{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,22]],"date-time":"2026-06-22T23:50:55Z","timestamp":1782172255768,"version":"3.54.5"},"reference-count":38,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2021,9,29]],"date-time":"2021-09-29T00:00:00Z","timestamp":1632873600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["U54-HD090259"],"award-info":[{"award-number":["U54-HD090259"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["1U01AI124316-01"],"award-info":[{"award-number":["1U01AI124316-01"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["1KL2TR001444"],"award-info":[{"award-number":["1KL2TR001444"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["1R01AI145310-01A1"],"award-info":[{"award-number":["1R01AI145310-01A1"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Methicillin-resistant Staphylococcus aureus (MRSA) has evolved numerous antimicrobial resistance mechanisms and is identified as a serious public health threat by the World Health Organization and U.S. Centers for Disease Control and Prevention. The glycopeptide vancomycin (VAN) remains a cornerstone of therapy for severe MRSA infections despite increasing reports of therapeutic failure in hospitalized patients with bacteremia or pneumonia. Recently, the role of released bacterial-derived membrane vesicles (MVs) in antibiotic resistance has garnered attention. Here we examined the effect of exogenous MRSA-derived MVs on VAN activity against MRSA in vitro, using minimum inhibitory concentration and checkerboard assays, and ex vivo, incorporating components of host innate immunity such as neutrophils and serum complement present in blood. Additionally, the proteome of MVs from VAN-exposed MRSA was characterized to determine if protein expression was altered. The presence of MVs increased the VAN MIC against MRSA to values where clinical failure is commonly observed. Furthermore, the presence of MVs increased survival of MRSA pre-treated with sub-MIC concentrations of VAN in whole blood and upon exposure to human neutrophils but not human serum. Unbiased proteomic analysis also showed an elevated expression of MV proteins associated with antibiotic resistance (e.g., marR) or proteins that are functionally linked to cell membrane\/wall metabolism. Together, our findings indicate MRSA-derived MVs are capable of lowering susceptibility of the pathogen to VAN, whole-blood- and neutrophil-mediated killing, a new pharmacodynamic consideration for a drug increasingly linked to clinical treatment failures.<\/jats:p>","DOI":"10.3390\/microorganisms9102055","type":"journal-article","created":{"date-parts":[[2021,9,30]],"date-time":"2021-09-30T00:03:42Z","timestamp":1632960222000},"page":"2055","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Bacterial Membrane-Derived Vesicles Attenuate Vancomycin Activity against Methicillin-Resistant Staphylococcus aureus"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3867-4268","authenticated-orcid":false,"given":"Monika","family":"Kumaraswamy","sequence":"first","affiliation":[{"name":"Infectious Diseases Section, VA San Diego Healthcare System, San Diego, CA 92161, USA"},{"name":"Department of Medicine, University of California San Diego, La Jolla, CA 92093, USA"},{"name":"Collaborative to Halt Antibiotic Resistant Microbes (CHARM), University of California San Diego, La Jolla, CA 92093, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kamilla","family":"Wiull","sequence":"additional","affiliation":[{"name":"Faculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, 1433 \u00c5s, Norway"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3841-0796","authenticated-orcid":false,"given":"Bishnu","family":"Joshi","sequence":"additional","affiliation":[{"name":"Research Group for Host-Microbe Interactions, UiT-The Arctic University of Norway, 9037 Troms\u00f8, Norway"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"George","family":"Sakoulas","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, University of California San Diego, La Jolla, CA 92093, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Armin","family":"Kousha","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, University of California San Diego, La Jolla, CA 92093, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3077-8003","authenticated-orcid":false,"given":"Gustav","family":"Vaaje-Kolstad","sequence":"additional","affiliation":[{"name":"Faculty of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, 1433 \u00c5s, Norway"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mona","family":"Johannessen","sequence":"additional","affiliation":[{"name":"Research Group for Host-Microbe Interactions, UiT-The Arctic University of Norway, 9037 Troms\u00f8, Norway"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1314-0497","authenticated-orcid":false,"given":"Kristin","family":"Hegstad","sequence":"additional","affiliation":[{"name":"Research Group for Host-Microbe Interactions, UiT-The Arctic University of Norway, 9037 Troms\u00f8, Norway"},{"name":"Norwegian National Advisory Unit on Detection of Antimicrobial Resistance, University Hospital of North-Norway, 9038 Troms\u00f8, Norway"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Victor","family":"Nizet","sequence":"additional","affiliation":[{"name":"Collaborative to Halt Antibiotic Resistant Microbes (CHARM), University of California San Diego, La Jolla, CA 92093, USA"},{"name":"Department of Pediatrics, University of California San Diego, La Jolla, CA 92093, USA"},{"name":"Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, La Jolla, CA 92093, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fatemeh","family":"Askarian","sequence":"additional","affiliation":[{"name":"Research Group for Host-Microbe Interactions, UiT-The Arctic University of Norway, 9037 Troms\u00f8, Norway"},{"name":"Department of Pediatrics, University of California San Diego, La Jolla, CA 92093, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,9,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1001\/archinternmed.2008.536","article-title":"Exclusive Staphylococcus aureus throat carriage: At-risk populations","volume":"169","author":"Mertz","year":"2009","journal-title":"Arch. Intern. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"507","DOI":"10.3389\/fimmu.2013.00507","article-title":"Staphylococcus aureus Colonization: Modulation of Host Immune Response and Impact on Human Vaccine Design","volume":"4","author":"Brown","year":"2014","journal-title":"Front. Immunol."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Tam, K., and Torres, V.J. (2019). Staphylococcus aureus Secreted Toxins and Extracellular Enzymes. Microbiol. Spectr., 7.","DOI":"10.1128\/microbiolspec.GPP3-0039-2018"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"667","DOI":"10.1038\/nrmicro3110","article-title":"Phenol-soluble modulins and staphylococcal infection","volume":"11","author":"Peschel","year":"2013","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"262","DOI":"10.3389\/fmicb.2018.00262","article-title":"Staphylococcus aureus Membrane-Derived Vesicles Promote Bacterial Virulence and Confer Protective Immunity in Murine Infection Models","volume":"9","author":"Askarian","year":"2018","journal-title":"Front. Microbiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1038\/s41467-017-00492-w","article-title":"Prophage-triggered membrane vesicle formation through peptidoglycan damage in Bacillus subtilis","volume":"8","author":"Toyofuku","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1038\/s41579-018-0112-2","article-title":"Types and origins of bacterial membrane vesicles","volume":"17","author":"Toyofuku","year":"2019","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"620","DOI":"10.1038\/nrmicro3480","article-title":"Through the wall: Extracellular vesicles in Gram-positive bacteria, mycobacteria and fungi","volume":"13","author":"Brown","year":"2015","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1948","DOI":"10.1128\/IAI.06014-11","article-title":"Membrane vesicle release in bacteria, eukaryotes, and archaea: A conserved yet underappreciated aspect of microbial life","volume":"80","author":"Deatherage","year":"2012","journal-title":"Infect. Immun."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"514","DOI":"10.1038\/aps.2017.82","article-title":"Versatile effects of bacterium-released membrane vesicles on mammalian cells and infectious\/inflammatory diseases","volume":"39","author":"Yu","year":"2018","journal-title":"Acta Pharmacol. Sin."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"432","DOI":"10.3389\/fmicb.2020.00432","article-title":"Bacterial Membrane Vesicles as Mediators of Microbe\u2014Microbe and Microbe\u2014Host Community Interactions","volume":"11","author":"Caruana","year":"2020","journal-title":"Front. Microbiol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"e01439-18","DOI":"10.1128\/AAC.01439-18","article-title":"Antibiotics Stimulate Formation of Vesicles in Staphylococcus aureus in both Phage-Dependent and -Independent Fashions and via Different Routes","volume":"63","author":"Andreoni","year":"2019","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"16194","DOI":"10.1038\/nmicrobiol.2016.194","article-title":"Staphylococcus aureus inactivates daptomycin by releasing membrane phospholipids","volume":"2","author":"Pader","year":"2016","journal-title":"Nat. Microbiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"21066","DOI":"10.1038\/s41598-020-78121-8","article-title":"Significant increase in the secretion of extracellular vesicles and antibiotics resistance from methicillin-resistant Staphylococcus aureus induced by ampicillin stress","volume":"10","author":"Kim","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_15","unstructured":"CDC (2019). Antibiotic Resistance Threats in the United States."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1146\/annurev.mi.38.100184.002011","article-title":"The structure and mode of action of glycopeptide antibiotics of the vancomycin group","volume":"38","author":"Barna","year":"1984","journal-title":"Annu. Rev. Microbiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2386","DOI":"10.1093\/jac\/dkr301","article-title":"Site of infection rather than vancomycin MIC predicts vancomycin treatment failure in methicillin-resistant Staphylococcus aureus bacteraemia","volume":"66","author":"Walraven","year":"2011","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1264","DOI":"10.1093\/jac\/dkv487","article-title":"Standard susceptibility testing overlooks potent azithromycin activity and cationic peptide synergy against MDR Stenotrophomonas maltophilia","volume":"71","author":"Kumaraswamy","year":"2016","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e00559-16","DOI":"10.1128\/IAI.00559-16","article-title":"Serine-aspartate-repeat protein D Increases Staphylococcus aureus Virulence and Survival in Blood","volume":"85","author":"Askarian","year":"2017","journal-title":"Infect. Immun."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"468","DOI":"10.1016\/j.ijantimicag.2017.12.032","article-title":"Listeria monocytogenes endocarditis: Case report, review of the literature, and laboratory evaluation of potential novel antibiotic synergies","volume":"51","author":"Kumaraswamy","year":"2018","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1016\/j.jprot.2018.05.017","article-title":"Enterococcus faecium produces membrane vesicles containing virulence factors and antimicrobial resistance related proteins","volume":"187","author":"Wagner","year":"2018","journal-title":"J. Proteom."