Immune and pathophysiologic profiling of antenatal coronavirus disease 2019 in the GIFT cohort: A Singaporean case-control study

  1. Yue Gu
  2. Jia Ming Low
  3. Jolene Su Yi Tan
  4. Melissa Shu Feng Ng
  5. Lisa F. P. Ng
  6. Bhuvaneshwari Shunmuganathan
  7. Rashi Gupta
  8. Paul A. MacAry
  9. Zubair Amin
  10. Le Ye Lee
  11. Derrick Lian
  12. Lynette Pei-Chi Shek
  13. Youjia Zhong
  14. Liang Wei Wang

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Abstract

COVID-19 can be severe in pregnant women, and have adverse consequences for the subsequent infant. We profiled the post-infectious immune responses in maternal and child blood as well as breast milk in terms of antibody and cytokine expression and performed histopathological studies on placentae obtained from mothers convalescent from antenatal COVID-19. Seventeen mother-child dyads (8 cases of antenatal COVID-19 and 9 healthy unrelated controls; 34 individuals in total) were recruited to the Gestational Immunity For Transfer (GIFT) study. Maternal and infant blood, and breast milk samples were collected over the first year of life. All samples were analyzed for IgG and IgA against whole SARS-CoV-2 spike protein, the spike receptor-binding domain (RBD), and previously reported immunodominant epitopes, as well as cytokine levels. The placentae were examined microscopically. The study is registered at clinicaltrials.gov under the identifier NCT04802278. We found high levels of virus-specific IgG in convalescent mothers and similarly elevated titers in newborn children. Thus, antenatal SARS-CoV-2 infection led to high plasma titers of virus-specific antibodies in infants postnatally. However, this waned within 3–6 months of life. Virus neutralization by plasma was not uniformly achieved, and the presence of antibodies targeting known immunodominant epitopes did not assure neutralization. Virus-specific IgA levels were variable among convalescent individuals’ sera and breast milk. Antibody transfer ratios and the decay of transplacentally transferred virus-specific antibodies in neonatal circulation resembled that for other pathogens. Convalescent mothers showed signs of chronic inflammation marked by persistently elevated IL17RA levels in their blood. Four placentae presented signs of acute inflammation, particularly in the subchorionic region, marked by neutrophil infiltration even though > 50 days had elapsed between virus clearance and delivery. Administration of a single dose of BNT162b2 mRNA vaccine to mothers convalescent from antenatal COVID-19 increased virus-specific IgG and IgA titers in breast milk, highlighting the importance of receiving the vaccine even after natural infection with the added benefit of enhanced passive immunity.

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  1. Version published to 10.3389/fped.2022.949756
  2. ScreenIT

    SciScore for 10.1101/2022.04.19.22273864: (What is this?)

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

    Table 1: Rigor

    EthicsIRB: Ethics statement: This study was approved by the National Healthcare Group Institutional Review Board (Gestational Immunity For Transfer GIFT: DSRB Reference Number: 2020/00483).
    Consent: Written informed consent was obtained from all subjects (and where applicable, parents), and the study was conducted in accordance with the Helsinki Declaration.
    Sex as a biological variableMultiplex microbead-based immunoassay: Quantification of cytokine levels in the plasma samples of convalescent and healthy mothers was performed by multiplex microbead-based immunoassays.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Synthesis of the SARS-CoV-2 receptor binding domain (RBD) and spike protein: SARS-CoV-2 spike and RBD were generously provided by the Antibody Engineering Programme, Life Sciences Institute, NUS as described previously [10].
    SARS-CoV-2 receptor binding domain (RBD)
    suggested: None
    Plate was then washed three times with PBST followed by 1-hour incubation in the dark with 100 µL of 5000-times diluted goat anti-human IgG-HRP (Invitrogen, #31413), or 5000-times diluted F(ab’)2 anti-human IgA-HRP (Invitrogen,
    anti-human IgG-HRP
    suggested: None
    Goat HRP-conjugated anti-human IgA (Abcam, #ab97215) diluted at 1: 2,000 in the blocking buffer was used for the detection of peptide specific antibodies.
    Goat HRP-conjugated anti-human IgA
    suggested: (Creative Diagnostics Cat# DPAB4111, RRID:AB_2393981)
    anti-human IgA
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Briefly, the ACE2 stably expressed CHO cells were cultured at 5 × 104/mL cells in complete medium for 24 hours.
    CHO
    suggested: None
    Software and Algorithms
    SentencesResources
    Ethics statement: This study was approved by the National Healthcare Group Institutional Review Board (Gestational Immunity For Transfer GIFT: DSRB Reference Number: 2020/00483).
    National Healthcare
    suggested: None
    Data analyses were performed using GraphPad Prism (GraphPad Software, version 7.0.0).
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    GraphPad
    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:
    A key limitation of our study is the cohort size. Most of our samples were collected early on in the pandemic before Singapore introduced a slew of effective public health measures to stymie the spread of the virus, including travel restrictions, social distancing, and a lockdown. Due to this unique situation, relatively few people were infected in the first wave from April to August 2020. Limited community transmission between August 2020 and August 2021 prevented further subject recruitment and sample collection. In the context of the present situation, we note two caveats, namely the current dominance of Delta and Omicron over early pandemic variants and the high rate of vaccine uptake by the resident population (∼92% as of 31st March 2022). These two factors preclude meaningful head-to-head comparisons of more recent studies against our cohort in the GIFT study where immunologically naïve individuals were infected with variants possessing lower immunoevasive abilities. Notwithstanding these differences, our studies provide insights into the pathogenesis of COVID-19 in the under-studied demographics of pregnant and lactating women as well as infants born to them.

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

    IdentifierStatusTitle
    NCT04802278RecruitingGestational Immunity for Transfer

    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.

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

    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 10.1101/2022.04.19.22273864v1 on medRxiv