) was reconstituted according to manufacturer’s instructions and diluted in gelatin veronal buffer with calcium and magnesium (GBV++) (Boston BioProducts) and mixed with immune complexes.
    C3
    suggested: None
    After a 20-minute incubation at 37C, C3 was detected with an anti-C3 fluorescein-conjugated goat IgG fraction detection antibody (Mpbio).
    anti-C3 fluorescein-conjugated goat IgG
    suggested: None
    NK cells were mixed with a staining cocktail containing anti-CD107a BV605 antibody (Biolegand), Golgi stop (BD Biosciences) and Brefeldin A (BFA, Sigma Aldrich).
    anti-CD107a
    suggested: (BioLegend Cat# 328633, RRID:AB_2562649)
    Subsequently, cells were permeabilized using Perm B (Invitrogen) and intracellularly stained with an anti-MIP1b-BV421 (BD Biosciences) and IFNg-PE (BioLegend) antibodies.
    anti-MIP1b-BV421
    suggested: None
    IFNg-PE
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Briefly, CCP samples were serially diluted and incubated with pre-titrated amounts of virus for 1 hr at RT, plasma-virus mixtures were added to 96-wellplates (Corning) containing monolayers of Vero cells, incubated for 7hr at 37°C/5% CO2, fixed with 4% paraformaldehyde (Sigma) in PBS, washed with PBS, and stored in PBS containing Hoechst-33342 (1:2,000 dilution; Invitrogen)
    Vero
    suggested: None
    Software and Algorithms
    SentencesResources
    Viral infectivity was measured by automated enumeration of green fluorescent protein (GFP)-positive cells from captured images using a Cytation5 automated fluorescence microscope (BioTek) and analyzed using the Gen5 data analysis software (BioTek).
    Gen5
    suggested: (Gen5, RRID:SCR_017317)
    The serum half-maximal inhibitory concentration (IC50) was calculated using a nonlinear regression analysis with GraphPad Prism software.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Additionally, the labels and positions of nodes and links were adjusted for better visualization using the software Adobe Illustrator 2020 (24.2.3).
    Adobe Illustrator
    suggested: (Adobe Illustrator, RRID:SCR_010279)

    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.

  3. Version published to 10.1101/2021.03.08.21253157 on medRxiv