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Extracellular vesicles derived from Enterococcus faecalis: inflammatory activation does not require internalization

ABSTRACT: Enterococcus faecalis is a common gut commensal Gram-positive bacterium that can act as an opportunistic pathogen and is frequently associated with nosocomial and iatrogenic bloodstream infections, such as bacteremia and endocarditis. Bacteria-derived extracellular vesicles (EVs) emerge as key mediators of host-bacteria communication with immunomodulatory roles and mechanistic participation in pathophysiological processes, such as inflammation and infection. However, the functional impact of E. faecalis-derived EVs (Ef-EVs) on host responses and their potential role in shaping host responses during infection remain unclear. In this work, we investigated the immunomodulatory effects of Ef-EVs in vitro on NF-κB/AP-1 reporter cells, primary human monocyte-derived macrophages (HMDMs), human umbilical vein endothelial cells (HUVECs), and macrophages derived from in vivo Ef-EV-treated zebrafish larvae. We found that Ef-EVs can induce pro-inflammatory responses in host macrophages via Toll-like receptor 2 (TLR2) signaling. Using bottom-up assembled bacterial EVs functionalized with the synthetic bacterial ligands as a minimalistic approach to study mechanisms of EV signaling, we demonstrate that Ef-EVs target the plasma membrane TLR2 to induce inflammation in a process uncoupled from their internalization. Furthermore, we found that Ef-EVs induce metabolic reprogramming towards a pro-inflammatory, glycolytic phenotype. Our findings showcase a previously understudied role of Gram-positive EVs in modulating immune signaling and metabolic pathways, advancing our understanding of host-pathogen communication.

ORGANISM(S): Danio rerio

PROVIDER: GSE304960 | GEO | 2026/05/27

REPOSITORIES: GEO

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Crosstalk between the effects of extracellular vesicles (Evs) and TNF on U937 human monocyte cell line

Project description:Under physiological conditions, extracellular vesicles (EVs) are present simultaneously in the extracellular compartment together with cytokines. Thus, we hypothesized that EVs in combination with cytokines induce different responses of monocyte cells compared to EVs or cytokines alone. Human monocyte U937 cells were incubated with EV-containing or EV-free CCRF human T-cell supernatant, with or without the addition of TNF. U937 cells cultured in EV-free supernatant, supernatant containing CCRF t-cell derived EVs, TNF or both. Each treatment option was measured in 3 replicates.

2013-06-19 | E-GEOD-47897 | biostudies-arrayexpress

Extracellular vesicles and their RNA cargo facilitate bidirectional cross-kingdom communication between human and bacterial cells [smallRNA-Seq]

Project description:Extracellular vesicles (EVs), released by both eukaryotic and prokaryotic cells, have emerged as key mediators of cell-to-cell communication. Recent advances highlight their crucial role in cross-kingdom communication, bridging the microbial world with human biology. Here, we investigated the molecular mechanisms underlying EV-mediated bidirectional communication within the gastrointestinal ecosystem. Using a model that includes human colon cells and both Gram-positive and Gram-negative gut bacteria, we reveal an intricate exchange of information between these kingdoms. Our analysis uncovered highly specific responses of host cells to bacterial EVs (BEVs) and BEV-RNA cargo, including uptake rates by human cells, impact on human cell viability, and alterations in their transcriptomic landscape. In parallel, we discovered that host-derived EVs and miR-192-5p are internalized by gut bacteria, leading to changes in their growth pattern. These findings highlight the precision with which EVs and their RNA cargo mediate interkingdom communication. Our results underscore the importance of tailored, context-specific analyses for understanding the scope of EV-mediated interactions in complex biological systems.

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Extracellular vesicles and their RNA cargo facilitate bidirectional cross-kingdom communication between human and bacterial cells [RNA-Seq]

2026-02-20 | GSE297395 | GEO

Microarray analysis of macrophages treated with Extracellular Vesicles derived from Neural Stem Cells

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2020-08-13 | E-MTAB-8250 | biostudies-arrayexpress

Role of miRNAs in acute kidney injury recovery induced by mesenchymal stromal cells and derived extracellular vesicles.

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Proteomic analysis of NLRP3 inflammasome-induced extracellular vesicles (infosomes) from human macrophages using LC-MS/MS

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2026-01-14 | PXD064778 | JPOST Repository

Mesenchymal stem cells exert renoprotection via extracellular vesicle-mediated modulation of M2 macrophages and spleen-kidney network.

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2022-06-21 | GSE179301 | GEO

Mapping the miRNA landscape of primitive macrophage extracellular vesicles highlights their pro-vasculogenic effects in engineered human cardiac tissue

Project description:Resident cardiac macrophages, derived from primitive yolk sac precursors during embryogenesis, have increasingly been recognized for their distinct phenotype and functions in regulating homeostasis of the human heart. However, the profile of their extracellular vesicles (EVs) in cardiac signalling and regulation remains uncharted. Here, we employ differentiation of human pluripotent stem cell (hPSC)-derived primitive macrophages (Mac) for isolation of their secreted EVs and in-depth characterization of EV-miRNA cargo. Primitive macrophages secreted nanoscale EVs that expressed canonical EV markers, and miRNA sequencing highlighted a diverse and unique profile of miRNAs when compared to EVs sourced from other principal cardiac cell lineages and published data from monocyte-derived cells. In particular, we noted the abundance and enrichment of vascular-modulatory let-7 miRNAs and miR-126-3p. Functional screening of Mac-EVs in a 3D model of in vitro cardiac vasculogenesis confirmed enhanced early endothelial cell organization and branching. Establishing a reference for the human Mac-EV miRNome enables further hypothesis-driven mechanistic tests of Mac-EV miRNAs in mediating cardiac physiology and disease, opening the door to identification of therapeutic targets and modalities for cardiac repair.

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Osteoblasts secrete miRNA-containing extracellular vesicles that enhance expansion of human umbilical cord blood cells

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2016-10-05 | E-MTAB-4560 | biostudies-arrayexpress

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