{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T18:25:40Z","timestamp":1787077540751,"version":"build-2736575974"},"reference-count":37,"publisher":"Public Library of Science (PLoS)","issue":"8","license":[{"start":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T00:00:00Z","timestamp":1787011200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100015691","name":"Division of Microbiology and Infectious Diseases, National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["2R01AI093692"],"award-info":[{"award-number":["2R01AI093692"]}],"id":[{"id":"10.13039\/100015691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>\n                    <jats:italic>Chlamydia trachomatis<\/jats:italic>\n                    infection remains prevalent, with high rates of reinfection. Most infections are asymptomatic, and without treatment, women are at risk for reproductive and perinatal complications that are exacerbated with repeat infections. Currently, no\n                    <jats:italic>C. trachomatis<\/jats:italic>\n                    vaccine is available nor are there clinical biomarkers to predict reinfection risk. Small, non-coding microRNAs (miRNAs) are promising biomarker candidates because of their easy isolation, high stability, and role as molecular messengers. MiRNAs can be intracellular and\/or extracellular, with the latter being released from cells and either packaged into extracellular vesicles (EVs) or freely circulating in association with proteins. We identified miRNAs from serum EVs and whole serum collected from 42 women (53 samples) and 35 women (49 samples), respectively. Sera were collected at a baseline treatment visit and a 3-month follow-up visit, where women were determined to be reinfected or not reinfected by nucleic acid amplification test. MiRNA profiles were evaluated using Bruker nCounter microarrays, data were positive ligation normalized, miRNAs were categorized as detected or not detected, nominal significance was calculated using Fisher\u2019s exact test, and predicted target mRNAs were assessed with Qiagen\u2019s Ingenuity Pathway Analysis (IPA). Detection of EV-derived miR-888-5p at baseline was associated with reinfection at the follow-up visit. For whole serum, detection of miR-1285-5p, \u2212548aa\u2009+\u2009548t-3p, and \u2212575 at baseline were associated with absence of reinfection at follow-up. MiRs-888-5p and \u2212548aa aligned with varying degrees to mRNA targets associated with CD8+ and CD4\u2009+\u2009T-cell functions, with miR-888-5p demonstrating strong CD8\u2009+\u2009T-cell associations while miR-548aa was largely associated with CD4\u2009+\u2009T-cell responses. Distinct EV and whole serum miRNAs were identified as potential biomarker candidates that are associated with reinfection risk. Our findings are preliminary, and future work includes validation studies to confirm whether these miRNAs can predict\n                    <jats:italic>C. trachomatis<\/jats:italic>\n                    reinfection risk.\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0356267","type":"journal-article","created":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T17:35:09Z","timestamp":1787074509000},"page":"e0356267","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Evaluation of extracellular microRNAs as potential biomarker candidates for assessing chlamydia reinfection risk"],"prefix":"10.1371","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2301-9575","authenticated-orcid":true,"given":"Chloe Meewes","family":"Lewis","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kanupriya","family":"Gupta","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Howard","family":"Wiener","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hemant K.","family":"Tiwari","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William M.","family":"Geisler","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,8,18]]},"reference":[{"key":"pone.0356267.ref001","volume-title":"Global health sector strategies on, respectively, HIV, viral hepatitis and sexually transmitted infections for the period 2022\u20132030","author":"World Health