Humoral and Cell-Mediated Immune Response in Colostrum from Women Diagnosed Positive for SARS-CoV-2

  1. Vignesh Narayanaswamy
  2. Brian Pentecost
  3. Dominique Alfandari
  4. Emily Chin
  5. Kathleen Minor
  6. Alyssa Kastrinakis
  7. Tanya Lieberman
  8. Kathleen F. Arcaro
  9. Heidi Leftwich

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Abstract

To evaluate the immune response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in colostrum from women who tested positive for the virus.

Methods:

Between March and September 2020 we obtained bilateral colostrum samples collected on spot cards within 48 hours of delivery from 15 new mothers who had previously tested positive for SARS-CoV-2. Four of 15 women provided liquid colostrum, which was used for validating results obtained from spot cards. Archived bilateral colostrum samples collected from 8 women during 2011–2013 were used as pre-coronavirus disease 2019 (COVID-19) controls. All samples were tested for reactivity to the receptor binding domain (RBD) of the SARS-CoV-2 spike protein using an enzyme-linked immunosorbent assay that measures SARS-CoV-2 RBD-specific IgA, IgG, and IgM and for levels of 10 inflammatory cytokines (interferon-gamma [IFN-γ], tumor necrosis factor-alpha, interleukin [IL]-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-13) using a multiplex electrochemiluminescent sandwich assay.

Results:

Our validation studies indicate that the levels of SARS-CoV-2-specific antibodies and the associated cytokines measured in liquid colostrum are comparable to levels eluted from spot cards. Bilateral colostrum samples from 73%, 73%, and 33% of the 15 COVID-19 mothers exhibited IgA, IgG, and IgM reactivity to RBD, respectively. In addition, symptomatic COVID-19 mothers had statistically significant elevated levels of 4 of the 10 inflammatory markers (IFN-γ, IL-4, IL-6, and IL-12) compared to asymptomatic COVID-19 mothers.

Conclusions:

A strong humoral immune response is present in the colostrum of women who were infected with SARS-CoV-2 before delivering. The evolution and duration of the antibody response, as well as dynamics of the cytokine response, remain to be determined. Our results also indicate that future large-scale studies can be conducted with milk easily collected on paper spot cards.

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  1. Version published to 10.1089/bfm.2021.0082
  2. ScreenIT

    SciScore for 10.1101/2021.01.03.20248715: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementConsent: Recruitment of COVID-19-positive participants: Study participants were patients at UMass Memorial Medical Center (UMMC, Worcester, MA) and provided consent in accordance with an IRB-approved protocol (H00020140).
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variableSelection of pre-COVID-19 controls: We identified archived samples from eight women who donated liquid bilateral colostrum (1-3 days post-partum) during June 2011-May 2013.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    The receptor binding domain (RBD) spike protein cloned into the pCAGGS expression vector was expressed in HEK293T cells (ATCC) using PEI (10:1 PEI:DNA ratio) and purified by gravity flow, as described in Stadlbauer et al19.
    HEK293T
    suggested: None
    Software and Algorithms
    SentencesResources
    Briefly, 96-well plates (Fisher Sci., #351172) were coated with the RBD spike protein at 1 μg/mL in 1X phosphate-buffered saline and incubated with gentle shaking overnight at 4°C, followed by blocking in 5% (w/v) dry skimmed milk in TBST with gentle shaking for 30 minutes at RT.
    Fisher Sci.
    suggested: (One Mind Biospecimen Bank Listing, RRID:SCR_004193)
    Plates were washed 3 times, incubated with 2,2’-Azinobis [3-ethylbenzothiazoline-6-sulfonic acid]-diammonium salt (ATBS; Sigma Aldrich, #A9941) diluted at 0.2 mg/mL in 0.1 M Sodium Acetate pH 4.5 at 370C for 30 minutes.
    ATBS; Sigma Aldrich
    suggested: None

    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.

  3. Version published to 10.1101/2021.01.03.20248715 on medRxiv