Abstract

Milk small extracellular vesicles (sEV) contain proteins that provide potential information of host physiology and immunology. Bovine leukemia virus (BLV) is an oncogenic virus that causes progressive B-cell lymphosarcoma in cattle. In this study, we aimed to explore the proteomic profile of milk sEV from BLV-infected cattle compared with those from uninfected cattle. Milk sEV were isolated from three BLV-infected and three uninfected cattle. Proteomic analysis was performed by using a comprehensive nanoLC-MS/MS method. Furthermore, gene ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were used to evaluate the candidates for uniquely or differentially expressed proteins in milk sEV from BLV-infected cattle. Proteomic analysis revealed a total of 1330 common proteins in milk sEV among BLV-infected cattle, whereas 118 proteins were uniquely expressed compared with those from uninfected cattle. Twenty-six proteins in milk sEV were differentially expressed proteins more than two-fold significant difference (p < 0.05) in BLV-infected cattle. GO and KEGG analyses indicated that the candidates for uniquely or differentially expressed proteins in milk sEV had been involved in diverse biological activities including metabolic processes, cellular processes, respond to stimulus, binding, catalytic activities, cancer pathways, focal adhesion, and so on. Taken together, the present findings provided a novel insight into the proteomes of milk sEV from BLV-infected cattle.

Highlights

  • Milk small extracellular vesicles contain proteins that provide potential information of host physiology and immunology

  • Milk small extracellular vesicles (sEV) were characterized by nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), and western blot (WB) analyses

  • The results by NTA, TEM and WB analyses were indicating the typical definition and enrichment of sEV according to the MISEV2018 g­ uidelines[3]

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Summary

Introduction

Milk small extracellular vesicles (sEV) contain proteins that provide potential information of host physiology and immunology. Proteomic analysis revealed a total of 1330 common proteins in milk sEV among BLV-infected cattle, whereas 118 proteins were uniquely expressed compared with those from uninfected cattle. Milk is a diverse source of sEV that contains proteins, microRNAs (miRNAs), mRNAs, DNA, and lipids that play an important role in many biological activities including cell growth, development, immune modulation and ­regulation[4,5]. More recent study described that bovine milk sEV contained proteins, miRNAs, mRNAs, DNA, and lipids that were considered to transport biologically active cargos from donor to recipient cells for exchanging genetic i­nformation[7]. A recent study reported that milk sEV contained distinctive proteins that provided the potential information of mammary physiology of c­ attle[9]. To date, no study has reported on the proteomic analysis of milk sEV from BLV-infected cattle

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