Organization","year":"2022"},{"key":"pone.0356267.ref002","doi-asserted-by":"crossref","DOI":"10.1136\/bmj.c1642","article-title":"Randomised controlled trial of screening for Chlamydia trachomatis to prevent pelvic inflammatory disease: the POPI (prevention of pelvic infection) trial","volume":"340","author":"P Oakeshott","year":"2010","journal-title":"BMJ"},{"issue":"5","key":"pone.0356267.ref003","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1136\/sextrans-2019-053999","article-title":"Pregnancy and fertility-related adverse outcomes associated with Chlamydia trachomatis infection: a global systematic review and meta-analysis","volume":"96","author":"W Tang","year":"2020","journal-title":"Sex Transm Infect"},{"issue":"8","key":"pone.0356267.ref004","doi-asserted-by":"crossref","first-page":"478","DOI":"10.1097\/OLQ.0b013e3181a2a933","article-title":"Repeat infection with Chlamydia and gonorrhea among females: a systematic review of the literature","volume":"36","author":"CB Hosenfeld","year":"2009","journal-title":"Sex Transm Dis"},{"key":"pone.0356267.ref005","doi-asserted-by":"crossref","first-page":"1197","DOI":"10.3389\/fmicb.2020.01197","article-title":"MicroRNA Biomarkers for Infectious Diseases: From Basic Research to Biosensing","volume":"11","author":"L Tribolet","year":"2020","journal-title":"Front Microbiol"},{"key":"pone.0356267.ref006","doi-asserted-by":"crossref","first-page":"402","DOI":"10.3389\/fendo.2018.00402","article-title":"Overview of MicroRNA Biogenesis, Mechanisms of Actions, and Circulation","volume":"9","author":"J O\u2019Brien","year":"2018","journal-title":"Front Endocrinol (Lausanne)"},{"issue":"3","key":"pone.0356267.ref007","doi-asserted-by":"crossref","first-page":"483","DOI":"10.1161\/CIRCRESAHA.111.247452","article-title":"Circulating microRNAs: novel biomarkers and extracellular communicators in cardiovascular disease?","volume":"110","author":"EE Creemers","year":"2012","journal-title":"Circ Res"},{"issue":"8","key":"pone.0356267.ref008","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1038\/nrm3838","article-title":"Regulation of microRNA biogenesis","volume":"15","author":"M Ha","year":"2014","journal-title":"Nat Rev Mol Cell Biol"},{"key":"pone.0356267.ref009","doi-asserted-by":"crossref","DOI":"10.3402\/jev.v3.23743","article-title":"Exosomes provide a protective and enriched source of miRNA for biomarker profiling compared to intracellular and cell-free blood","volume":"3","author":"L Cheng","year":"2014","journal-title":"J Extracell Vesicles"},{"key":"pone.0356267.ref010","doi-asserted-by":"crossref","first-page":"1174","DOI":"10.3389\/fmicb.2019.01174","article-title":"Small RNA Profiles of Serum Exosomes Derived From Individuals With Latent and Active Tuberculosis","volume":"10","author":"L Lyu","year":"2019","journal-title":"Front Microbiol"},{"key":"pone.0356267.ref011","doi-asserted-by":"crossref","first-page":"661","DOI":"10.3389\/fimmu.2018.00661","article-title":"A Serum Circulating miRNA Signature for Short-Term Risk of Progression to Active Tuberculosis Among Household Contacts","volume":"9","author":"FJ Duffy","year":"2018","journal-title":"Front Immunol"},{"issue":"8","key":"pone.0356267.ref012","doi-asserted-by":"crossref","first-page":"105397","DOI":"10.1016\/j.micinf.2024.105397","article-title":"Role of microRNAs in immune regulation and pathogenesis of Chlamydia trachomatis and Chlamydia muridarum infections: a rapid review","volume":"26","author":"C Meewes","year":"2024","journal-title":"Microbes Infect"},{"issue":"3","key":"pone.0356267.ref013","doi-asserted-by":"crossref","DOI":"10.1128\/mBio.01241-14","article-title":"Early microRNA expression profile as a prognostic biomarker for the development of pelvic inflammatory disease in a mouse model of chlamydial genital infection","volume":"5","author":"L Yeruva","year":"2014","journal-title":"mBio"},{"issue":"1","key":"pone.0356267.ref014","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1186\/s12864-019-5495-6","article-title":"A unique insight into the MiRNA profile during genital chlamydial infection","volume":"20","author":"I Benyeogor","year":"2019","journal-title":"BMC