Characterization of SARS CoV-2 Antibodies in Breast Milk from 21 Women with Confirmed COVID-19 Infection
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Abstract
One potential mechanism for protection from SARS-CoV-2 in infants and young children is through passive immunity via breast milk from a mother previously infected with the novel coronavirus. The primary objectives of this study were to establish the presence of SARS-CoV-2 specific IgA and IgG and to characterize the specific antigenic regions of SARS-CoV-2 proteins that were reactive with antibodies in breast milk from women with confirmed SARS-CoV-2 infection.
Between March 2020 and September 2020, 21 women with confirmed SARS-CoV-2 infection were enrolled into Mommy’s Milk at the University of California, San Diego. Women donated serial breast milk samples. Breast milk samples were used to probe a multi-coronavirus protein microarray containing full-length proteins and variable length overlapping fragments of SARS-CoV-2 spike (S), envelope (E), membrane (M), nucleocapsid (N), and open reading frame (ORF) proteins.
The breast milk samples contained IgA reactive with a variety of SARS-CoV-2 antigens. The most IgA-reactive SARS-CoV-2 proteins were N (42.9% of women responded to 1 ≥ N fragment) and S proteins (23.9% of women responded to ≥ 1 fragment of S1 or S2). Overall, individual COVID-19 cases had diverse and unique milk IgA profiles over the course of follow-up since onset of SARS-CoV-2 symptoms.
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SciScore for 10.1101/2021.07.19.21260661: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IRB: Study Approval: The University of California San Diego Institutional Review Board approved the study, and women provided oral and written informed consent prior to use of their breast milk samples in the analyses described above.
Consent: Study Approval: The University of California San Diego Institutional Review Board approved the study, and women provided oral and written informed consent prior to use of their breast milk samples in the analyses described above.Sex as a biological variable Collection: Breast milk samples and clinical information were obtained from women participating in the Mommy’s Milk Human Milk Biorepository at the University of California, San Diego. Randomization no… SciScore for 10.1101/2021.07.19.21260661: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IRB: Study Approval: The University of California San Diego Institutional Review Board approved the study, and women provided oral and written informed consent prior to use of their breast milk samples in the analyses described above.
Consent: Study Approval: The University of California San Diego Institutional Review Board approved the study, and women provided oral and written informed consent prior to use of their breast milk samples in the analyses described above.Sex as a biological variable Collection: Breast milk samples and clinical information were obtained from women participating in the Mommy’s Milk Human Milk Biorepository at the University of California, San Diego. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Coronavirus Panel 2 multiplex electrochemiluminescence (ECLIA) serology kits were purchased from Meso Scale Discovery (Rockville, MD) to measure Immunoglobulin A and G (IgA and IgG) antibodies against four antigens related to SARS-CoV-2 in whole breast milk samples. IgGsuggested: NonePlates were incubated with 150 µL of 1x detection antibody solution containing SULFO-TAG anti-human IgA or IgG prepared with Diluent 100. anti-human IgAsuggested: NoneClinical variables were associated with SARS-CoV-2-specific IgA antibodies using multivariable linear mixed effects regression (LMER) to model antibody responses to each individual SARS-CoV-2 protein or fragment with random intercepts at the subject level to adjust for repeated measures. SARS-CoV-2-specific IgAsuggested: NoneExperimental Models: Cell Lines Sentences Resources Protein microarray analysis of breast milk samples: The first generation multi-coronavirus protein microarray, produced by Antigen Discovery, Inc. (ADI, Irvine, CA, USA), included 935 full-length coronavirus proteins, overlapping 100, 50 and 30 aa protein fragments and overlapping 13-20 aa peptides from SARS-CoV-2 (WA-1), SARS-CoV, MERS-CoV, HCoV-NL63 and HCoV-OC43. HCoV-NL63suggested: RRID:CVCL_RW88)SARS-CoV-2 and SARS-CoV S proteins were made in Sf9 insect cells and the SARS-CoV-2 RBD, made in HEK-293 cells. HEK-293suggested: CLS Cat# 300192/p777_HEK293, RRID:CVCL_0045)Software and Algorithms Sentences Resources Data visualization was performed using the circlize, ComplexHeatmap and ggplot2 packages in R (23, 24). circlizesuggested: (circlize, RRID:SCR_002141)ComplexHeatmapsuggested: (ComplexHeatmap, RRID:SCR_017270)ggplot2suggested: (ggplot2, RRID:SCR_014601)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:This study had several limitations as well as strengths. The collection of breast milk samples were not directly observed and samples were collected with nonstandard sampling time points. Therefore, the breast milk samples collected at the onset of symptoms may not reflect duration of exposure to SARS-CoV-2, which may be attributed for the differences observed in IgA kinetics. We also relied on the maternal report of SARS-CoV-2 test results, symptoms and treatments received, however, all participants completed a semi-structured interview guided by trained study staff who prompted for specifics with the aid of a calendar. Another limitation in our study was lack of reactivity to the S1 protein and its fragments produced in vitro using an E. coli based reaction mixture. This was likely due to the lack of eukaryotic post-translational modifications, including N-linked glycosylation which is abundant in S1. We compensated for this by including purified S protein and RBD in the array and by assaying purified S, RBD and the NTD of S by electro-chemiluminescent immunoassay (ECLIA). The added value of this study comes from the longitudinal assessment of milk antibody levels and the breadth of antigens covered by the protein microarray and ECLIA platforms, coupled with COVID-19 patient cases with unique profiles.
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.
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