Original antigenic sin responses to Betacoronavirus spike proteins are observed in a mouse model, but are not apparent in children following SARS-CoV-2 infection

  1. Stacey A. Lapp
  2. Venkata Viswanadh Edara
  3. Austin Lu
  4. Lilin Lai
  5. Laila Hussaini
  6. Ann Chahroudi
  7. Larry J. Anderson
  8. Mehul S. Suthar
  9. Evan J. Anderson
  10. Christina A. Rostad

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Abstract

The effects of pre-existing endemic human coronavirus (HCoV) immunity on SARS-CoV-2 serologic and clinical responses are incompletely understood.

Objectives

We sought to determine the effects of prior exposure to HCoV Betacoronavirus HKU1 spike protein on serologic responses to SARS-CoV-2 spike protein after intramuscular administration in mice. We also sought to understand the baseline seroprevalence of HKU1 spike antibodies in healthy children and to measure their correlation with SARS-CoV-2 binding and neutralizing antibodies in children hospitalized with acute coronavirus disease 2019 (COVID-19) or multisystem inflammatory syndrome (MIS-C).

Methods

Groups of 5 mice were injected intramuscularly with two doses of alum-adjuvanted HKU1 spike followed by SARS-CoV-2 spike; or the reciprocal regimen of SARS-Cov-2 spike followed by HKU1 spike. Sera collected 21 days following each injection was analyzed for IgG antibodies to HKU1 spike, SARS-CoV-2 spike, and SARS-CoV-2 neutralization. Sera from children hospitalized with acute COVID-19, MIS-C or healthy controls (n = 14 per group) were analyzed for these same antibodies.

Results

Mice primed with SARS-CoV-2 spike and boosted with HKU1 spike developed high titers of SARS-CoV-2 binding and neutralizing antibodies; however, mice primed with HKU1 spike and boosted with SARS-CoV-2 spike were unable to mount neutralizing antibodies to SARS-CoV-2. HKU1 spike antibodies were detected in all children with acute COVID-19, MIS-C, and healthy controls. Although children with MIS-C had significantly higher HKU1 spike titers than healthy children (GMT 37239 vs. 7551, P = 0.012), these titers correlated positively with both SARS-CoV-2 binding (r = 0.7577, P <0.001) and neutralizing (r = 0.6201, P = 0.001) antibodies.

Conclusions

Prior murine exposure to HKU1 spike protein completely impeded the development of neutralizing antibodies to SARS-CoV-2, consistent with original antigenic sin. In contrast, the presence of HKU1 spike IgG antibodies in children with acute COVID-19 or MIS-C was not associated with diminished neutralizing antibody responses to SARS-CoV-2.

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  1. Version published to 10.1371/journal.pone.0256482
  2. ScreenIT

    SciScore for 10.1101/2021.04.29.21256344: (What is this?)

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

    Table 1: Rigor

    EthicsIACUC: All animal experiments were conducted according to approved protocols by the Emory University Institutional Animal Care and Use Committee (PROTO202000026).
    Consent: Human subjects: Children and adolescents, 0 to 21 years of age, hospitalized at Children’s Healthcare of Atlanta (CHOA) with confirmed or suspected COVID-19 or MIS-C were enrolled into an IRB-approved specimen collection protocol (Emory University IRB protocols STUDY00022371 and STUDY00000723) following written or verbal informed consent and assent as appropriate for age as previously described (12).
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Plates were washed 4 times with PBS/0.05% Tween-20, and 100 µL of horseradish peroxidase-conjugated anti-Fc IgG antibody (Jackson ImmunoResearch Laboratories, 109-035-098), diluted 1:5,000 in ELISA buffer, was added and incubated for 60 minutes at room temperature.
    anti-Fc IgG
    suggested: None
    Cells were incubated with an anti-SARS-CoV spike primary antibody directly conjugated to biotin (CR3022-biotin) for 1 hour at room temperature.
    anti-SARS-CoV spike
    suggested: None
    CR3022-biotin
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    The antibody-virus mixture was then added to Vero cells and incubated at 37°C for 1 hour.
    Vero
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    One group of five Balb/c mice was primed and boosted at 21 days IM with 10 µg SARS-CoV-2 nucleocapsid protein (SinoBiological, 40588-V08B) in 50 µl with alum (Alhydrogel adjuvant 2%, Invivogen).
    Balb/c
    suggested: None
    Software and Algorithms
    SentencesResources
    Statistical Analysis: Statistical comparisons were made with GraphPad Prism (v9.0)
    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: We detected the following sentences addressing limitations in the study:
    Limitations of this study include the small number of mice in each group, and the limited number of available clinical samples. Importantly, we also lacked pre- and post-COVID-19 sera in a single patient cohort to definitively answer the question of the effects of pre-existing HCoV antibodies on SARS-CoV-2 acquisition and clinical outcomes. Population-based studies may provide greater insights into subtler effects of pre-existing HCoV cross-reactive immunity on SARS-CoV-2 infection. We only analyzed serologic immunity to one endemic coronavirus (HKU1 in the Betacoronavirus genus), and differences could exist among Alphacoronaviruses (229E and NL63) or with the other endemic Betacoronavirus (OC43). While HKU1 infection is less prevalent than NL63 and OC43, it shares more homology in the spike protein with SARS-CoV-2 compared to the other HCoVs (20, 21), so we chose to evaluate it for this reason in these experiments. Pre-existing immunity to other SARS-CoV-2 antigens such as the nucleocapsid protein may contribute to clinical response, but we did not evaluate these antibodies in this study. The converse question of whether SARS-CoV-2 antibodies could blunt serologic responses to endemic coronaviruses or to emerging SARS-CoV-2 variants remains to be determined.

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

    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.
    • Thank you for including a protocol registration statement.

    Results from scite Reference Check: We found no unreliable references.

    About SciScore

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  3. Version published to 10.1101/2021.04.29.21256344 on medRxiv