Genomics"},{"issue":"3","key":"pone.0356267.ref015","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pntd.0002117","article-title":"Conjunctival MicroRNA expression in inflammatory trachomatous scarring","volume":"7","author":"T Derrick","year":"2013","journal-title":"PLoS Negl Trop Dis"},{"key":"pone.0356267.ref016","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1186\/s12879-016-1367-8","article-title":"Inverse relationship between microRNA-155 and -184 expression with increasing conjunctival inflammation during ocular Chlamydia trachomatis infection","volume":"16","author":"T Derrick","year":"2016","journal-title":"BMC Infect Dis"},{"issue":"2","key":"pone.0356267.ref017","doi-asserted-by":"crossref","DOI":"10.1093\/femspd\/ftx016","article-title":"miRNAs that associate with conjunctival inflammation and ocular Chlamydia trachomatis infection do not predict progressive disease","volume":"75","author":"T Derrick","year":"2017","journal-title":"Pathog Dis"},{"issue":"1","key":"pone.0356267.ref018","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1186\/s12866-020-02061-6","article-title":"Host cell response and distinct gene expression profiles at different stages of Chlamydia trachomatis infection reveals stage-specific biomarkers of infection","volume":"21","author":"EE Dzakah","year":"2021","journal-title":"BMC Microbiol"},{"issue":"10","key":"pone.0356267.ref019","doi-asserted-by":"crossref","DOI":"10.1128\/IAI.00057-20","article-title":"Endocervical miRNA Expression Profiles in Women Positive for Chlamydia trachomatis with Clinical Signs and\/or Symptoms Are Distinct from Those in Women Positive for Chlamydia trachomatis without Signs and Symptoms","volume":"88","author":"TA Batteiger","year":"2020","journal-title":"Infect Immun"},{"key":"pone.0356267.ref020","doi-asserted-by":"crossref","first-page":"1981","DOI":"10.3389\/fimmu.2018.01981","article-title":"An Adaptive Chlamydia trachomatis-Specific IFN-\u03b3-Producing CD4+ T Cell Response Is Associated With Protection Against Chlamydia Reinfection in Women","volume":"9","author":"RK Bakshi","year":"2018","journal-title":"Front Immunol"},{"issue":"4","key":"pone.0356267.ref021","doi-asserted-by":"crossref","DOI":"10.1128\/CVI.00010-17","article-title":"The Predominant CD4+ Th1 Cytokine Elicited to Chlamydia trachomatis Infection in Women Is Tumor Necrosis Factor Alpha and Not Interferon Gamma","volume":"24","author":"SJ Jordan","year":"2017","journal-title":"Clin Vaccine Immunol"},{"key":"pone.0356267.ref022","first-page":"1","article-title":"Sexually transmitted disease guidelines, 2015","volume":"64","author":"Centers for Disease Control and Prevention","year":"2015","journal-title":"MMWR Recomm Rep"},{"key":"pone.0356267.ref023","doi-asserted-by":"crossref","first-page":"133","DOI":"10.12688\/f1000research.103533.2","article-title":"Laboratory protocol for the digital multiplexed gene expression analysis of nasopharyngeal swab samples using the NanoString nCounter system","volume":"11","author":"M Garc\u00eda Aranda","year":"2022","journal-title":"F1000Res"},{"key":"pone.0356267.ref024","unstructured":"Bruker Spatial Biology, Inc. nCounter miRNA expression analysis in plasma and serum samples. 2019."},{"issue":"1","key":"pone.0356267.ref025","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1002\/1097-0142(1950)3:1<32::AID-CNCR2820030106>3.0.CO;2-3","article-title":"Index for rating diagnostic tests","volume":"3","author":"WJ Youden","year":"1950","journal-title":"Cancer"},{"issue":"9","key":"pone.0356267.ref026","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0162445","article-title":"Intravaginal Chlamydia trachomatis Challenge Infection Elicits TH1 and TH17 Immune Responses in Mice That Promote Pathogen Clearance and Genital Tract Damage","volume":"11","author":"RD Vicetti Miguel","year":"2016","journal-title":"PLoS One"},{"issue":"5","key":"pone.0356267.ref027","doi-asserted-by":"crossref","DOI":"10.1093\/immhor\/vlaf004","article-title":"Chlamydia trachomatis-specific interferon-\u03b3-producing CD8 T-cells are associated with lower chlamydia bacterial load in reinfected