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Antimicrobial Peptides in Human Milk
Author(s) -
Filgueira Luis,
Kueffer Gwendoline K,
Twigger AleciaJane,
Walch Michael,
Geddes Donna T
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.650.24
Subject(s) - granulysin , perforin , granzyme , granzyme b , immune system , biology , flow cytometry , immunology , microbiology and biotechnology , t cell , cd8
Background Early human milk (HM) contains a large variety of immune components, including leucocytes, providing protection to the newborn infant. Peptides such as perforin, granulysin and granzymes, stored in lymphocytes work together to induce apoptosis of infected host cells and pathogens such as bacteria and parasites. Thus far their presence in HM cells has not been shown. Aims This project aims to examine the presence and localization of these immune peptides in HM cells. Methods Presence of perforin, granulysin and granzymes (A, B, H and M) was examined in cells isolated from samples of prepartum secretions (PS) collected during pregnancy and HM collected longitudinally (2–5 sampling occasions) from participants (n=31) during the first year postpartum. Gene expression was analyzed through RNA sequencing and qRT‐PCR. Linear modeling and principle component analysis between the genes was conducted. Additionally flow cytometry was performed to investigate the presence of the peptides at the protein level. Results Expression of all genes was confirmed in PS and month 1 HM samples, where variation of expression was found between women and time points. Strong correlations were found between granzyme A – perforin (r 2 =0.86), granzyme A – CD45 (r 2 =0.76), perforin – CD45 (r 2 =0.68) and perforin – granzyme B (r 2 =0.67). Principle component analysis showed a cluster of all immune peptides in addition to the biggest variation between CD45 and EPCAM. Consequently, current investigations are being undertaken to examine the co‐expression of those immune peptides in HM cells at the protein level using flow cytometry. Comparison of gene expression in a healthy and a mastitic breast from a participant showed a significant increase in CD45 and all immune peptides in the presence of mastitis. Conclusion Presence of perforin, granulysin and granzymes has been confirmed in HM cells. An increase in gene expression of these peptides has been confirmed in one participant suffering from mastitis. Further investigations are required to elucidate the roles of these immune peptides for the infant and/or mother.