{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T12:28:26Z","timestamp":1790166506719,"version":"4.1.0"},"reference-count":111,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2026,3,7]],"date-time":"2026-03-07T00:00:00Z","timestamp":1772841600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100014055","name":"USAMRAA","doi-asserted-by":"publisher","award":["HT94252410629"],"award-info":[{"award-number":["HT94252410629"]}],"id":[{"id":"10.13039\/100014055","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100013759","name":"Commonwealth Health Research Board","doi-asserted-by":"publisher","award":["#247-09-24"],"award-info":[{"award-number":["#247-09-24"]}],"id":[{"id":"10.13039\/100013759","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100019828","name":"Virginia Innovation Partnership Corporation","doi-asserted-by":"crossref","award":["CCF23-0104-HE"],"award-info":[{"award-number":["CCF23-0104-HE"]}],"id":[{"id":"10.13039\/100019828","id-type":"DOI","asserted-by":"crossref"}]},{"name":"HRSA\/LymeX Diagnostics Prize\/Phase1","award":["206271"],"award-info":[{"award-number":["206271"]}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>The cause of chronic neurological effects associated with Lyme disease (LD) remains unclear. We propose that bacterial extracellular vesicles (BEVs) released by Borrelia burgdorferi, the causative agent of LD, exacerbate spirochete-induced damage and serve as a persistent source of antigenic stimulation. We showed that, over a 10-day period, in vitro cultures of B. burgdorferi B31 produced 38,000 BEVs per spirochete with a distinctive double-membrane structure and median diameter of 143.3 nm. BEVs contained known immunogenic and immunomodulatory molecules such as peptidoglycan, p66, flagellar filament protein (FlaB), basic membrane proteins A\/B\/D, BdrV, GroEL, CRASP-1, ErpA8, glycerophosphodiester phosphodiesterase, p37, OMS28, p13, OspA\/B\/C, VlsE, and outer membrane glycolipids (e.g., cholesteryl 6-O acyl beta D galactopyranoside). Chromosome-encoded 16S ribosomal RNA and cp32 plasmid-encoded OspE and terminase genes were also detected in the BEVs. Of the 45 Borrelia proteins identified in the urine of a C3H\/HeJ murine model of Lyme disease, 14 were associated with BEVs. In human urine samples, 31 of 289 spirochete proteins detected in patients with either acute Lyme disease or persistent borreliosis post-treatment symptoms, including p66 and FlaB, were also BEV-associated. BEV treatment of HMC3 human microglial cells reduced phagocytic activity and triggered aberrant activation of inflammatory and immunometabolic pathways, including upregulation of interferon-alpha (IFN-\u03b1), aconitate decarboxylase 1 (Acod1), and Toll-like receptor 2 (TLR2) gene expression. BEVs also induced NRF2 nuclear translocation. In conclusion, these findings support that BEVs can amplify spirochete-induced damage and act as antigenic debris, driving dampened phagocytic activity and dysregulated inflammation, with implications for diagnostics and therapeutics targeting vesicle-mediated pathology.<\/jats:p>","DOI":"10.3390\/microorganisms14030600","type":"journal-article","created":{"date-parts":[[2026,3,9]],"date-time":"2026-03-09T11:06:52Z","timestamp":1773054412000},"page":"600","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Characterization\u00a0of\u00a0Borrelia-Derived Extracellular Vesicles: Implications for Pathogenesis and Diagnostics"],"prefix":"10.3390","volume":"14","author":[{"given":"Barbara","family":"Birkaya","sequence":"first","affiliation":[{"name":"Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA 20110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ahana","family":"Byne","sequence":"additional","affiliation":[{"name":"Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA 20110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sumaiya","family":"Irfan","sequence":"additional","affiliation":[{"name":"Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA 20110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph","family":"Gallagher","sequence":"additional","affiliation":[{"name":"Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA 20110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dominic","family":"Granato","sequence":"additional","affiliation":[{"name":"Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA 20110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hayat","family":"Kharmoud","sequence":"additional","affiliation":[{"name":"Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA 20110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-3943-0403","authenticated-orcid":false,"given":"Andrea Blake","family":"Brothers","sequence":"additional","affiliation":[{"name":"College Dean\u2019s Office, American University, Washington, DC 