Immunogenic profile of SARS-CoV-2 spike in individuals recovered from COVID-19

  1. Jennifer A Juno
  2. Hyon-Xhi Tan
  3. Wen Shi Lee
  4. Arnold Reynaldi
  5. Hannah G Kelly
  6. Kathleen Wragg
  7. Robyn Esterbauer
  8. Helen E Kent
  9. C Jane Batten
  10. Francesca L Mordant
  11. Nicholas A Gherardin
  12. Phillip Pymm
  13. Melanie H Dietrich
  14. Nichollas E Scott
  15. Wai-Hong Tham
  16. Dale I Godfrey
  17. Kanta Subbarao
  18. Miles P Davenport
  19. Stephen J Kent
  20. Adam K Wheatley

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Abstract

The rapid global spread of SARS-CoV-2 and resultant mortality and social disruption have highlighted the need to better understand coronavirus immunity to expedite vaccine development efforts. Multiple candidate vaccines, designed to elicit protective neutralising antibodies targeting the viral spike glycoprotein, are rapidly advancing to clinical trial. However, the immunogenic properties of the spike protein in humans are unresolved. To address this, we undertook an in-depth characterisation of humoral and cellular immunity against SARS-CoV-2 spike in humans following mild to moderate SARS-CoV-2 infection. We find serological antibody responses against spike are routinely elicited by infection and correlate with plasma neutralising activity and capacity to block ACE2/RBD interaction. Expanded populations of spike-specific memory B cells and circulating T follicular helper cells (cTFH) were detected within convalescent donors, while responses to the receptor binding domain (RBD) constitute a minor fraction. Using regression analysis, we find high plasma neutralisation activity was associated with increased spike-specific antibody, but notably also with the relative distribution of spike-specific cTFH subsets. Thus both qualitative and quantitative features of B and T cell immunity to spike constitute informative biomarkers of the protective potential of novel SARS-CoV-2 vaccines.

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  1. ScreenIT

    SciScore for 10.1101/2020.05.17.20104869: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIRB: Ethics Statement: The study protocols were approved by the University of Melbourne Human Research Ethics Committee (#2056689) and all associated procedures were carried out in accordance with the approved guidelines.
    Consent: All participants provided written informed consent in accordance with the Declaration of Helsinki.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Monoclonal antibodies for surface staining included: CD19-ECD (J3-119) (Beckman Coulter), CD20 Alexa700 (2H7), IgM-BUV395 (G20-127), CD21-BUV737 (B-ly4), IgD-Cy7PE (IA6-2), IgG-BV786 (G18-145) (BD), CD14-BV510 (M5E2), CD3-BV510 (OKT3), CD8a-BV510 (RPA-T8), CD16-BV510 (3G8), CD10-BV510 (HI10a), CD27-BV605 (O323) (Biolegend).
    CD21-BUV737
    suggested: None
    CD14-BV510
    suggested: None
    CD3-BV510
    suggested: None
    OKT3
    suggested: (BD Biosciences Cat# 566779, RRID:AB_2869862)
    CD8a-BV510
    suggested: None
    CD16-BV510
    suggested: None
    CD10-BV510
    suggested: None
    CD27-BV605
    suggested: None
    Following stimulation, cells were washed, stained with Live/dead Blue viability dye (ThermoFisher), and a cocktail of monoclonal antibodies: CD27 BUV737 (L128), CCR7 Alexa700 (150503), CD45RA PeCy7 (HI100), CD20 BUV805 (2H7), CD14 BUV395 (MOP9) (BD Biosciences), CD3 BV510 (SK7), CD4 BV605 (RPA-T4), CD8 BV650 (RPA-T8)
    CD27
    suggested: (BD Biosciences Cat# 742012, RRID:AB_2871310)
    CCR7
    suggested: (BD Biosciences Cat# 741981, RRID:AB_2871284)
    CD45RA
    suggested: (BD Biosciences Cat# 742052, RRID:AB_2871341)
    CD20
    suggested: (BD Biosciences Cat# 563781, RRID:AB_2744325)
    CD14
    suggested: (BD Biosciences Cat# 740241, RRID:AB_2739988)
    CD3
    suggested: None
    CD4
    suggested: None
    CD8
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    ) or HCoV-HKU1 S protein (isolate N5;residues 1 – 1290) were synthesised with furin cleavage site removed and P986/987 stabilisation mutations39, a C-terminal T4 trimerisation domain, Avitag and His-tag, expressed in Expi293 cells and purified by Ni-NTA affinity and size-exclusion chromatography using a Superose 6 16/70 column (GE Healthcare) (
    Expi293
    suggested: RRID:CVCL_D615)
    Residual virus infectivity in the plasma/virus mixtures was assessed in quadruplicate wells of Vero cells incubated in serum-free media containing 1 μg/ml TPCK trypsin at 37°C/5% CO2; viral cytopathic effect was read on day 5.
    Vero
    suggested: None
    Software and Algorithms
    SentencesResources
    Data are available via ProteomeXchange with identifier PXD019163.
    ProteomeXchange
    suggested: (ProteomeXchange, RRID:SCR_004055)
    Pairwise correlations were assessed using Spearmans tests in Prism 8.0 (Graphpad).
    Prism
    suggested: (PRISM, RRID:SCR_005375)
    Graphpad
    suggested: (GraphPad, RRID:SCR_000306)

    Results from OddPub: Thank you for sharing your data.

    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 10.1101/2020.05.17.20104869v1 on medRxiv