{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,5,27]],"date-time":"2026-05-27T01:06:48Z","timestamp":1779844008665,"version":"3.53.1"},"posted":{"date-parts":[[2026,5,22]]},"group-title":"Microbiology","reference-count":36,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,5,22]],"date-time":"2026-05-22T00:00:00Z","timestamp":1779408000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"name":"National Institute of Child Health and Human Development","award":["R01HD102318"],"award-info":[{"award-number":["R01HD102318"]}]},{"name":"National Institute of Child Health and Human Development","award":["R01HD098867"],"award-info":[{"award-number":["R01HD098867"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,5,22]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>\n                    The female lower reproductive tract harbors a complex microbiome that plays a critical role in reproductive health. A vaginal microbiome dominated by\n                    <jats:italic>Lactobacillus crispatus<\/jats:italic>\n                    (LC; Community State Type (CST) I) supports vaginal health, whereas a microbiome enriched with anaerobic species, such as\n                    <jats:italic>Gardnerella vaginalis<\/jats:italic>\n                    (GV) and\n                    <jats:italic>Mobiluncus mulieris<\/jats:italic>\n                    (MM) (CST IV) is linked to bacterial vaginosis (BV) and adverse outcomes, including sexually transmitted infections, infertility, and preterm birth. Although antibiotics such as metronidazole and clindamycin are commonly prescribed to treat BV, recurrence rates remain high, and the impact of these treatments on bacterial extracellular vesicles (bEVs), critical mediators of host-microbe interactions, is poorly understood.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Result<\/jats:title>\n                  <jats:p>We investigated how antibiotic treatment at a dose below minimum inhibitory concentration alters the production and immunomodulatory function of bEVs derived from GV, MM, and LC. Using nanoparticle tracking analysis, cytokine profiling, and TLR pathway analyses, we found that antibiotic treatment significantly enhanced the inflammatory properties of bEVs in a species- and antibiotic-specific manner. Notably, bEVs from antibiotic-exposed GV and MM cultures induced elevated cytokine responses in epithelial and immune cells, primarily through TLR2 activation for GV bEVs, and through both TLR2 and TLR5 activation for MM bEVs. While LC bEVs are typically non-inflammatory, exposure to metronidazole, even at a lower dose than what is used clinically, rendered them immunostimulatory, suggesting a potential unintended proinflammatory consequence of treatment on beneficial microbes. We also detected bEVs in human vaginal swabs, including vaginolysin-positive bEVs, even in CST I microbiomes, indicating that low-abundance microbes, including pathogens, remain transcriptionally active.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>These findings suggest that antibiotics not only reduce microbial load but also reshape bacterial communication via bEVs, potentially contributing to inflammation, epithelial barrier disruption, persistent dysbiosis, and recurrent infections. This work underscores the need for precision antimicrobial strategies that eliminate pathogens while preserving beneficial bacteria and their functional bEVs. Future therapies may benefit from considering the ecosystem-wide effects of antibiotics on the vaginal microbiome and its bEV-mediated signaling network.<\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.05.21.726874","type":"posted-content","created":{"date-parts":[[2026,5,23]],"date-time":"2026-05-23T04:55:23Z","timestamp":1779512123000},"source":"Crossref","is-referenced-by-count":0,"title":["Exposure to Antibiotics Modifies the Immune Profiles of Bacterial Extracellular Vesicles from Common Vaginal Anaerobes"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2666-162X","authenticated-orcid":false,"given":"Yu","family":"Hasegawa","sequence":"first","affiliation":[{"name":"Women\u2019s Biomedical Research Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Olivia","family":"Swain","sequence":"additional","affiliation":[{"name":"Women\u2019s Biomedical Research Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Urvija","family":"Rajpal","sequence":"additional","affiliation":[{"name":"Department of Immunology and Immunotherapy, Icahn School of Medicine at Mount Sinai, New York, NY 10029 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"France","sequence":"additional","affiliation":[{"name":"Center for Advanced Microbiome Research and Innovation (CAMRI, Institute for Genome Sciences, and Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore MD 21201 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liqhwa","family":"Ncube","sequence":"additional","affiliation":[{"name":"Women\u2019s Biomedical Research Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ilaria","family":"Mogno","sequence":"additional","affiliation":[{"name":"Department of Immunology and Immunotherapy, Icahn School of Medicine at Mount Sinai, New York, NY 10029 USA"},{"name":"Department of Genetics and Genomics Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hannah","family":"Zierden","sequence":"additional","affiliation":[{"name":"Department of Chemical & Biomolecular Engineering, University of Maryland, Baltimore MD 21201 USA"},{"name":"Robert E. Fischell Institute for Biomedical Devices, University of Maryland, Baltimore MD 21201 USA"},{"name":"Department of Obstetrics, Gynecology and Reproductive Sciences, University of Maryland School of Medicine, Baltimore MD 21201 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0851-2233","authenticated-orcid":false,"given":"Jacques","family":"Ravel","sequence":"additional","affiliation":[{"name":"Center for Advanced Microbiome Research and Innovation (CAMRI, Institute for Genome Sciences, and Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore MD 21201 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michal A.","family":"Elovitz","sequence":"additional","affiliation":[{"name":"Nuttall Women\u2019s Health, New York, NY USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026052617500974000_2026.05.21.726874v1.1","doi-asserted-by":"crossref","unstructured":"Mirzaei, R. et al. 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