20016, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1008-4271","authenticated-orcid":false,"given":"Elsa","family":"Ronzier","sequence":"additional","affiliation":[{"name":"Biomedical Research Laboratory, Institute for Biohealth Innovation, George Mason University, 10650 Pyramid Place, Manassas, VA 20110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amanda Haymond","family":"Still","sequence":"additional","affiliation":[{"name":"Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA 20110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8802-3597","authenticated-orcid":false,"given":"Weidong","family":"Zhou","sequence":"additional","affiliation":[{"name":"Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA 20110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5016-8694","authenticated-orcid":false,"given":"Robert K.","family":"Ernst","sequence":"additional","affiliation":[{"name":"Department of Microbial Pathogenesis, University of Maryland School of Dentistry, Baltimore, MD 21021, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hope","family":"McIntyre","sequence":"additional","affiliation":[{"name":"Lyme Hope, LLC, Mount Airy, MD 21771, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6674-7532","authenticated-orcid":false,"given":"Ashley Michelle","family":"Groshong","sequence":"additional","affiliation":[{"name":"Laboratory of Bacteriology, Rocky Mountain Laboratories, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT 59840, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5155-7907","authenticated-orcid":false,"given":"Lance A.","family":"Liotta","sequence":"additional","affiliation":[{"name":"Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA 20110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1599-0214","authenticated-orcid":false,"given":"Alessandra","family":"Luchini","sequence":"additional","affiliation":[{"name":"Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA 20110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,3,7]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"102536","DOI":"10.1016\/j.ttbdis.2025.102536","article-title":"Proposing a subgenus Borreliella","volume":"16","author":"Margos","year":"2025","journal-title":"Ticks Tick-Borne Dis."},{"key":"ref_2","unstructured":"CDC (2025, June 23). Lyme Disease Surveillance and Data, Available online: https:\/\/www.cdc.gov\/lyme\/data-research\/facts-stats\/index.html."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1418","DOI":"10.1212\/CON.0000000000001334","article-title":"Infectious Neuropathies","volume":"29","author":"Boegle","year":"2023","journal-title":"Continuum"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Govil, S., Capitle, E., Lacqua, A., Khianey, R., Coyle, P., and Schutzer, S.E. (2023). Common Neurologic Features of Lyme Disease That May Present to a Rheumatologist. Pathogens, 12.","DOI":"10.3390\/pathogens12040576"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Fallon, B.A., Kuvaldina, M., Zubcevik, N., DeBiasi, R., Mulkey, S.B., Chiu, C., Chow, F., Paolino, K., Lai, R., and Putrino, D. (2025). Proposed research classification criteria for Lyme disease in infection associated chronic illness studies. Front. Med., 12.","DOI":"10.3389\/fmed.2025.1519163"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e443","DOI":"10.1016\/j.ijid.2013.01.008","article-title":"Development of a foundation for a case definition of post-treatment Lyme disease syndrome","volume":"17","author":"Aucott","year":"2013","journal-title":"Int. J. Infect. Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1007\/s11136-012-0126-6","article-title":"Post-treatment Lyme disease syndrome symptomatology and the impact on life functioning: Is there something here?","volume":"22","author":"Aucott","year":"2013","journal-title":"Qual. Life Res."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Rebman, A.W., and Aucott, J.N. (2020). Post-treatment Lyme Disease as a Model for Persistent Symptoms in Lyme Disease. Front. Med., 7.","DOI":"10.3389\/fmed.2020.00057"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e040399","DOI":"10.1136\/bmjopen-2020-040399","article-title":"Symptom heterogeneity and patient subgroup classification among US patients with post-treatment Lyme disease: An observational study","volume":"11","author":"Rebman","year":"2021","journal-title":"BMJ Open"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"13498","DOI":"10.1073\/pnas.1904170116","article-title":"Borrelia burgdorferi peptidoglycan is a persistent antigen in patients with Lyme arthritis","volume":"116","author":"Jutras","year":"2019","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"McCausland, J.W., Kloos, Z.A., Irnov, I., Sonnert, N.D., Zhou, J., Putnick, R., Mueller, E.A., Steere, A.C., Palm, N.W., and Grimes, C.L. (2025). Bacterial and host enzymes modulate the inflammatory response produced by the peptidoglycan of the Lyme disease agent. bioRxiv.","DOI":"10.1101\/2025.01.08.631998"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"672","DOI":"10.1016\/j.ajpath.2017.11.005","article-title":"Late Disseminated Lyme Disease: Associated Pathology and Spirochete Persistence Posttreatment in Rhesus Macaques","volume":"188","author":"Crossland","year":"2018","journal-title":"Am. J. Pathol."