women","volume":"9","author":"WM Geisler","year":"2025","journal-title":"Immunohorizons"},{"issue":"11","key":"pone.0356267.ref028","doi-asserted-by":"crossref","first-page":"3782","DOI":"10.1002\/(SICI)1521-4141(199911)29:11<3782::AID-IMMU3782>3.0.CO;2-B","article-title":"IFN-gamma knockout mice show Th2-associated delayed-type hypersensitivity and the inflammatory cells fail to localize and control chlamydial infection","volume":"29","author":"S Wang","year":"1999","journal-title":"Eur J Immunol"},{"issue":"2","key":"pone.0356267.ref029","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1038\/nri1551","article-title":"Immunology of Chlamydia infection: implications for a Chlamydia trachomatis vaccine","volume":"5","author":"RC Brunham","year":"2005","journal-title":"Nat Rev Immunol"},{"issue":"11","key":"pone.0356267.ref030","doi-asserted-by":"crossref","first-page":"2043","DOI":"10.1093\/infdis\/jiab610","article-title":"Problems With Understanding Chlamydia trachomatis Immunology","volume":"225","author":"RC Brunham","year":"2022","journal-title":"J Infect Dis"},{"issue":"12","key":"pone.0356267.ref031","doi-asserted-by":"crossref","first-page":"4661","DOI":"10.1128\/iai.63.12.4661-4668.1995","article-title":"Gene knockout mice establish a primary protective role for major histocompatibility complex class II-restricted responses in Chlamydia trachomatis genital tract infection","volume":"63","author":"RP Morrison","year":"1995","journal-title":"Infect Immun"},{"issue":"1","key":"pone.0356267.ref032","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1038\/s41541-020-0157-x","article-title":"Parenteral vaccination protects against transcervical infection with Chlamydia trachomatis and generate tissue-resident T cells post-challenge","volume":"5","author":"NDNT Nguyen","year":"2020","journal-title":"NPJ Vaccines"},{"issue":"10","key":"pone.0356267.ref033","doi-asserted-by":"crossref","first-page":"4648","DOI":"10.4049\/jimmunol.1400120","article-title":"CD8+ T cells define an unexpected role in live-attenuated vaccine protective immunity against Chlamydia trachomatis infection in macaques","volume":"192","author":"N Olivares-Zavaleta","year":"2014","journal-title":"J Immunol"},{"key":"pone.0356267.ref034","doi-asserted-by":"crossref","first-page":"1679688","DOI":"10.3389\/fimmu.2026.1679688","article-title":"miRNA\/mRNA analysis of increased TGF-\u03b2 pathways drive epithelial-mesenchymal transition and regulatory T cell differentiation","volume":"17","author":"T Darville","year":"2026","journal-title":"Front Immunol"},{"issue":"18","key":"pone.0356267.ref035","doi-asserted-by":"crossref","first-page":"6486","DOI":"10.3390\/ijms21186486","article-title":"Extracellular Vesicles: New Endogenous Shuttles for miRNAs in Cancer Diagnosis and Therapy?","volume":"21","author":"S Martellucci","year":"2020","journal-title":"Int J Mol Sci"},{"key":"pone.0356267.ref036","doi-asserted-by":"crossref","first-page":"1951","DOI":"10.1016\/j.csbj.2024.04.061","article-title":"Challenges and opportunities in processing NanoString nCounter data","volume":"23","author":"J Chilimoniuk","year":"2024","journal-title":"Comput Struct Biotechnol J"},{"issue":"6","key":"pone.0356267.ref037","doi-asserted-by":"crossref","first-page":"801","DOI":"10.1016\/j.jmoldx.2020.03.005","article-title":"Comparison of GeneChip, nCounter, and Real-Time PCR-Based Gene Expressions Predicting Locoregional Tumor Control after Primary and Postoperative Radiochemotherapy in Head and Neck Squamous Cell Carcinoma","volume":"22","author":"S Schmidt","year":"2020","journal-title":"J Mol Diagn"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0356267","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T17:35:15Z","timestamp":1787074515000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0356267"}},"subtitle":[],"editor":[{"given":"David M.","family":"Ojcius","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,8,18]]},"references-count":37,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,8,18]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0356267","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,18]]}}}