{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T21:39:08Z","timestamp":1791581948367,"version":"4.3.4"},"reference-count":45,"publisher":"Public Library of Science (PLoS)","issue":"5","license":[{"start":{"date-parts":[[2024,5,20]],"date-time":"2024-05-20T00:00:00Z","timestamp":1716163200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p><jats:italic>Candida albicans<\/jats:italic> (<jats:italic>C<\/jats:italic>. <jats:italic>albicans<\/jats:italic>) can behave as a commensal yeast colonizing the vaginal mucosa, and in this condition is tolerated by the epithelium. When the epithelial tolerance breaks down, due to <jats:italic>C<\/jats:italic>. <jats:italic>albicans<\/jats:italic> overgrowth and hyphae formation, the generated inflammatory response and cell damage lead to vulvovaginal candidiasis (VVC) symptoms. Here, we focused on the induction of mitochondrial reactive oxygen species (mtROS) in vaginal epithelial cells after <jats:italic>C<\/jats:italic>. <jats:italic>albicans<\/jats:italic> infection and the involvement of fungal burden, morphogenesis and candidalysin (CL) production in such induction. Bioluminescent (BLI) <jats:italic>C<\/jats:italic>. <jats:italic>albicans<\/jats:italic>, <jats:italic>C<\/jats:italic>. <jats:italic>albicans<\/jats:italic> PCA-2 and <jats:italic>C<\/jats:italic>. <jats:italic>albicans<\/jats:italic> 529L strains were employed in an <jats:italic>in vitro<\/jats:italic> infection model including reconstituted vaginal epithelium cells (RVE), produced starting from A-431 cell line. The production of mtROS was kinetically measured by using MitoSOX<jats:sup>\u2122<\/jats:sup> Red probe. The potency of <jats:italic>C<\/jats:italic>. <jats:italic>albicans<\/jats:italic> to induced cell damage to RVE and <jats:italic>C<\/jats:italic>. <jats:italic>albicans<\/jats:italic> proliferation have also been evaluated. <jats:italic>C<\/jats:italic>. <jats:italic>albicans<\/jats:italic> induces a rapid mtROS release from vaginal epithelial cells, in parallel with an increase of the fungal load and hyphal formation. Under the same experimental conditions, the 529L <jats:italic>C<\/jats:italic>. <jats:italic>albicans<\/jats:italic> strain, known to be defective in CL production, induced a minor mtROS release showing the key role of CL in causing epithelial mithocondrial activation. <jats:italic>C<\/jats:italic>. <jats:italic>albicans<\/jats:italic> PCA-2, unable to form hyphae, induced comparable but slower mtROS production as compared to BLI <jats:italic>C<\/jats:italic>. <jats:italic>albicans<\/jats:italic> yeasts. By reducing mtROS through a ROS scavenger, an increased fungal burden was observed during RVE infection but not in fungal cultures grown on abiotic surface. Collectively, we conclude that CL, more than fungal load and hyphae formation, seems to play a key role in the rapid activation of mtROS by epithelial cells and in the induction of cell-damage and that mtROS are key elements in the vaginal epithelial cells response to <jats:italic>C<\/jats:italic>. <jats:italic>albicans<\/jats:italic>.<\/jats:p>","DOI":"10.1371\/journal.pone.0303449","type":"journal-article","created":{"date-parts":[[2024,5,20]],"date-time":"2024-05-20T17:29:52Z","timestamp":1716226192000},"page":"e0303449","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":14,"title":["Fungal burden, dimorphic transition and candidalysin: Role in Candida albicans-induced vaginal cell damage and mitochondrial activation in vitro"],"prefix":"10.1371","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1369-0955","authenticated-orcid":true,"given":"Luca","family":"Spaggiari","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9845-7399","authenticated-orcid":true,"given":"Andrea","family":"Ardizzoni","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesco","family":"Ricchi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-2452-0501","authenticated-orcid":true,"given":"Natalia","family":"Pedretti","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Caterina Alejandra","family":"Squartini Ramos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gianfranco Bruno","family":"Squartini Ramos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samyr","family":"Kenno","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesco","family":"De