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Gadila, S.K.G., Rosoklija, G., Dwork, A.J., Fallon, B.A., and Embers, M.E. (2021). Detecting Borrelia Spirochetes: A Case Study with Validation Among Autopsy Specimens. Front. Neurol., 12.","DOI":"10.3389\/fneur.2021.628045"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e0007422","DOI":"10.1128\/cmr.00074-22","article-title":"Persistent Borrelia burgdorferi","volume":"35","author":"Verschoor","year":"2022","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"666554","DOI":"10.3389\/fmed.2021.666554","article-title":"Recent Progress in Lyme Disease and Remaining Challenges","volume":"8","author":"Bobe","year":"2021","journal-title":"Front. Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2652","DOI":"10.1172\/JCI58813","article-title":"Spirochete antigens persist near cartilage after murine Lyme borreliosis therapy","volume":"122","author":"Bockenstedt","year":"2012","journal-title":"J. Clin. Investig."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"110651","DOI":"10.1016\/j.celrep.2022.110651","article-title":"Uptake, functionality, and re-release of extracellular vesicle-encapsulated cargo","volume":"39","author":"Ughetto","year":"2022","journal-title":"Cell Rep."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"22131","DOI":"10.1038\/s41598-022-26217-8","article-title":"Fibroblast growth factor-2 bound to specific dermal fibroblast-derived extracellular vesicles is protected from degradation","volume":"12","author":"Petit","year":"2022","journal-title":"Sci. Rep."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e12159","DOI":"10.1002\/jev2.12159","article-title":"Visualizing transfer of microbial biomolecules by outer membrane vesicles in microbe-host-communication in vivo","volume":"10","author":"Bittel","year":"2021","journal-title":"J. Extracell. Vesicles"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1038\/s41392-022-01056-1","article-title":"Pseudomonas aeruginosa: Pathogenesis, virulence factors, antibiotic resistance, interaction with host, technology advances and emerging therapeutics","volume":"7","author":"Qin","year":"2022","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1096","DOI":"10.1016\/j.tim.2024.04.003","article-title":"Staphylococcus aureus membrane vesicles: An evolving story","volume":"32","author":"Wang","year":"2024","journal-title":"Trends Microbiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1415","DOI":"10.1111\/j.1365-2958.2009.06613.x","article-title":"Comparative cryo-electron tomography of pathogenic Lyme disease spirochetes","volume":"71","author":"Kudryashev","year":"2009","journal-title":"Mol. Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Karvonen, K., Tammisto, H., Nykky, J., and Gilbert, L. (2022). Borrelia burgdorferi Outer Membrane Vesicles Contain Antigenic Proteins, but Do Not Induce Cell Death in Human Cells. Microorganisms, 10.","DOI":"10.3390\/microorganisms10020212"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"3652","DOI":"10.1172\/JCI39401","article-title":"Live imaging reveals a biphasic mode of dissemination of Borrelia burgdorferi within ticks","volume":"119","author":"Caimano","year":"2009","journal-title":"J. Clin. Investig."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2265015","DOI":"10.1080\/21505594.2023.2265015","article-title":"Pathogenicity and virulence of Borrelia burgdorferi","volume":"14","author":"Strnad","year":"2023","journal-title":"Virulence"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"e12093","DOI":"10.1002\/jev2.12093","article-title":"Urinary extracellular vesicles: A position paper by the Urine Task Force of the International Society for Extracellular Vesicles","volume":"10","author":"Blijdorp","year":"2021","journal-title":"J. Extracell Vesicles"},{"key":"ref_27","first-page":"11040","article-title":"Urine: The Original Liquid Biopsy","volume":"29","author":"Gomela","year":"2022","journal-title":"Can J. Urol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1089\/gtmb.2018.0189","article-title":"Urine-Based Liquid Biopsy for Nonurological Cancers","volume":"23","author":"Jain","year":"2019","journal-title":"Genet. Test. Mol. Biomarkers"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"112538","DOI":"10.1016\/j.isci.2025.112538","article-title":"Lactobacillus rhamnosus-derived extracellular vesicles influence calcium deposition in a model of breast cancer intraductal calcium stress","volume":"28","author":"Vuong","year":"2025","journal-title":"iScience"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1186\/s41232-022-00224-8","article-title":"Helicobacter pylori infection in the stomach induces neuroinflammation: The potential roles of bacterial outer membrane vesicles in an animal model of Alzheimer\u2019s disease","volume":"42","author":"Park","year":"2022","journal-title":"Inflamm. Regen."