Abstract

ScopeExtracellular vesicles, including exosomes, have been identified in all biological fluids and rediscovered as an important part of the intercellular communication. Breast milk also contains extracellular vesicles and the proposed biological function is to enhance the antimicrobial defense in newborns. It is, however, unknown whether extracellular vesicles are still present in commercial milk and, more importantly, whether they retained their bioactivity. Here, we characterize the extracellular vesicles present in semi-skimmed cow milk available for consumers and study their effect on T cells.Methods and ResultsExtracellular vesicles from commercial milk were isolated and characterized. Milk-derived extracellular vesicles contained several immunomodulating miRNAs and membrane protein CD63, characteristics of exosomes. In contrast to RAW 267.4 derived extracellular vesicles the milk-derived extracellular vesicles were extremely stable under degrading conditions, including low pH, boiling and freezing. Milk-derived extracellular vesicles were easily taken up by murine macrophages in vitro. Furthermore, we found that they can facilitate T cell differentiation towards the pathogenic Th17 lineage. Using a (CAGA)12-luc reporter assay we showed that these extracellular vesicles carried bioactive TGF-β, and that anti-TGF-β antibodies blocked Th17 differentiation.ConclusionOur findings show that commercial milk contains stable extracellular vesicles, including exosomes, and carry immunoregulatory cargo. These data suggest that the extracellular vesicles present in commercial cow milk remains intact in the gastrointestinal tract and exert an immunoregulatory effect.

Highlights

  • Extracellular vesicles (EV), including exosomes, are membrane vesicles secreted by a variety of cells and are heterogeneous in size, ranging from 30–1000nm in diameter [1]

  • Extracellular vesicles from commercial milk were isolated and characterized

  • Using a (CAGA)12-luc reporter assay we showed that these extracellular vesicles carried bioactive TGF-β, and that anti-TGF-β antibodies blocked T-helper17 cells (Th17) differentiation

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Summary

Introduction

Extracellular vesicles (EV), including exosomes, are membrane vesicles secreted by a variety of cells and are heterogeneous in size, ranging from 30–1000nm in diameter [1]. The production of EVs has been reported for many different cell types including macrophages, lymphocytes, dendritic cells, as well as epithelial and tumor cells [3,4,5,6,7,8]. EVs have been identified in a number of biological fluids such as blood, saliva and urine [9,10,11,12,13,14]. Recent studies revealed that EVs are involved in the genetic exchange of RNA and microRNAs (miRNAs) between cells [15,16,17]. Exosome-like EVs have been identified in both human breast milk and bovine colostrum [20,21]. It has been suggested that these breast milk-derived EVs are taken up systemically by the milk recipient, where they can play a role in the development of the infants immune system [22]

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