Specific CD4+ T Cell Responses to Ancestral SARS-CoV-2 in Children Increase With Age and Show Cross-Reactivity to Beta Variant

  1. Kevin Paul
  2. Freya Sibbertsen
  3. Daniela Weiskopf
  4. Marc Lütgehetmann
  5. Madalena Barroso
  6. Marta K. Danecka
  7. Laura Glau
  8. Laura Hecher
  9. Katharina Hermann
  10. Aloisa Kohl
  11. Jun Oh
  12. Julian Schulze zur Wiesch
  13. Alessandro Sette
  14. Eva Tolosa
  15. Eik Vettorazzi
  16. Mathias Woidy
  17. Antonia Zapf
  18. Dimitra E. Zazara
  19. Thomas S. Mir
  20. Ania C. Muntau
  21. Søren W. Gersting
  22. Gabor A. Dunay

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Abstract

SARS-CoV-2 is still a major burden for global health despite effective vaccines. With the reduction of social distancing measures, infection rates are increasing in children, while data on the pediatric immune response to SARS-CoV-2 infection is still lacking. Although the typical disease course in children has been mild, emerging variants may present new challenges in this age group. Peripheral blood mononuclear cells (PBMC) from 51 convalescent children, 24 seronegative siblings from early 2020, and 51 unexposed controls were stimulated with SARS-CoV-2-derived peptide MegaPools from the ancestral and beta variants. Flow cytometric determination of activation-induced markers and secreted cytokines were used to quantify the CD4+ T cell response. The average time after infection was over 80 days. CD4+ T cell responses were detected in 61% of convalescent children and were markedly reduced in preschool children. Cross-reactive T cells for the SARS-CoV-2 beta variant were identified in 45% of cases after infection with an ancestral SARS-CoV-2 variant. The CD4+ T cell response was accompanied most predominantly by IFN-γ and Granzyme B secretion. An antiviral CD4+ T cell response was present in children after ancestral SARS-CoV-2 infection, which was reduced in the youngest age group. We detected significant cross-reactivity of CD4+ T cell responses to the more recently evolved immune-escaping beta variant. Our findings have epidemiologic relevance for children regarding novel viral variants of concern and vaccination efforts.

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  1. Version published to 10.3389/fimmu.2022.867577
  2. ScreenIT

    SciScore for 10.1101/2022.01.27.22269976: (What is this?)

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

    Table 1: Rigor

    EthicsConsent: Recruiting was conducted from May 11th until June 30th 2020 after the first infection wave, during and after the first lockdown in Germany. Parents or legal guardians provided written informed consent in all cases.
    IRB: The study was approved by the local ethical committee of Hamburg (reference number: PV7336).
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Serum antibody measurements: For screening purposes, serum samples were tested for SARS-CoV-2 specific antibodies directed against the viral nucleocapsid (IgA/IgM/IgG) using Elecsys® Anti-SARS-CoV-2 Ig assay (Roche) on the cobas e411 system (Roche).
    SARS-CoV-2
    suggested: None
    To evaluate serostatus for “common cold” coronaviruses (HCoV) and to further confirm SARS-CoV-2 serostatus, serum antibodies (IgG) against the viral nucleocapsid of HCoV strains 229E, NL63, OC43, HKU1 and SARS-CoV-2 anti - S1 subunit, anti - receptor binding domain as well as anti - nucleocapsid were measured by using the recomLine SARS-CoV-2 IgG® assay (Mikrogen).
    NL63
    suggested: (Virostat Cat# 3879, RRID:AB_2889994)
    anti - S1 subunit, anti - receptor binding domain as well as anti -
    suggested: None
    anti
    suggested: None
    Expression of activation-induced markers (CD69 and OX40) in response to specific peptide stimulation, as well as their memory phenotype were measured by flow cytometry (flow cytometry antibodies are listed in Supplementary Table 1).
    CD69
    suggested: None
    OX40
    suggested: None
    For compensation, Anti-Mouse or Anti-Rat Ig, κ/Negative Control Compensation Particles Set (BD Biosciences) was used for antibodies, and ArC™ Amine Reactive Compensation Beads (Invitrogen) were used for the Dead Cell Stain Kit.
    Anti-Mouse
    suggested: None
    Anti-Rat Ig,
    suggested: None
    Software and Algorithms
    SentencesResources
    Data Analysis and Statistics: Data analysis, graphs and statistics were prepared with FlowJo version 10, and R 4.0.5 (packages: tidyverse, rstatix, splines, emmeans, kableExtra, magrittr, heatmaply).
    FlowJo
    suggested: (FlowJo, RRID:SCR_008520)

    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:
    Our study had several limitations. As children were recruited weeks or months after infection, no PCR based confirmation from the acute phase of infection was available. As previous infection was defined through seropositivity alone, a combined positivity in three independent serological tests was required and this approach should minimize the possibility of false positives. A further limitation based on the retrospective study design is that a detailed history from the time of infection as well as the exact time of infection could only be collected for a fraction of participants. Because of a reluctance in small children and their caregivers, as well as the technical difficulty regarding blood draws, younger age groups were relatively underrepresented. Also, CD8+ T cell responses were not investigated - these could provide important insights into cellular anti-SARS-CoV-2 immunity and should be the focus of further studies.

    Results from TrialIdentifier: We found the following clinical trial numbers in your paper:

    IdentifierStatusTitle
    NCT04534608Active, not recruitingCOVID-19 Child Health Investigation of Latent Disease in Ham…

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: Please consider improving the rainbow (“jet”) colormap(s) used on pages 32, 33 and 34. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.

    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.

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

    About SciScore

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  3. Version published to 10.1101/2022.01.27.22269976v1 on medRxiv