},{"key":"ref_31","first-page":"227","article-title":"Do Bacterial Outer Membrane Vesicles Contribute to Chronic Inflammation in Parkinson\u2019s Disease?","volume":"14","author":"Koukoulis","year":"2024","journal-title":"J. Park. Dis."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2159","DOI":"10.1016\/j.cell.2025.02.007","article-title":"Peripheral nervous system microglia-like cells regulate neuronal soma size throughout evolution","volume":"188","author":"Wu","year":"2025","journal-title":"Cell"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Simpson, D.S.A., and Oliver, P.L. (2020). ROS Generation in Microglia: Understanding Oxidative Stress and Inflammation in Neurodegenerative Disease. Antioxidants, 9.","DOI":"10.3390\/antiox9080743"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2026","DOI":"10.1080\/15548627.2023.2167689","article-title":"Inhibition of autophagy in microglia and macrophages exacerbates innate immune responses and worsens brain injury outcomes","volume":"19","author":"Hegdekar","year":"2023","journal-title":"Autophagy"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e12306","DOI":"10.1002\/jev2.12306","article-title":"Helicobacter pylori-derived outer membrane vesicles contribute to Alzheimer\u2019s disease pathogenesis via C3-C3aR signalling","volume":"12","author":"Xie","year":"2023","journal-title":"J. Extracell. Vesicles"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Sun, B., Sawant, H., Borthakur, A., and Bihl, J.C. (2023). Emerging therapeutic role of gut microbial extracellular vesicles in neurological disorders. Front. Neurosci., 17.","DOI":"10.3389\/fnins.2023.1241418"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"e93","DOI":"10.1002\/jex2.93","article-title":"The use of a multicellular in vitro model to investigate uptake and migration of bacterial extracellular vesicles derived from the human gut commensal Bacteroides thetaiotaomicron","volume":"2","author":"Modasia","year":"2023","journal-title":"J Extracell Biol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Ha, J.Y., Choi, S.-Y., Lee, J.H., Hong, S.-H., and Lee, H.-J. (2020). Delivery of Periodontopathogenic Extracellular Vesicles to Brain Monocytes and Microglial IL-6 Promotion by RNA Cargo. Front. Mol. Biosci., 7.","DOI":"10.3389\/fmolb.2020.596366"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Petnicki-Ocwieja, T., McCarthy, J.E., Powale, U., Langston, P.K., Helble, J.D., and Hu, L.T. (2023). Borrelia burgdorferi initiates early transcriptional re-programming in macrophages that supports long-term suppression of inflammation. PLoS Pathog., 19.","DOI":"10.1371\/journal.ppat.1011886"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"822","DOI":"10.1038\/s41423-020-0489-5","article-title":"ACOD1 in immunometabolism and disease","volume":"17","author":"Wu","year":"2020","journal-title":"Cell. Mol. Immunol."},{"key":"ref_41","first-page":"56482","article-title":"Sample Preparation and Imaging of Exosomes by Transmission Electron Microscopy","volume":"131","author":"Jung","year":"2018","journal-title":"J. Vis. Exp."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"3737","DOI":"10.1038\/s41596-021-00566-6","article-title":"Tutorial: Best practices and considerations for mass-spectrometry-based protein biomarker discovery and validation","volume":"16","author":"Nakayasu","year":"2021","journal-title":"Nat. Protoc."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"e345","DOI":"10.1016\/S1473-3099(24)00211-1","article-title":"Plasma-based antigen persistence in the post-acute phase of COVID-19","volume":"24","author":"Peluso","year":"2024","journal-title":"Lancet Infect Dis."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Casselli, T., Divan, A., Vomhof-DeKrey, E.E., Tourand, Y., Pecoraro, H.L., and Brissette, C.A. (2021). A murine model of Lyme disease demonstrates that Borrelia burgdorferi colonizes the dura mater and induces inflammation in the central nervous system. PLoS Pathog., 17.","DOI":"10.1371\/journal.ppat.1009256"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"e00442-19","DOI":"10.1128\/IAI.00442-19","article-title":"Generality of Post-Antimicrobial Treatment Persistence of Borrelia burgdorferi Strains N40 and B31 in Genetically Susceptible and Resistant Mouse Strains","volume":"87","author":"Hodzic","year":"2019","journal-title":"Infect. Immun."