GEOTranscriptomicsDanio rerioExpression profiling by high throughput sequencinghttps://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE304960GEOGSEfalseExtracellular vesicles derived from Enterococcus faecalis: inflammatory activation does not require internalizationEnterococcus 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.2026/05/27GSE304960GSM9160361GSM9160360GSM9160363GSM9160362GSM9160364GSM916035935190304960Danio rerio[42152018]