Seta","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5299-8197","authenticated-orcid":true,"given":"Eva","family":"Pericolini","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2024,5,20]]},"reference":[{"key":"pone.0303449.ref001","doi-asserted-by":"crossref","DOI":"10.12659\/MSM.939088","article-title":"Editorial: The World Health Organization (WHO) Fungal Priority Pathogens List in Response to Emerging Fungal Pathogens During the COVID-19 Pandemic","volume":"28","author":"DV Parums","year":"2022","journal-title":"Med Sci Monit"},{"key":"pone.0303449.ref002","doi-asserted-by":"crossref","first-page":"692491","DOI":"10.3389\/fmicb.2021.692491","article-title":"It Takes Two to Tango: How a Dysregulation of the Innate Immunity, Coupled With Candida Virulence, Triggers VVC Onset","volume":"12","author":"A Ardizzoni","year":"2021","journal-title":"Front Microbiol"},{"key":"pone.0303449.ref003","doi-asserted-by":"crossref","first-page":"2724","DOI":"10.1128\/IAI.00469-16","article-title":"Candida albicans Pathogenesis: Fitting within the Host-Microbe Damage Response Framework","volume":"84","author":"MA Jabra-Rizk","year":"2016","journal-title":"Infect Immun"},{"key":"pone.0303449.ref004","article-title":"Candida pathogens induce protective mitochondria-associated type I interferon signalling and a damage-driven response in vaginal epithelial cells","author":"M Pekmezovic","year":"2021","journal-title":"Nat Microbiol"},{"key":"pone.0303449.ref005","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1038\/nri2975","article-title":"Mitochondria in innate immune responses","volume":"11","author":"AP West","year":"2011","journal-title":"Nat Rev Immunol"},{"key":"pone.0303449.ref006","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1152\/physrev.00044.2005","article-title":"The NOX Family of ROS-Generating NADPH Oxidases: Physiology and Pathophysiology","volume":"87","author":"K Bedard","year":"2007","journal-title":"Physiological Reviews"},{"key":"pone.0303449.ref007","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1016\/j.freeradbiomed.2019.09.029","article-title":"NADPH oxidase in the vasculature: Expression, regulation and signalling pathways; 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physiological signalling agents","volume":"21","author":"H Sies","year":"2020","journal-title":"Nat Rev Mol Cell Biol"},{"key":"pone.0303449.ref025","doi-asserted-by":"crossref","first-page":"foz081","DOI":"10.1093\/femsyr\/foz081","article-title":"Candida albicans infection disturbs the redox homeostasis system and induces reactive oxygen species accumulation for epithelial cell death","volume":"20","author":"T Ren","year":"2020","journal-title":"FEMS Yeast Research"},{"key":"pone.0303449.ref026","doi-asserted-by":"crossref","first-page":"102829","DOI":"10.1016\/j.jbc.2022.102829","article-title":"Candidalysin: Connecting the pore forming mechanism of this virulence factor to its immunostimulatory properties","volume":"299","author":"CM Russell","year":"2023","journal-title":"Journal of Biological Chemistry"},{"key":"pone.0303449.ref027","doi-asserted-by":"crossref","first-page":"e00724","DOI":"10.1128\/mBio.00724-15","article-title":"Secretory Aspartyl Proteinases Cause Vaginitis and Can Mediate Vaginitis Caused by Candida albicans in Mice","volume":"6","author":"E Pericolini","year":"2015","journal-title":"mBio"},{"key":"pone.0303449.ref028","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1038\/nature17625","article-title":"Candidalysin is a fungal peptide toxin critical for mucosal infection","volume":"532","author":"DL Moyes","year":"2016","journal-title":"Nature"},{"key":"pone.0303449.ref029","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/j.mib.2019.06.002","article-title":"Candidalysin: discovery and function in Candida albicans infections","volume":"52","author":"JR Naglik","year":"2019","journal-title":"Current Opinion in Microbiology"},{"key":"pone.0303449.ref030","doi-asserted-by":"crossref","DOI":"10.1128\/mBio.02178-17","article-title":"Processing of Candida albicans Ece1p Is Critical for Candidalysin Maturation and Fungal Virulence","volume":"9","author":"JP Richardson","year":"2018","journal-title":"mBio"},{"key":"pone.0303449.ref031","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1007\/BF02630962","article-title":"Dynamics of differentiation in human epidermoid squamous carcinoma cells (A431) with continuous, long-term gamma-IFN treatment","volume":"27A","author":"J Ridge","year":"1991","journal-title":"In Vitro Cell Dev