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"3186","DOI":"10.1128\/IAI.02036-14","article-title":"Identification of lysine residues in the Borrelia burgdorferi DbpA Adhesin required for murine infection","volume":"82","author":"Fortune","year":"2014","journal-title":"Infect. Immun."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"3693","DOI":"10.1128\/IAI.00828-15","article-title":"BB0744 Affects Tissue Tropism and Spatial Distribution of Borrelia burgdorferi","volume":"83","author":"Wager","year":"2015","journal-title":"Infect. Immun."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1016\/j.nano.2017.11.020","article-title":"Toward detection of toxoplasmosis from urine in mice using hydro-gel nanoparticles concentration and parallel reaction monitoring mass spectrometry","volume":"14","author":"Steinberg","year":"2018","journal-title":"Nanomed. Nanotechnol. Biol. Med."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"19340","DOI":"10.1038\/s41598-020-75051-3","article-title":"Evaluation of pathogen specific urinary peptides in tick-borne illnesses","volume":"10","author":"Magni","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Steinberg, H.E., Bowman, N.M., Diestra, A., Ferradas, C., Russo, P., Clark, D.E., Zhu, D., Magni, R., Malaga, E., and Diaz, M. (2021). Detection of toxoplasmic encephalitis in HIV positive patients in urine with hydrogel nanoparticles. PLoS Neglected Trop. Dis., 15.","DOI":"10.1371\/journal.pntd.0009199"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"19178","DOI":"10.1021\/ja207515j","article-title":"Multifunctional core\u2013shell nanoparticles: Discovery of previously invisible biomarkers","volume":"133","author":"Tamburro","year":"2011","journal-title":"J. Am. Chem. Soc."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"502","DOI":"10.1007\/s12274-008-8054-z","article-title":"Concentration and Preservation of very Low Abundance Biomarkers in Urine, Such as Human Growth Hormone (hGH), by Cibacron Blue F3G-A Loaded Hydrogel Particles","volume":"1","author":"Fredolini","year":"2008","journal-title":"Nano Res."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"353","DOI":"10.1080\/14789450.2018.1450631","article-title":"Affinity enrichment for mass spectrometry: Improving the yield of low abundance biomarkers","volume":"15","author":"Kim","year":"2018","journal-title":"Expert Rev. Proteom."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1007\/978-1-0716-3561-2_9","article-title":"Identification of Unambiguous Borrelia Peptides in Human Urine Using Affinity Capture and Mass Spectrometry","volume":"2742","author":"Cornero","year":"2024","journal-title":"Methods Mol. Biol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"7913","DOI":"10.1073\/pnas.1232451100","article-title":"A newly discovered cholesteryl galactoside from Borrelia burgdorferi","volume":"100","author":"Coxon","year":"2003","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"14","DOI":"10.33696\/immunology.7.218","article-title":"Urinary Borrelia Peptides Correlate with the General Symptom Questionnaire (GSQ-30) Scores in Symptomatic Patients with Suspicion of Tick-borne Illness","volume":"7","author":"Datla","year":"2025","journal-title":"J. Cell. Immunol."},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"Gonz\u00e1lez-Carnicero, Z., Hernanz, R., Mart\u00ednez-Casales, M., Barr\u00fas, M.T., Mart\u00edn, \u00c1., and Alonso, M.J. (2023). Regulation by Nrf2 of IL-1\u03b2-induced inflammatory and oxidative response in VSMC and its relationship with TLR4. Front. Pharmacol., 14.","DOI":"10.3389\/fphar.2023.1058488"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"6215","DOI":"10.1007\/s10072-022-06280-9","article-title":"Infectious diseases and cognition: Do we have to worry?","volume":"43","author":"Letenneur","year":"2022","journal-title":"Neurol. Sci."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"678","DOI":"10.1002\/acr2.11710","article-title":"HLA-DR-Expressing Fibroblast-Like Synoviocytes Are Inducible Antigen Presenting Cells That Present Autoantigens in Lyme Arthritis","volume":"6","author":"Rouse","year":"2024","journal-title":"ACR Open Rheumatol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"S62","DOI":"10.1093\/infdis\/jiae203","article-title":"Persistent Symptoms After Lyme Disease: Clinical Characteristics, Predictors, and Classification","volume":"230","author":"Baarsma","year":"2024","journal-title":"J. Infect. Dis."