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1016\/j.jprot.2018.11.013","article-title":"Comparative exoproteome profiling of an invasive and a commensal Staphylococcus haemolyticus isolate","volume":"197","author":"Cavanagh","year":"2018","journal-title":"J. Proteom."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"976","DOI":"10.1016\/1044-0305(94)80016-2","article-title":"An approach to correlate tandem mass spectral data of peptides with amino acid sequences in a protein database","volume":"5","author":"Eng","year":"1994","journal-title":"J. Am. Soc. Mass Spectrom."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"D607","DOI":"10.1093\/nar\/gky1131","article-title":"String V11: Protein-Protein Association Networks with Increased Coverage, Supporting Functional Discovery in Genome-Wide Experimental Datasets","volume":"47","author":"Szklarczyk","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Bibek, G.C., Sahukhal, G.S., and Elasri, M.O. (2019). Role of the msaABCR Operon in Cell Wall Biosynthesis, Autolysis, Integrity, and Antibiotic Resistance in Staphylococcus aureus. Antimicrob. Agents Chemother., 63.","DOI":"10.1128\/AAC.00680-19"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2719","DOI":"10.1099\/mic.0.26426-0","article-title":"Genome-wide transcriptional profiling of the response of Staphylococcus aureus to cell-wall-active antibiotics reveals a cell-wall-stress stimulon","volume":"149","author":"Utaida","year":"2003","journal-title":"Microbiology"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1093\/jac\/41.6.581","article-title":"The targeting of factors necessary for expression of methicillin resistance in staphylococci","volume":"41","author":"Labischinski","year":"1998","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"4566","DOI":"10.1128\/AAC.48.12.4566-4573.2004","article-title":"Penicillin-binding protein 2 is essential for expression of high-level vancomycin resistance and cell wall synthesis in vancomycin-resistant Staphylococcus aureus carrying the enterococcal vanA gene complex","volume":"48","author":"Severin","year":"2004","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1815","DOI":"10.1128\/JB.187.5.1815-1824.2005","article-title":"Role of penicillin-binding protein 2 (PBP2) in the antibiotic susceptibility and cell wall cross-linking of Staphylococcus aureus: Evidence for the cooperative functioning of PBP2, PBP4, and PBP2A","volume":"187","author":"Leski","year":"2005","journal-title":"J. Bacteriol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"267","DOI":"10.3947\/ic.2016.48.4.267","article-title":"Treatment of Methicillin-Resistant Staphylococcus aureus Bacteremia","volume":"48","author":"Choo","year":"2016","journal-title":"Infect. Chemother."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2398","DOI":"10.1128\/JCM.42.6.2398-2402.2004","article-title":"Relationship of MIC and bactericidal activity to efficacy of vancomycin for treatment of methicillin-resistant Staphylococcus aureus bacteremia","volume":"42","author":"Sakoulas","year":"2004","journal-title":"J. Clin. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Song, K.H., Kim, M., Kim, C.J., Cho, J.E., Choi, Y.J., Park, J.S., Ahn, S., Jang, H.C., Park, K.H., and Jung, S.I. (2017). Impact of Vancomycin MIC on Treatment Outcomes in Invasive Staphylococcus aureus Infections. Antimicrob. Agents Chemother., 61.","DOI":"10.1128\/AAC.01845-16"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"3315","DOI":"10.1128\/AAC.00113-08","article-title":"Relationship between vancomycin MIC and failure among patients with methicillin-resistant Staphylococcus aureus bacteremia treated with vancomycin","volume":"52","author":"Lodise","year":"2008","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"7868","DOI":"10.1038\/s41598-018-26277-9","article-title":"Risk factors of treatment failure and 30-day mortality in patients with bacteremia due to MRSA with reduced vancomycin susceptibility","volume":"8","author":"Yang","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2215","DOI":"10.1038\/s41598-018-20661-1","article-title":"Genome-scale analysis of Methicillin-resistant Staphylococcus aureus USA300 reveals a tradeoff between pathogenesis and drug resistance","volume":"8","author":"Choe","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"4605","DOI":"10.1128\/JB.00144-09","article-title":"Redundant hydrogen peroxide scavengers contribute to Salmonella virulence and oxidative stress resistance","volume":"191","author":"Hebrard","year":"2009","journal-title":"J. Bacteriol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1093\/jac\/dkm416","article-title":"Does antibiotic exposure increase the risk of methicillin-resistant Staphylococcus aureus (MRSA) isolation? A systematic review and meta-analysis","volume":"61","author":"Tacconelli","year":"2008","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"591","DOI":"10.1038\/nchembio820","article-title":"An oxidation-sensing mechanism is used by the global regulator MgrA in Staphylococcus aureus","volume":"2","author":"Chen","year":"2006","journal-title":"Nat. Chem. Biol."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/9\/10\/2055\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:07:01Z","timestamp":1760166421000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/9\/10\/2055"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,9,29]]},"references-count":38,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2021,10]]}},"alternative-id":["microorganisms9102055"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms9102055","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,9,29]]}}}