{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T11:09:47Z","timestamp":1786446587760,"version":"build-2736575974"},"reference-count":51,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,8,4]],"date-time":"2025-08-04T00:00:00Z","timestamp":1754265600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Extracellular vesicles (EVs) derived from <jats:italic>Staphylococcus aureus<\/jats:italic> (<jats:italic>S. aureus<\/jats:italic>) carry multiple components, such as toxins, antigens, and resistance determinants, whose production is influenced by exposure to \u03b2-lactam antibiotics. However, systematic studies on the effects of \u03b2-lactam antibiotics on the release and functions of EVs remain limited.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Extracellular vesicles (EVs) were isolated from the OS-MRSA (OS-200) strain by ultracentrifugation, including both EVs untreated with antibiotics and those exposed to sub-inhibitory concentrations of oxacillin (OI-EVs). Proteomic analysis was performed using liquid chromatography-tandem mass spectrometry (LC-MS\/MS). The functional changes of these EVs were further assessed through erythrocyte hemolysis assays, biofilm formation assays, measurement of cytokines, and cell invasion assays.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Exposure to sub-inhibitory concentrations of oxacillin (half and one-eighth of the minimum inhibitory concentration [MIC]) significantly enhanced the secretion of OI-EVs and increased the abundance of several proteins, including penicillin-binding protein 2, EmrB, Hlg, and enolase, in a concentration-dependent manner. Notably, EV1\/2MIC exhibited more pronounced functional changes and enhanced hemolytic activity against rabbit red blood cells. Also, EV1\/2MIC exhibited significant promoting effects on biofilm formation of OS-200. Additionally, OI-EVs stimulated the secretion of interleukin-6 and tumor necrosis factor-\u03b1 by THP-1 macrophages in a dose-dependent manner and demonstrated greater penetration potential into A549 lung epithelial cells.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Exposure to sub-inhibitory concentrations of oxacillin significantly altered the secretion and composition of EVs, highlighting a novel relationship between antimicrobial exposure, exosome biogenesis, and OS-MRSA pathogenicity. These findings provide new insights into how \u03b2-lactam antibiotics influence host\u2013pathogen interactions by modulating the secretion of bacterial vesicles.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fmicb.2025.1616536","type":"journal-article","created":{"date-parts":[[2025,8,4]],"date-time":"2025-08-04T06:16:00Z","timestamp":1754288160000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":5,"title":["Sub-inhibitory concentrations of oxacillin modulate biogenesis and function of extracellular vesicles secreted by oxacillin-sensitive methicillin-resistant Staphylococcus aureus"],"prefix":"10.3389","volume":"16","author":[{"given":"Zhaoxia","family":"Ran","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Minghui","family":"Hao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Binxin","family":"Guo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junrui","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,8,4]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"1075","DOI":"10.1007\/s00203-018-1519-x","article-title":"The transmembrane domain of the Staphylococcus aureus ESAT-6 component EssB mediates interaction with the integral membrane protein EsaA, facilitating partially regulated secretion in a heterologous host","volume":"200","author":"Ahmed","year":"2018","journal-title":"Arch. 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