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"6951","DOI":"10.1038\/s41388-020-01509-3","article-title":"Emerging role of bacterial extracellular vesicles in cancer","volume":"39","author":"Chronopoulos","year":"2020","journal-title":"Oncogene"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"e12085","DOI":"10.1002\/jev2.12085","article-title":"Biodistribution of extracellular vesicles following administration into animals: A systematic review","volume":"10","author":"Kang","year":"2021","journal-title":"J. Extracell. Vesicles"},{"key":"ref_63","doi-asserted-by":"crossref","unstructured":"Cooke, A.C., Nello, A.V., Ernst, R.K., and Schertzer, J.W. (2019). Analysis of Pseudomonas aeruginosa biofilm membrane vesicles supports multiple mechanisms of biogenesis. PLoS ONE, 14.","DOI":"10.1371\/journal.pone.0212275"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"2330768","DOI":"10.1080\/21645515.2024.2330768","article-title":"Intranasal delivery of Salmonella OMVs decorated with Chlamydia trachomatis antigens induces specific local and systemic immune responses","volume":"20","author":"Huynh","year":"2024","journal-title":"Hum Vaccin Immunother. Hum. Vaccin. Immunother."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"e12514","DOI":"10.1002\/jev2.12514","article-title":"Enhancing protective immunity against bacterial infection via coating nano-Rehmannia glutinosa polysaccharide with outer membrane vesicles","volume":"13","author":"Huang","year":"2024","journal-title":"J. Extracell. Vesicles"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1186\/s12974-023-02728-7","article-title":"Helicobacter pylori outer membrane vesicles induce astrocyte reactivity through nuclear factor-\u03baappa B activation and cause neuronal damage in vivo in a murine model","volume":"20","author":"Palacios","year":"2023","journal-title":"J. Neuroinflammation"},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"e70135","DOI":"10.1002\/iid3.70135","article-title":"Porphyromonas gingivalis and Its Outer Membrane Vesicles Induce Neuroinflammation in Mice Through Distinct Mechanisms","volume":"13","author":"Qiu","year":"2025","journal-title":"Immun. Inflamm. Dis."},{"key":"ref_68","doi-asserted-by":"crossref","unstructured":"Yang, B., Li, X., Fu, C., Cai, W., Meng, B., Qu, Y., Kou, X., and Zhang, Q. (2023). Extracellular vesicles in osteoarthritis of peripheral joint and temporomandibular joint. Front. Endocrinol., 14.","DOI":"10.3389\/fendo.2023.1158744"},{"key":"ref_69","first-page":"57","article-title":"Exosomes in intercellular communication and implications for osteoarthritis","volume":"59","author":"Asghar","year":"2020","journal-title":"Rheumatology"},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1089\/met.2022.0109","article-title":"Bacterial Extracellular Vesicle Composition in Human Urine and the 10-Year Risk of Abdominal Obesity","volume":"21","author":"Shin","year":"2023","journal-title":"Metab. Syndr. Relat. Disord."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"e12521","DOI":"10.1002\/jev2.12521","article-title":"The emerging role of bacterial extracellular vesicles in human cancers","volume":"13","author":"Liang","year":"2024","journal-title":"J. Extracell. Vesicles"},{"key":"ref_72","doi-asserted-by":"crossref","unstructured":"Zhou, W., Woodson, M., Neupane, B., Bai, F., Sherman, M.B., Choi, K.H., Neelakanta, G., and Sultana, H. (2018). Exosomes serve as novel modes of tick-borne flavivirus transmission from arthropod to human cells and facilitates dissemination of viral RNA and proteins to the vertebrate neuronal cells. PLoS Pathog., 14.","DOI":"10.1371\/journal.ppat.1006764"},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1186\/s40168-020-00881-2","article-title":"The extracellular vesicle of gut microbial Paenalcaligenes hominis is a risk factor for vagus nerve-mediated cognitive impairment","volume":"8","author":"Lee","year":"2020","journal-title":"Microbiome"},{"key":"ref_74","doi-asserted-by":"crossref","unstructured":"Jan, A.T. (2017). Outer Membrane Vesicles (OMVs) of Gram-negative Bacteria: A Perspective Update. Front. Microbiol., 8.","DOI":"10.3389\/fmicb.2017.01053"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"102015","DOI":"10.1016\/j.isci.2020.102015","article-title":"Mycolic acid-containing bacteria trigger distinct types of membrane vesicles through different routes","volume":"24","author":"Nagakubo","year":"2021","journal-title":"iScience"},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1080\/09687688.2017.1400602","article-title":"Bacterial outer membrane vesicles: New insights and applications","volume":"33","author":"Anand","year":"2016","journal-title":"Mol. Membr. Biol."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"2645","DOI":"10.1101\/gad.1299905","article-title":"Bacterial outer membrane vesicles and the host\u2013pathogen interaction","volume":"19","author":"Kuehn","year":"2005","journal-title":"Genes Dev."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"4159","DOI":"10.1128\/IAI.69.6.4159-4163.2001","article-title":"Influence of cultivation media on genetic regulatory patterns in Borrelia burgdorferi","volume":"69","author":"Yang","year":"2001","journal-title":"Infect. Immun."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"e108174","DOI":"10.15252\/embj.2021108174","article-title":"The extracellular vesicle generation paradox: A bacterial point of view","volume":"40","author":"McMillan","year":"2021","journal-title":"EMBO J."