Reduced neutralization of SARS-CoV-2 variants by convalescent plasma and hyperimmune intravenous immunoglobulins for treatment of COVID-19

  1. Juanjie Tang
  2. Youri Lee
  3. Supriya Ravichandran
  4. Gabrielle Grubbs
  5. Chang Huang
  6. Charles Stauft
  7. Tony Wang
  8. Basil Golding
  9. Hana Golding
  10. Surender Khurana

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Abstract

Hyperimmune immunoglobulin (hCoV-2IG) preparations generated from SARS-CoV-2 convalescent plasma (CP) are under evaluation in several clinical trials of hospitalized COVID-19 patients. Here we explored the antibody epitope repertoire, antibody binding and virus neutralizing capacity of six hCoV-2IG batches as well as nine convalescent plasma (CP) lots against SARS-CoV-2 and emerging variants of concern (VOC). The Gene-Fragment Phage display library spanning the SARS-CoV-2 spike demonstrated broad recognition of multiple antigenic sites spanning the entire spike including NTD, RBD, S1/S2 cleavage site, S2-fusion peptide and S2-heptad repeat regions. Antibody binding to the immunodominant epitopes was higher for hCoV-2IG than CP, with predominant binding to the fusion peptide. In the pseudovirus neutralization assay (PsVNA) and in the wild-type SARS-CoV-2 PRNT assay, hCoV-2IG lots showed higher titers against the WA-1 strain compared with CP. Neutralization of SARS-CoV-2 VOCs from around the globe were reduced to different levels by hCoV-2IG lots. The most significant loss of neutralizing activity was seen against the B.1.351 (9-fold) followed by P.1 (3.5-fold), with minimal loss of activity against the B.1.17 and B.1.429 (≤2-fold). Again, the CP showed more pronounced loss of cross-neutralization against the VOCs compared with hCoV-2IG. Significant reduction of hCoV-2IG binding was observed to the RBD-E484K followed by RBD-N501Y and minimal loss of binding to RBD-K417N compared with unmutated RBD. This study suggests that post-exposure treatment with hCoV-2IG is preferable to CP. In countries with co-circulating SARS-CoV-2 variants, identifying the infecting virus strain could inform optimal treatments, but would likely require administration of higher volumes or repeated infusions of hCOV-2IG or CP, in patients infected with the emerging SARS-CoV-2 variants.

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  1. ScreenIT

    SciScore for 10.1101/2021.03.19.436183: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIRB: This study was approved by the Food and Drug Administration’s Research Involving Human Subjects Committee (RIHSC #2020-04-02).
    Randomizationnot detected.
    BlindingThe GFPDL affinity selection was performed in duplicate (two independent experiments by research fellow in the lab, who was blinded to sample identity).
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Lentivirus pseudovirion neutralization assay (PsVNA): Antibody preparations were evaluated by SARS-CoV-2 pseudovirus neutralization assay (PsVNA) using WA-1 strain, UK variant (B.1.1.7 with spike mutations: H69-V70del, Y144del, N501Y, A570D, D614G, P681H, T716I, S982A, and D1118H), SA variant (B.1.351 strain with spike mutations L18F, D80A, D215G, L242-244del, R246I, K417N, E484K, N501Y, D614G, and A701V), CA variant (B.1.429 strain with spike mutations S13I, W152C, L452R, D614G) and JP variant (P.
    S982A
    suggested: None
    Affinity selection of SARS-CoV-2 GFPDL phages: Prior to panning of GFPDL with polyclonal hCoV-2IG antibodies, Ig components, which could non-specifically interact with phage proteins, were removed by incubation with UV-killed M13K07 phage-coated Petri dishes.
    hCoV-2IG
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    The PsVNA using 293-ACE2-TMPRSS2 cell line was described previously (10, 11).
    293-ACE2-TMPRSS2
    suggested: None
    Pseudovirions were produced by co-transfection Lenti-X 293T cells with psPAX2(gag/pol), pTrip-luc lentiviral vector and pcDNA 3.1 SARS-CoV-2-spike-deltaC19, using Lipofectamine 3000.
    293T
    suggested: None
    The supernatants were harvested at 48h post transfection and filtered through 0.45μm membranes and titrated using 293T-ACE2-TMPRSS2 cells (HEK293T cells that express ACE2 and TMPRSS2 proteins).
    293T-ACE2-TMPRSS2
    suggested: None
    HEK293T
    suggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)
    The virus-antibody mixture was then added to a 96-well plate with 5×104 Vero E6 cells.
    Vero E6
    suggested: RRID:CVCL_XD71)
    Software and Algorithms
    SentencesResources
    The maximum resonance units (Max RU) data shown in the figures were the calculated RU signal for the 1 mg/mL hCoV-2IG sample or 2019-IVIG or 10-fold dilution of CP. Statistical Analysis: All experimental data were analyzed in GraphPad Prism, version 9.0.1 (GraphPad software Inc, San Diego, CA) or R package.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Correlation analysis of PRNT and PsVNA titers were performed by computing Pearson’s correlation coefficient in 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 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 10.1101/2021.03.19.436183 on bioRxiv