Heetanshi Jain, Ashish Kumar, Sameh Almousa, Shalini Mishra, Kendall L. Langsten, Susy Kim, Mitu Sharma, Yixin Su, Sangeeta Singh, Bethany A. Kerr, Gagan Deep
{"title":"沿肠道-肝门静脉-肝轴的 LPS+ 细菌胞外囊泡的特征。","authors":"Heetanshi Jain, Ashish Kumar, Sameh Almousa, Shalini Mishra, Kendall L. Langsten, Susy Kim, Mitu Sharma, Yixin Su, Sangeeta Singh, Bethany A. Kerr, Gagan Deep","doi":"10.1002/jev2.12474","DOIUrl":null,"url":null,"abstract":"<p>Gut microbiome dysbiosis is a major contributing factor to several pathological conditions. However, the mechanistic understanding of the communication between gut microbiota and extra-intestinal organs remains largely elusive. Extracellular vesicles (EVs), secreted by almost every form of life, including bacteria, could play a critical role in this inter-kingdom crosstalk and are the focus of present study. Here, we present a novel approach for isolating lipopolysaccharide (LPS)+ bacterial extracellular vesicles (bEV<sup>LPS</sup>) from complex biological samples, including faeces, plasma and the liver from lean and diet-induced obese (DIO) mice. bEV<sup>LPS</sup> were extensively characterised using nanoparticle tracking analyses, immunogold labelling coupled with transmission electron microscopy, flow cytometry, super-resolution microscopy and 16S sequencing. In liver tissues, the protein expressions of TLR4 and a few macrophage-specific biomarkers were assessed by immunohistochemistry, and the gene expressions of inflammation-related cytokines and their receptors (<i>n</i> = 89 genes) were measured using a PCR array. Faecal samples from DIO mice revealed a remarkably lower concentration of total EVs but a significantly higher percentage of LPS+ EVs. Interestingly, DIO faecal bEV<sup>LPS</sup> showed a higher abundance of <i>Proteobacteria</i> by 16S sequencing. Importantly, in DIO mice, a higher number of total EVs and bEV<sup>LPS</sup> consistently entered the hepatic portal vein and subsequently reached the liver, associated with increased expression of TLR4, macrophage markers (F4/80, CD86 and CD206), cytokines and receptors (<i>Il1rn</i>, <i>Ccr1</i>, <i>Cxcl10</i>, <i>Il2rg</i> and <i>Ccr2</i>). Furthermore, a portion of bEV<sup>LPS</sup> escaped liver and entered the peripheral circulation. In conclusion, bEV could be the key mediator orchestrating various well-established biological effects induced by gut bacteria on distant organs.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":null,"pages":null},"PeriodicalIF":15.5000,"publicationDate":"2024-07-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.12474","citationCount":"0","resultStr":"{\"title\":\"Characterisation of LPS+ bacterial extracellular vesicles along the gut-hepatic portal vein-liver axis\",\"authors\":\"Heetanshi Jain, Ashish Kumar, Sameh Almousa, Shalini Mishra, Kendall L. Langsten, Susy Kim, Mitu Sharma, Yixin Su, Sangeeta Singh, Bethany A. Kerr, Gagan Deep\",\"doi\":\"10.1002/jev2.12474\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Gut microbiome dysbiosis is a major contributing factor to several pathological conditions. However, the mechanistic understanding of the communication between gut microbiota and extra-intestinal organs remains largely elusive. Extracellular vesicles (EVs), secreted by almost every form of life, including bacteria, could play a critical role in this inter-kingdom crosstalk and are the focus of present study. Here, we present a novel approach for isolating lipopolysaccharide (LPS)+ bacterial extracellular vesicles (bEV<sup>LPS</sup>) from complex biological samples, including faeces, plasma and the liver from lean and diet-induced obese (DIO) mice. bEV<sup>LPS</sup> were extensively characterised using nanoparticle tracking analyses, immunogold labelling coupled with transmission electron microscopy, flow cytometry, super-resolution microscopy and 16S sequencing. In liver tissues, the protein expressions of TLR4 and a few macrophage-specific biomarkers were assessed by immunohistochemistry, and the gene expressions of inflammation-related cytokines and their receptors (<i>n</i> = 89 