},{"key":"ref_80","unstructured":"Trevisan, G. (2024). Lyme Borreliosis, Edizioni Minerva Medica."},{"key":"ref_81","doi-asserted-by":"crossref","unstructured":"Crowley, J.T., Toledo, A.M., LaRocca, T.J., Coleman, J.L., London, E., and Benach, J.L. (2013). Lipid exchange between Borrelia burgdorferi and host cells. PLoS Pathog., 9.","DOI":"10.1371\/journal.ppat.1003109"},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"e0025022","DOI":"10.1128\/iai.00250-22","article-title":"Erp and Rev Adhesins of the Lyme Disease Spirochete\u2019s Ubiquitous cp32 Prophages Assist the Bacterium during Vertebrate Infection","volume":"91","author":"Stevenson","year":"2023","journal-title":"Infect. Immun."},{"key":"ref_83","doi-asserted-by":"crossref","unstructured":"Ren, Z., Takacs, C.N., Brand\u00e3o, H.B., Jacobs-Wagner, C., and Wang, X. (2023). Organization and replicon interactions within the highly segmented genome of Borrelia burgdorferi. PLoS Genet., 19.","DOI":"10.1101\/2023.03.19.532819"},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"e51","DOI":"10.1038\/emi.2015.56","article-title":"Persister mechanisms in Borrelia burgdorferi: Implications for improved intervention","volume":"4","author":"Feng","year":"2015","journal-title":"Emerg. Microbes Infect."},{"key":"ref_85","first-page":"109","article-title":"Structural features of Borrelia burgdorferi--the Lyme disease spirochete: Silver staining for nucleic acids","volume":"3","author":"Garon","year":"1989","journal-title":"Scanning Microsc. Suppl."},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"7072","DOI":"10.1038\/s41598-017-07288-4","article-title":"Bacterial membrane vesicles transport their DNA cargo into host cells","volume":"7","author":"Bitto","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"740","DOI":"10.1038\/35047123","article-title":"A Toll-like receptor recognizes bacterial DNA","volume":"408","author":"Hemmi","year":"2000","journal-title":"Nature"},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"786","DOI":"10.1126\/science.1232458","article-title":"Cyclic GMP-AMP Synthase is a cytosolic DNA sensor that activates the type I interferon pathway","volume":"339","author":"Sun","year":"2013","journal-title":"Science"},{"key":"ref_89","doi-asserted-by":"crossref","first-page":"e0517922","DOI":"10.1128\/spectrum.05179-22","article-title":"Planktonic and Biofilm-Derived Pseudomonas aeruginosa Outer Membrane Vesicles Facilitate Horizontal Gene Transfer of Plasmid DNA","volume":"11","author":"Johnston","year":"2023","journal-title":"Microbiol. Spectr."},{"key":"ref_90","doi-asserted-by":"crossref","unstructured":"Fridmanis, J., Bobrovs, R., Brangulis, K., T\u0101rs, K., and Jaudzems, K. (2020). Structural and Functional Analysis of BBA03, Borrelia burgdorferi Competitive Advantage Promoting Outer Surface Lipoprotein. Pathogens, 9.","DOI":"10.3390\/pathogens9100826"},{"key":"ref_91","doi-asserted-by":"crossref","first-page":"484","DOI":"10.1128\/CMR.18.3.484-509.2005","article-title":"Diagnosis of lyme borreliosis","volume":"18","author":"Wang","year":"2005","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_92","doi-asserted-by":"crossref","unstructured":"Cafiero, T., and Toledo, A. (2021). Borrelia burgdorferi Surface Exposed GroEL Is a Multifunctional Protein. Pathogens, 10.","DOI":"10.3390\/pathogens10020226"},{"key":"ref_93","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1128\/IAI.05671-11","article-title":"A surface enolase participates in borrelia burgdorferi-plasminogen interaction and contributes to pathogen survival within feeding ticks","volume":"80","author":"Nogueira","year":"2012","journal-title":"Infect. Immun."},{"key":"ref_94","doi-asserted-by":"crossref","first-page":"490","DOI":"10.1086\/653825","article-title":"Complement regulator\u2013acquiring surface protein 1 of Borrelia burgdorferi binds to human bone morphogenic protein 2, Several extracellular matrix proteins, and plasminogen","volume":"202","author":"Haupt","year":"2010","journal-title":"J. Infect. Dis."},{"key":"ref_95","doi-asserted-by":"crossref","first-page":"471","DOI":"10.1128\/microbiolspec.MDNA3-0038-2014","article-title":"vlsAntigenic Variation Systems of Lyme Disease Borrelia: Eluding Host Immunity through both Random, Segmental Gene Conversion and Framework Heterogeneity","volume":"2","author":"Norris","year":"2014","journal-title":"Microbiol. Spectr."},{"key":"ref_96","doi-asserted-by":"crossref","first-page":"372","DOI":"10.1016\/j.tim.2013.06.005","article-title":"Illuminating the roles of the Borrelia burgdorferi adhesins","volume":"21","author":"Coburn","year":"2013","journal-title":"Trends Microbiol."