Biol"},{"key":"pone.0303449.ref032","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1080\/21505594.2016.1213937","article-title":"Therapeutic activity of a Saccharomyces cerevisiae-based probiotic and inactivated whole yeast on vaginal candidiasis","volume":"8","author":"E Pericolini","year":"2017","journal-title":"Virulence"},{"key":"pone.0303449.ref033","doi-asserted-by":"crossref","first-page":"e1005644","DOI":"10.1371\/journal.ppat.1005644","article-title":"Neutrophil Attack Triggers Extracellular Trap-Dependent Candida Cell Wall Remodeling and Altered Immune Recognition","volume":"12","author":"A Hopke","year":"2016","journal-title":"PLoS Pathog"},{"key":"pone.0303449.ref034","doi-asserted-by":"crossref","first-page":"668","DOI":"10.1128\/iai.51.2.668-674.1986","article-title":"Evidence for macrophage-mediated protection against lethal Candida albicans infection","volume":"51","author":"F Bistoni","year":"1986","journal-title":"Infect Immun"},{"key":"pone.0303449.ref035","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1016\/j.micinf.2007.01.012","article-title":"Murine model of concurrent oral and vaginal Candida albicans colonization to study epithelial host\u2013pathogen interactions","volume":"9","author":"D Rahman","year":"2007","journal-title":"Microbes and Infection"},{"key":"pone.0303449.ref036","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1093\/carcin\/bgh079","article-title":"Ascorbic acid suppresses drug-induced apoptosis in human colon cancer cells by scavenging mitochondrial superoxide anions","volume":"25","author":"U Wenzel","year":"2003","journal-title":"Carcinogenesis"},{"key":"pone.0303449.ref037","doi-asserted-by":"crossref","first-page":"e0201436","DOI":"10.1371\/journal.pone.0201436","article-title":"Epitope unmasking in vulvovaginal candidiasis is associated with hyphal growth and neutrophilic infiltration","volume":"13","author":"E Pericolini","year":"2018","journal-title":"PLoS ONE"},{"key":"pone.0303449.ref038","doi-asserted-by":"crossref","first-page":"e26580","DOI":"10.1371\/journal.pone.0026580","article-title":"Candida albicans Yeast and Hyphae are Discriminated by MAPK Signaling in Vaginal Epithelial Cells","volume":"6","author":"DL Moyes","year":"2011","journal-title":"PLoS ONE"},{"key":"pone.0303449.ref039","doi-asserted-by":"crossref","first-page":"1645","DOI":"10.1093\/infdis\/jiz365","article-title":"Vaginal Epithelial Cells Discriminate Between Yeast and Hyphae of Candida albicans in Women Who Are Colonized or Have Vaginal Candidiasis","volume":"220","author":"E Roselletti","year":"2019","journal-title":"J Infect Dis"},{"key":"pone.0303449.ref040","doi-asserted-by":"crossref","first-page":"629","DOI":"10.3389\/fimmu.2017.00629","article-title":"Candida albicans Yeast, Pseudohyphal, and Hyphal Morphogenesis Differentially Affects Immune Recognition","volume":"8","author":"L Mukaremera","year":"2017","journal-title":"Front Immunol"},{"key":"pone.0303449.ref041","doi-asserted-by":"crossref","first-page":"e1009884","DOI":"10.1371\/journal.ppat.1009884","article-title":"A variant ECE1 allele contributes to reduced pathogenicity of Candida albicans during vulvovaginal candidiasis","volume":"17","author":"J Liu","year":"2021","journal-title":"PLoS Pathog"},{"key":"pone.0303449.ref042","doi-asserted-by":"crossref","first-page":"4847","DOI":"10.1128\/IAI.00223-09","article-title":"A multifunctional, synthetic Gaussia princeps luciferase reporter for live imaging of Candida albicans infections","volume":"77","author":"B Enjalbert","year":"2009","journal-title":"Infect Immun"},{"key":"pone.0303449.ref043","doi-asserted-by":"crossref","first-page":"e2213","DOI":"10.21769\/BioProtoc.2213","article-title":"In vitro Detection of Neutrophil Traps and Post-attack Cell Wall Changes in Candida Hyphae","volume":"7","author":"A Hopke","year":"2017","journal-title":"Bio Protoc"},{"key":"pone.0303449.ref044","doi-asserted-by":"crossref","first-page":"40955","DOI":"10.1074\/jbc.M106878200","article-title":"Mechanism of vitamin C inhibition of cell death induced by oxidative stress in glutathione-depleted HL-60 cells","volume":"276","author":"VH Guaiquil","year":"2001","journal-title":"J Biol Chem"},{"key":"pone.0303449.ref045","volume-title":"The GraphPad guide to comparing dose-response or kinetic curves","author":"H Motulsky","year":"1998"}],"container-title":["PLOS 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