{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T17:46:27Z","timestamp":1792604787120,"version":"4.5.0"},"reference-count":55,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,8,23]],"date-time":"2023-08-23T00:00:00Z","timestamp":1692748800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000897","name":"Cystic Fibrosis Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000897","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:p>Lung infection with the fungus <jats:italic>Aspergillus fumigatus (<\/jats:italic>Af<jats:italic>)<\/jats:italic> is a common complication in cystic fibrosis (CF) and is associated with loss of pulmonary function. We established a fungal epithelial co-culture model to examine the impact of Af infection on CF bronchial epithelial barrier function using Af strains 10AF and AF293-GFP, and the CFBE41o- cell line homozygous for the F508del mutation with (CF<jats:sup>+CFTR<\/jats:sup>) and without (CF) normal CFTR expression. Following exposure of the epithelial surface to Af conidia, formation of germlings (early stages of fungal growth) was detected after 9-12 hours and hyphae (mature fungal growth) after 12-24 hours. During fungal morphogenesis, bronchial epithelial cells showed signs of damage including rounding, and partial detachment after 24 hours. Fluorescently labeled conidia were internalized after 6 hours and more internalized conidia were observed in CF compared to CF<jats:sup>+CFTR<\/jats:sup> cells. Infection of the apical surface with 10AF conidia, germlings, or hyphae was performed to determine growth stage-specific effects on tight junction protein zona occludens protein 1 (ZO-1) expression and transepithelial electrical resistance (TER). In response to infection with conidia or germlings, epithelial barrier function degraded time-dependently (based on ZO-1 immunofluorescence and TER) with a delayed onset in CF<jats:sup>+CFTR<\/jats:sup> cell monolayers and required viable fungi and apical application. Infection with hyphae caused an earlier onset and faster rate of decline in TER compared to conidia and germlings. Gliotoxin, a major Af virulence factor, caused a rapid decline in TER and induced a transient chloride secretory response in CF<jats:sup>+CFTR<\/jats:sup> but not CF cells. Our findings suggest growth and internalization of Af result in deleterious effects on bronchial epithelial barrier function that occurred more rapidly in the absence of CFTR. Bronchial epithelial barrier breakdown was time-dependent and morphotype-specific and mimicked by acute administration of gliotoxin. Our study also suggests a protective role for CFTR by turning on CFTR-dependent chloride transport in response to gliotoxin, a mechanism that will support mucociliary clearance, and could delay the loss of epithelial integrity during fungal development <jats:italic>in vivo<\/jats:italic>.<\/jats:p>","DOI":"10.3389\/fcimb.2023.1196581","type":"journal-article","created":{"date-parts":[[2023,8,23]],"date-time":"2023-08-23T19:31:02Z","timestamp":1692819062000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":10,"title":["Protective role of CFTR during fungal infection of cystic fibrosis bronchial epithelial cells with Aspergillus fumigatus"],"prefix":"10.3389","volume":"13","author":[{"given":"Beate","family":"Illek","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Horst","family":"Fischer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Terry E.","family":"Machen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gopika","family":"Hari","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karl V.","family":"Clemons","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriele","family":"Sass","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jose A. G.","family":"Ferreira","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David A.","family":"Stevens","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,8,23]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"126","DOI":"10.1152\/physiologyonline.2001.16.3.126","article-title":"Molecular structure of tight junctions and their role in epithelial transport","volume":"16","author":"Anderson","year":"2001","journal-title":"News Physiol. Sci."},{"key":"B2","doi-asserted-by":"publisher","first-page":"835","DOI":"10.1016\/S0954-6111(99)90270-6","article-title":"Prophylactic antibiotic therapy is associated with an increased prevalence of Aspergillus colonization in adult cystic fibrosis patients","volume":"93","author":"Bargon","year":"1999","journal-title":"Respir. Med."},{"key":"B3","doi-asserted-by":"publisher","first-page":"406","DOI":"10.1111\/j.1365-2141.2010.08283.x","article-title":"Enemy of the (immunosuppressed) state: an update on the pathogenesis of Aspergillus fumigatus infection","volume":"150","author":"Ben-Ami","year":"2010","journal-title":"Br. J. Haematol."},{"key":"B4","doi-asserted-by":"publisher","first-page":"655","DOI":"10.1007\/s11046-021-00541-5","article-title":"CF fungal disease in the age of CFTR modulators","volume":"186","author":"Bercusson","year":"2021","journal-title":"Mycopathologia"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2020.01041","article-title":"Bronchial epithelial cells on the front line to fight lung infection-causing Aspergillus fumigatus","volume":"11","author":"Bigot","year":"2020","journal-title":"Front. Immunol."