},{"key":"ref_97","doi-asserted-by":"crossref","first-page":"350","DOI":"10.1016\/j.tim.2007.06.003","article-title":"Hidden in plain sight: Borrelia burgdorferi and the extracellular matrix","volume":"15","author":"Cabello","year":"2007","journal-title":"Trends Microbiol."},{"key":"ref_98","doi-asserted-by":"crossref","first-page":"3138","DOI":"10.1128\/IAI.72.6.3138-3146.2004","article-title":"Borrelia burgdorferi binds to, invades, and colonizes native type I collagen lattices","volume":"72","author":"Zambrano","year":"2004","journal-title":"Infect. Immun."},{"key":"ref_99","doi-asserted-by":"crossref","first-page":"105453","DOI":"10.1016\/j.archoralbio.2022.105453","article-title":"Porphyromonas gingivalis outer membrane vesicles modulate cytokine and chemokine production by gingipain-dependent mechanisms in human macrophages","volume":"140","author":"Castillo","year":"2022","journal-title":"Arch. Oral Biol."},{"key":"ref_100","doi-asserted-by":"crossref","first-page":"eaal2807","DOI":"10.1126\/scitranslmed.aal2807","article-title":"Urine lipoarabinomannan glycan in HIV-negative patients with pulmonary tuberculosis correlates with disease severity","volume":"9","author":"Paris","year":"2017","journal-title":"Sci. Transl. Med."},{"key":"ref_101","first-page":"359","article-title":"Diagnosis of toxoplasmosis reactivation in HIV patients in urine using nanoparticle technology","volume":"95","author":"Steinberg","year":"2017","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_102","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1186\/s12014-024-09462-z","article-title":"Recent progress in mass spectrometry-based urinary proteomics","volume":"21","author":"Joshi","year":"2024","journal-title":"Clin. Proteom."},{"key":"ref_103","doi-asserted-by":"crossref","first-page":"38517","DOI":"10.18632\/oncotarget.15870","article-title":"Identification of urine biomarkers associated with lung adenocarcinoma","volume":"8","author":"Wang","year":"2017","journal-title":"Oncotarget"},{"key":"ref_104","doi-asserted-by":"crossref","unstructured":"Jiang, B., and Huang, J. (2024). Influences of bacterial extracellular vesicles on macrophage immune functions. Front. Cell. Infect. Microbiol., 14.","DOI":"10.3389\/fcimb.2024.1411196"},{"key":"ref_105","doi-asserted-by":"crossref","first-page":"2409","DOI":"10.3201\/eid2911.230839","article-title":"Investigation of Possible Link between Interferon-\u03b1 and Lyme Disease","volume":"29","author":"Alaedini","year":"2023","journal-title":"Emerg. Infect. Dis."},{"key":"ref_106","doi-asserted-by":"crossref","unstructured":"Tal, M.C., Hansen, P.S., Ogasawara, H.A., Feng, Q., Volk, R.F., Lee, B., Casebeer, S.E., Blacker, G.S., Shoham, M., and Galloway, S.D. (2024). P66 is a bacterial mimic of CD47 that binds the anti-phagocytic receptor SIRP\u03b1 and facilitates macrophage evasion by Borrelia burgdorferi. bioRxiv.","DOI":"10.1101\/2024.04.29.591704"},{"key":"ref_107","doi-asserted-by":"crossref","first-page":"4848","DOI":"10.1128\/IAI.01215-15","article-title":"Outer surface protein OspC is an antiphagocytic factor that protects borrelia burgdorferi from phagocytosis by macrophages","volume":"83","author":"Carrasco","year":"2015","journal-title":"Infect. Immun."},{"key":"ref_108","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1177\/10815589241290206","article-title":"Human microglia polarization following infection with the Lyme disease spirochete","volume":"73","author":"Akinlusi","year":"2025","journal-title":"J. Investig. Med."},{"key":"ref_109","doi-asserted-by":"crossref","unstructured":"Myers, T.A., Kaushal, D., and Philipp, M.T. (2009). Microglia are mediators of borrelia burgdorferi\u2013induced apoptosis in SH-SY5Y neuronal cells. PLoS Pathog., 5.","DOI":"10.1371\/journal.ppat.1000659"},{"key":"ref_110","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1186\/1742-2094-9-249","article-title":"Acetate supplementation reduces microglia activation and brain interleukin-1\u03b2 Levels in a rat model of lyme neuroborreliosis","volume":"9","author":"Houdek","year":"2012","journal-title":"J. Neuroinflammation"},{"key":"ref_111","doi-asserted-by":"crossref","unstructured":"Greenmyer, J.R., Gaultney, R.A., Brissette, C.A., and Watt, J.A. (2018). Primary Human Microglia Are Phagocytically Active and Respond to Borrelia burgdorferi with Upregulation of Chemokines and Cytokines. Front. Microbiol., 9.","DOI":"10.3389\/fmicb.2018.00811"}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/3\/600\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,10]],"date-time":"2026-03-10T05:11:12Z","timestamp":1773119472000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/3\/600"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,7]]},"references-count":111,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2026,3]]}},"alternative-id":["microorganisms14030600"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms14030600","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,7]]}}}