Background:Lactobacillus reuteri strains are widely used as probiotics to prevent and treat inflammatory bowel disease by modulating the host’s immune system.However,the underlying mechanisms by which they communicate with the host have not been clearly understood.Bacterial extracellular vesicles(EVs)have been considered as important mediators of host-pathogen interactions,but their potential role in commensals-host crosstalk has not been widely studied.Here,we investigated the regulatory actions of EVs produced by L.reuteri BBC3,a gut-associated commensal bacterium of Black-Bone chicken,in the development of lipopolysaccharide(LPS)-induced intestinal inflammation in a chicken model using both in vivo and in vitro experiments.Results:L.reuteri BBC3 produced nano-scale membrane vesicles with the size range of 60-250 nm.Biochemical and proteomic analyses showed that L.reuteri BBC3-derived EVs(LrEVs)carried DNA,RNA and several bioactive proteins previously described as mediators of other probiotics’beneficial effects such as glucosyltransferase,serine protease and elongation factor Tu.In vivo broiler experiments showed that administration of LrEVs exerted similar effects as L.reuteri BBC3 in attenuating LPS-induced inflammation by improving growth performance,reducing mortality and decreasing intestinal injury.LrEVs suppressed the LPS-induced expression of pro-inflammatory genes(TNF-α,IL-1β,IL-6,IL-17 and IL-8),and improved the expression of anti-inflammatory genes(IL-10 and TGF-β)in the jejunum.LrEVs could be internalized by chicken macrophages.In vitro pretreatment with LrEVs reduced the gene expression of TNF-α,IL-1βand IL-6 by suppressing the NF-κB activity,and enhanced the gene expression of IL-10 and TGF-βin LPS-activated chicken macrophages.Additionally,LrEVs could inhibit Th1-and Th17-mediated inflammatory responses and enhance the immunoregulatory cells-mediated immunosuppression in splenic lymphocytes of LPS-challenged chickens through the activation of macrophages.Finally,we revealed that the r
Rujiu HuHua LinMimi WangYuezhen ZhaoHaojing LiuYuna MinXiaojun YangYupeng GaoMingming Yang
