Ávila Gabriela, Bonnet Muriel, Viala Didier, Dejean Sebastian, Agazzi Alessandro, Lecchi Cristina, Ceciliani Fabrizio
Department of Veterinary and Animal Sciences, Università Degli Studi di Milano, Via dell'Università 6, 26900, Lodi, Italy.
INRAE, University of Clermont Auvergne, Vetagro Sup, UMRH, 63122, Saint-Genès-Champanelle, France.
Sci Rep. 2025 Mar 8;15(1):8069. doi: 10.1038/s41598-025-92550-3.
Small extracellular vesicles (EVs) are a subtype of nano-sized extracellular vesicles that mediate intercellular communication. EVs can be found in different body fluids, including milk. Monocytes internalize porcine milk EVs and modulate immune functions in vitro by decreasing their phagocytosis and chemotaxis while increasing their oxidative burst. This study aimed to assess the impact of porcine milk EVs on the porcine peripheral blood mononuclear cells (PBMC) proteome. Porcine PBMC were incubated with porcine milk EVs or medium as a control. Extracted proteins were then analyzed using nano-LC-MS/MS. A total of 1584 proteins were identified. The supervised multivariate statistical analysis, sparse variant partial least squares - discriminant analysis (sPLS-DA) for paired data identified discriminant proteins (DP) that contributed to a clear separation between the porcine milk EVs treated cells and control groups. A total of 384 DP from both components were selected. Gene Ontology (GO) enrichment analysis with ProteINSIDE provided the evidence that the DP with a higher abundance in porcine milk EVs, like TLR2, APOE, CD36, MFGE8, were mainly involved in innate immunity and the process of EVs uptake processes. These results provide a proteomics background to the immunomodulatory activity of porcine milk EVs and to the potential mechanisms used by immune cells to internalize them.
小细胞外囊泡（EVs）是纳米级细胞外囊泡的一种亚型，介导细胞间通讯。EVs存在于包括乳汁在内的不同体液中。单核细胞可摄取猪乳EVs，并在体外通过降低其吞噬作用和趋化性，同时增强其氧化爆发来调节免疫功能。本研究旨在评估猪乳EVs对猪外周血单个核细胞（PBMC）蛋白质组的影响。将猪PBMC与猪乳EVs或作为对照的培养基一起孵育。然后使用纳升液相色谱-串联质谱（nano-LC-MS/MS）分析提取的蛋白质。共鉴定出1584种蛋白质。对配对数据进行的监督多变量统计分析，即稀疏变量偏最小二乘判别分析（sPLS-DA），确定了有助于猪乳EVs处理细胞与对照组之间明显区分的判别蛋白（DP）。从两个组分中总共选择了384个DP。使用ProteINSIDE进行的基因本体论（GO）富集分析提供了证据，表明在猪乳EVs中丰度较高的DP，如TLR2、APOE、CD36、MFGE8，主要参与先天免疫和EVs摄取过程。这些结果为猪乳EVs的免疫调节活性以及免疫细胞摄取它们的潜在机制提供了蛋白质组学背景。
