Mecocci Samanta, De Paolis Livia, Zoccola Roberto, Fruscione Floriana, De Ciucis Chiara Grazia, Chiaradia Elisabetta, Moccia Valentina, Tognoloni Alessia, Pascucci Luisa, Zoppi Simona, Zappulli Valentina, Chillemi Giovanni, Goria Maria, Cappelli Katia, Razzuoli Elisabetta
Department of Veterinary Medicine, University of Perugia, 06123 Perugia, Italy.
National Reference Center of Veterinary and Comparative Oncology (CEROVEC), Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d'Aosta, Piazza Borgo Pila 39-24, 16129 Genova, Italy.
Biomedicines. 2022 Dec 15;10(12):3264. doi: 10.3390/biomedicines10123264.
Extracellular Vesicles (EVs) are nano-sized double-lipid-membrane-bound structures, acting mainly as signalling mediators between distant cells and, in particular, modulating the immune response and inflammation of targeted cells. Milk and colostrum contain high amounts of EVs that could be exploited as alternative natural systems in antimicrobial fighting. The aim of this study is to evaluate cow colostrum-derived EVs (colosEVs) for their antimicrobial, anti-inflammatory and immunomodulating effects in vitro to assess their suitability as natural antimicrobial agents as a strategy to cope with the drug resistance problem. ColosEVs were evaluated on a model of neonatal calf diarrhoea caused by Escherichia coli infection, a livestock disease where antibiotic therapy often has poor results. Colostrum from Piedmontese cows was collected within 24 h of calving and colosEVs were immediately isolated. IPEC-J2 cell line was pre-treated with colosEVs for 48 h and then infected with EPEC/NTEC field strains for 2 h. Bacterial adherence and IPEC-J2 gene expression analysis (RT-qPCR) of CXCL8, DEFB1, DEFB4A, TLR4, TLR5, NFKB1, MYD88, CGAS, RIGI and STING were evaluated. The colosEVs pre-treatment significantly reduced the ability of EPEC/NTEC strains to adhere to cell surfaces (p = 0.006), suggesting a role of ColosEVs in modulating host−pathogen interactions. Moreover, our results showed a significant decrease in TLR5 (p < 0.05), CGAS (p < 0.05) and STING (p < 0.01) gene expression in cells that were pre-treated with ColosEVs and then infected, thus highlighting a potential antimicrobial activity of ColosEVs. This is the first preliminarily study investigating ColosEV immunomodulatory and anti-inflammatory effects on an in vitro model of neonatal calf diarrhoea, showing its potential as a therapeutic and prophylactic tool.
细胞外囊泡（EVs）是纳米级的双脂质膜结合结构，主要作为远距离细胞之间的信号介质，尤其能调节靶细胞的免疫反应和炎症。牛奶和初乳中含有大量的细胞外囊泡，可作为抗菌斗争中的替代天然系统加以利用。本研究的目的是评估牛初乳来源的细胞外囊泡（初乳细胞外囊泡）在体外的抗菌、抗炎和免疫调节作用，以评估其作为天然抗菌剂应对耐药性问题的适用性。在由大肠杆菌感染引起的新生小牛腹泻模型上评估了初乳细胞外囊泡，这是一种抗生素治疗效果往往不佳的家畜疾病。在产犊后24小时内收集皮埃蒙特奶牛的初乳，并立即分离初乳细胞外囊泡。将IPEC-J2细胞系用初乳细胞外囊泡预处理48小时，然后用肠致病性大肠杆菌/非典型肠产毒性大肠杆菌田间菌株感染2小时。评估细菌黏附以及CXCL8、DEFB1、DEFB4A、TLR4、TLR5、NFKB1、MYD88、CGAS、RIGI和STING的IPEC-J2基因表达分析（RT-qPCR）。初乳细胞外囊泡预处理显著降低了肠致病性大肠杆菌/非典型肠产毒性大肠杆菌菌株黏附细胞表面的能力（p = 0.006），表明初乳细胞外囊泡在调节宿主-病原体相互作用中发挥作用。此外，我们的结果显示，在先用初乳细胞外囊泡预处理然后感染的细胞中，TLR5（p < 0.05）、CGAS（p < 0.05）和STING（p < 0.01）基因表达显著降低，从而突出了初乳细胞外囊泡的潜在抗菌活性。这是第一项初步研究，调查初乳细胞外囊泡对新生小牛腹泻体外模型的免疫调节和抗炎作用，显示了其作为治疗和预防工具的潜力。
