Proteomic Analysis of Human Milk Reveals Nutritional and Immune Benefits in the Colostrum from Mothers with COVID-19

  1. Juanjuan Guo
  2. Minjie Tan
  3. Jing Zhu
  4. Ye Tian
  5. Huanyu Liu
  6. Fan Luo
  7. Jianbin Wang
  8. Yanyi Huang
  9. Yuanzhen Zhang
  10. Yuexin Yang
  11. Guanbo Wang

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Abstract

Despite the well-known benefits of breastfeeding and the World Health Organization’s breastfeeding recommendations for COVID-19 infected mothers, whether these mothers should be encouraged to breastfeed is under debate due to concern about the risk of virus transmission and lack of evidence of breastmilk’s protective effects against the virus. Here, we provide a molecular basis for the breastfeeding recommendation through mass spectrometry (MS)-based proteomics and glycosylation analysis of immune-related proteins in both colostrum and mature breastmilk collected from COVID-19 patients and healthy donors. The total protein amounts in the COVID-19 colostrum group were significantly higher than in the control group. While casein proteins in COVID-19 colostrum exhibited significantly lower abundances, immune-related proteins, especially whey proteins with antiviral properties against SARS-CoV-2, were upregulated. These proteins were detected with unique site-specific glycan structures and improved glycosylation diversity that are beneficial for recognizing epitopes and blocking viral entry. Such adaptive differences in milk from COVID-19 mothers tended to fade in mature milk from the same mothers one month postpartum. These results suggest that feeding infants colostrum from COVID-19 mothers confers both nutritional and immune benefits, and provide molecular-level insights that aid breastmilk feeding decisions in cases of active infection.

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  1. Version published to 10.3390/nu14122513
  2. ScreenIT

    SciScore for 10.1101/2022.02.25.481966: (What is this?)

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

    Table 1: Rigor

    EthicsIRB: This study was approved by the Medical Ethics Committee of Zhongnan Hospital of Wuhan University (approval no. 2020031).
    Consent: Written consent was obtained from each patient.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.

    Table 2: Resources

    Software and Algorithms
    SentencesResources
    Data processing: Following Zhu et al. (2020)19, the raw shotgun LC-MS/MS data were searched with Proteome Discoverer version 2.3 (Thermo Scientific) using the Sequest HT search engine against a UniProt Swiss-Prot database63: Homo sapiens (canonical and isoform) (October 2020; 26,566 entries).
    Proteome Discoverer
    suggested: (Proteome Discoverer, RRID:SCR_014477)
    For database searching, we used a 10-ppm precursor mass tolerance and a fragment mass tolerance of 0.02 Da followed by data filtering using Percolator, thus resulting in a 1% false discovery rate (FDR).
    Percolator
    suggested: (OMSSAPercolator, RRID:SCR_000287)
    Statistical Analysis: All statistical analyses were performed using SPSS 21.0 (SPSS Inc., Chicago, IL, USA), Perseus v.1.6.14.065, or under the R Statistical Computing Environment (v. 4.0.2).
    SPSS
    suggested: (SPSS, RRID:SCR_002865)
    Perseus
    suggested: (Perseus, RRID:SCR_015753)
    For identification of up and down-regulated proteins, a Cytoscape (v3.6.0) App ClueGOwas66 used based on the terms “biological processes” in the human database.
    Cytoscape
    suggested: (Cytoscape, RRID:SCR_003032)

    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.

    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 10.1101/2022.02.25.481966v1 on bioRxiv