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Human breastmilk memory T cells throughout lactation manifest activated tissue-oriented profile with prominent regulation
Elise S. Saager, Arthur H. van Stigt, Butstabong Lerkvaleekul, Lisanne Lutter, Anneke H. Hellinga, M. Marlot van der Wal, Louis J. Bont, Jeanette H.W. Leusen, Belinda van’t Land, Femke van Wijk, the Protection against Respiratory tract infections through human Milk Analysis (PRIMA) group
Elise S. Saager, Arthur H. van Stigt, Butstabong Lerkvaleekul, Lisanne Lutter, Anneke H. Hellinga, M. Marlot van der Wal, Louis J. Bont, Jeanette H.W. Leusen, Belinda van’t Land, Femke van Wijk, the Protection against Respiratory tract infections through human Milk Analysis (PRIMA) group
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Research Article Immunology

Human breastmilk memory T cells throughout lactation manifest activated tissue-oriented profile with prominent regulation

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Abstract

Breastfeeding provides important immunological benefits to the neonate, but how the different immunoactive components in breastmilk contribute to immunity remains poorly understood. Here, we characterized human breastmilk T cells using single-cell RNA-Seq and flow cytometry. Breastmilk contained predominantly memory T cells, with expression of immune signaling genes, high proliferation, and an effector Th1/cytotoxic profile with high cytokine production capacities. Elevated activation was balanced by an enriched Treg population and immune regulatory markers in conventional memory T cells. Gene and surface expression of tissue-residency markers indicate that breastmilk T cells represented tissue-adapted rather than circulatory T cells. In addition, breastmilk T cells had a broad homing profile and higher activation markers in these migratory subsets. The partly overlapping transcriptome profile between breastmilk and breast tissue T cells, particularly cytotoxic T cells, might support a role in local immune defense in the mammary gland. However, unique features of breastmilk, such as Tregs, might imply an additional role in neonatal immune support. We found some correlations between the breastmilk T cell profile and clinical parameters, most notably with maternal and household factors. Together, our data suggest that breastmilk contains an adapted T cell population that exerts their function in specific tissue sites.

Authors

Elise S. Saager, Arthur H. van Stigt, Butstabong Lerkvaleekul, Lisanne Lutter, Anneke H. Hellinga, M. Marlot van der Wal, Louis J. Bont, Jeanette H.W. Leusen, Belinda van’t Land, Femke van Wijk, the Protection against Respiratory tract infections through human Milk Analysis (PRIMA) group

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Figure 1

General breastmilk T cell composition.

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General breastmilk T cell composition.
(A and B) Representative flow cyt...
(A and B) Representative flow cytometry plots of the CD4+ and CD8+ naive/memory composition based on CD45RA and CD27 staining in breastmilk- and PBMC-derived T cells. (C) Ratio of CD4+ to CD8+ T cells, excluding TCRγδ+ T cells and MAIT cells (PBMC n = 7, breastmilk [BM] 1 week n = 7, BM 1 month n = 19, BM 3 months n = 20, BM 6 months n = 4). (D and E) Distribution of naive (CD45RA+CD27+), memory (CD45RA–, split up in CD27– and CD27+) and T-EMRA (CD45RA+CD27–) T cell subsets expressed as the percentage of CD4+ (D) and CD8+ (E) T cells, with pairwise comparisons between BM time points and PBMC (PBMC n = 8, BM 1 month n = 20, BM 3 months n = 16, BM 6 months n = 12). Data represent mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 using the Kruskal-Wallis test followed by Dunn’s test for multiple comparisons. For D and E, significance only denotes differences between BM timepoints compared with control PBMC. Transparent lines connect data points of different time points postpartum from the same breastmilk donor. MAIT, Musocal-Associated Invariant T cells (TCRVα7.2+CD161+); TCR, T cell receptor. FACS data of T cells in breastmilk of 1 week and 1, 3, and 6 months postpartum compared with PBMC of age-matched female control donors.

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ISSN 2379-3708

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