genes) were measured using a PCR array. Faecal samples from DIO mice revealed a remarkably lower concentration of total EVs but a significantly higher percentage of LPS+ EVs. Interestingly, DIO faecal bEV<sup>LPS</sup> showed a higher abundance of <i>Proteobacteria</i> by 16S sequencing. Importantly, in DIO mice, a higher number of total EVs and bEV<sup>LPS</sup> consistently entered the hepatic portal vein and subsequently reached the liver, associated with increased expression of TLR4, macrophage markers (F4/80, CD86 and CD206), cytokines and receptors (<i>Il1rn</i>, <i>Ccr1</i>, <i>Cxcl10</i>, <i>Il2rg</i> and <i>Ccr2</i>). Furthermore, a portion of bEV<sup>LPS</sup> escaped liver and entered the peripheral circulation. 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Characterisation of LPS+ bacterial extracellular vesicles along the gut-hepatic portal vein-liver axis
Gut microbiome dysbiosis is a major contributing factor to several pathological conditions. However, the mechanistic understanding of the communication between gut microbiota and extra-intestinal organs remains largely elusive. Extracellular vesicles (EVs), secreted by almost every form of life, including bacteria, could play a critical role in this inter-kingdom crosstalk and are the focus of present study. Here, we present a novel approach for isolating lipopolysaccharide (LPS)+ bacterial extracellular vesicles (bEVLPS) from complex biological samples, including faeces, plasma and the liver from lean and diet-induced obese (DIO) mice. bEVLPS were extensively characterised using nanoparticle tracking analyses, immunogold labelling coupled with transmission electron microscopy, flow cytometry, super-resolution microscopy and 16S sequencing. In liver tissues, the protein expressions of TLR4 and a few macrophage-specific biomarkers were assessed by immunohistochemistry, and the gene expressions of inflammation-related cytokines and their receptors (n = 89 genes) were measured using a PCR array. Faecal samples from DIO mice revealed a remarkably lower concentration of total EVs but a significantly higher percentage of LPS+ EVs. Interestingly, DIO faecal bEVLPS showed a higher abundance of Proteobacteria by 16S sequencing. Importantly, in DIO mice, a higher number of total EVs and bEVLPS consistently entered the hepatic portal vein and subsequently reached the liver, associated with increased expression of TLR4, macrophage markers (F4/80, CD86 and CD206), cytokines and receptors (Il1rn, Ccr1, Cxcl10, Il2rg and Ccr2). Furthermore, a portion of bEVLPS escaped liver and entered the peripheral circulation. In conclusion, bEV could be the key mediator orchestrating various well-established biological effects induced by gut bacteria on distant organs.
期刊介绍:
The Journal of Extracellular Vesicles is an open access research publication that focuses on extracellular vesicles, including microvesicles, exosomes, ectosomes, and apoptotic bodies. It serves as the official journal of the International Society for Extracellular Vesicles and aims to facilitate the exchange of data, ideas, and information pertaining to the chemistry, biology, and applications of extracellular vesicles. The journal covers various aspects such as the cellular and molecular mechanisms of extracellular vesicles biogenesis, technological advancements in their isolation, quantification, and characterization, the role and function of extracellular vesicles in biology, stem cell-derived extracellular vesicles and their biology, as well as the application of extracellular vesicles for pharmacological, immunological, or genetic therapies.
The Journal of Extracellular Vesicles is widely recognized and indexed by numerous services, including Biological Abstracts, BIOSIS Previews, Chemical Abstracts Service (CAS), Current Contents/Life Sciences, Directory of Open Access Journals (DOAJ), Journal Citation Reports/Science Edition, Google Scholar, ProQuest Natural Science Collection, ProQuest SciTech Collection, SciTech Premium Collection, PubMed Central/PubMed, Science Citation Index Expanded, ScienceOpen, and Scopus.