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OMVs traffic outer membrane molecules, such as the porin PorB and lipo\u2010oligosaccharide (LOS), into host innate immune cells, eliciting programmed cell death pathways, and inflammation. Little is known, however, about the proteome and LOS content of OMVs released by clinical strains isolated from different infection sites, and whether these vesicles similarly activate immune responses. Here, we characterized OMVs from four <jats:italic>N. gonorrhoeae<\/jats:italic> isolates and determined their size, abundance, proteome, LOS content, and activation of inflammatory responses in macrophages. The overall proteome of the OMVs was conserved between the four different isolates, which included major outer membrane and periplasm proteins. Despite this, we observed differences in the rate of OMV biogenesis and the relative abundance of membrane proteins and LOS. Consequently, OMVs from clinical isolates induced varying rates of macrophage cell death and the secretion of interleukin\u20101 family members, such as IL\u20101\u03b1\u00a0and\u00a0IL\u20101\u03b2. Overall, these findings demonstrate that clinical isolates of <jats:italic>N. gonorrhoeae<\/jats:italic> utilize membrane vesicles to release proteins and lipids, which affects innate immune responses.<\/jats:p>","DOI":"10.1002\/pmic.202300087","type":"journal-article","created":{"date-parts":[[2023,12,7]],"date-time":"2023-12-07T23:45:11Z","timestamp":1701992711000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Characterization of outer membrane vesicles released by clinical isolates of <i>Neisseria gonorrhoeae<\/i>"],"prefix":"10.1002","volume":"24","author":[{"given":"Subhash","family":"Dhital","sequence":"first","affiliation":[{"name":"Department of Biochemistry &amp; Molecular Biology Biomedicine Discovery Institute Monash University Clayton Victoria Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pankaj","family":"Deo","sequence":"additional","affiliation":[{"name":"Department of 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Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Georg","family":"Ramm","sequence":"additional","affiliation":[{"name":"Department of Biochemistry &amp; Molecular Biology Biomedicine Discovery Institute Monash University Clayton Victoria Australia"},{"name":"Monash Ramaciotti Centre for Cryo Electron Microscopy Monash University  Melbourne Victoria Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ralf B.","family":"Schittenhelm","sequence":"additional","affiliation":[{"name":"Department of Biochemistry &amp; Molecular Biology Biomedicine Discovery Institute Monash University Clayton Victoria Australia"},{"name":"Monash Proteomics and Metabolomics Platform Biomedicine Discovery Institute Monash University  Clayton Victoria Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Benjamin","family":"Howden","sequence":"additional","affiliation":[{"name":"Microbiological Diagnostic Unit Public Health Laboratory 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