},{"key":"B6","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1007\/7651_2013_62","article-title":"Multifunction co-culture model for evaluating cell-cell interactions","volume":"1202","author":"Bogdanowicz","year":"2014","journal-title":"Methods Mol. Biol."},{"key":"B7","doi-asserted-by":"publisher","first-page":"1095","DOI":"10.14670\/HH-17.1095","article-title":"Aspergillus fumigatus causes in vitro electrophysiological and morphological modifications in human nasal epithelial cells","volume":"17","author":"Botterel","year":"2002","journal-title":"Histol. Histopathol."},{"key":"B8","doi-asserted-by":"publisher","first-page":"1203","DOI":"10.1136\/thx.51.12.1203","article-title":"Binding of Aspergillus fumigatus spores to lung epithelial cells and basement membrane proteins: relevance to the asthmatic lung","volume":"51","author":"Bromley","year":"1996","journal-title":"Thorax"},{"key":"B9","doi-asserted-by":"publisher","first-page":"379","DOI":"10.1111\/2049-632X.12152","article-title":"Increased production of gliotoxin is related to the formation of biofilm by Aspergillus fumigatus: an immunological approach","volume":"70","author":"Bugli","year":"2014","journal-title":"Pathog. Dis."},{"key":"B10","doi-asserted-by":"publisher","first-page":"229","DOI":"10.2147\/IDR.S63621","article-title":"Aspergillus fumigatus in the cystic fibrosis lung: pros and cons of azole therapy","volume":"9","author":"Burgel","year":"2016","journal-title":"Infect. Drug Resist."},{"key":"B11","doi-asserted-by":"publisher","first-page":"301","DOI":"10.1164\/rccm.201106-1027OC","article-title":"Cystic fibrosis transmembrane conductance regulator regulates epithelial cell response to Aspergillus and resultant pulmonary inflammation","volume":"185","author":"Chaudhary","year":"2012","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"B12","doi-asserted-by":"publisher","first-page":"1084","DOI":"10.1016\/j.jcf.2022.09.007","article-title":"Advent of elexacaftor\/tezacaftor\/ivacaftor for cystic fibrosis treatment: What consequences on Aspergillus-related diseases? Preliminary insights","volume":"21","author":"Chesnay","year":"2022","journal-title":"J. Cyst Fibros"},{"key":"B13","doi-asserted-by":"publisher","first-page":"723129","DOI":"10.1155\/2013\/723129","article-title":"Aspergillus-associated airway disease, inflammation, and the innate immune response","volume":"2013","author":"Chotirmall","year":"2013","journal-title":"BioMed. Res. Int."},{"key":"B14","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1111\/2049-632X.12180","article-title":"Mimicking the host and its microenvironment in vitro for studying mucosal infections by Pseudomonas aeruginosa","volume":"71","author":"Crabbe","year":"2014","journal-title":"Pathog. Dis."},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2016.00472","article-title":"Interactions of Aspergillus fumigatus conidia with airway epithelial cells: A critical review","volume":"7","author":"Croft","year":"2016","journal-title":"Front. Microbiol."},{"key":"B16","doi-asserted-by":"publisher","first-page":"447","DOI":"10.1128\/CMR.00055-08","article-title":"Pathogenesis of Aspergillus fumigatus in invasive aspergillosis","volume":"22","author":"Dagenais","year":"2009","journal-title":"Clin. Microbiol. Rev."},{"key":"B17","doi-asserted-by":"publisher","first-page":"146","DOI":"10.1093\/infdis\/175.1.146","article-title":"Binding and germination of Aspergillus fumigatus conidia on cultured A549 pneumocytes","volume":"175","author":"DeHart","year":"1997","journal-title":"J. Infect. Dis."},{"key":"B18","doi-asserted-by":"publisher","first-page":"1151","DOI":"10.1093\/infdis\/162.5.1151","article-title":"Restriction endonuclease analysis of total cellular DNA of Aspergillus fumigatus isolates of geographically and epidemiologically diverse origin","volume":"162","author":"Denning","year":"1990","journal-title":"J. Infect. Dis."},{"key":"B19","doi-asserted-by":"publisher","first-page":"852419","DOI":"10.1155\/2011\/852419","article-title":"Epithelial cell coculture models for studying infectious diseases: benefits and limitations","volume":"2011","author":"Duell","year":"2011","journal-title":"J. BioMed. Biotechnol."},{"key":"B20","doi-asserted-by":"publisher","first-page":"2172264","DOI":"10.1080\/21505594.2023.2172264","article-title":"Pathogenicity and virulence of Aspergillus fumigatus","volume":"14","author":"Earle","year":"2023","journal-title":"Virulence"},{"key":"B21","doi-asserted-by":"publisher","first-page":"477","DOI":"10.1038\/nature04712","article-title":"The ABC protein turned chloride channel whose failure causes cystic fibrosis","volume":"440","author":"Gadsby","year":"2006","journal-title":"Nature"},{"key":"B22","doi-asserted-by":"publisher","first-page":"e24997","DOI":"10.4161\/tisb.24997","article-title":"Barrier function of airway tract epithelium","volume":"1","author":"Ganesan","year":"2013","journal-title":"Tissue Barriers"},{"key":"B23","doi-asserted-by":"publisher","first-page":"3717","DOI":"10.4049\/jimmunol.176.6.3717","article-title":"Dectin-1 and TLRs permit macrophages to distinguish between different Aspergillus fumigatus cellular states","volume":"176","author":"Gersuk","year":"2006","journal-title":"J. Immunol."},{"key":"B24","doi-asserted-by":"publisher","first-page":"1953","DOI":"10.1128\/EC.00274-07","article-title":"Aspergillus fumigatus: principles of pathogenesis and host defense","volume":"6","author":"Hohl","year":"2007","journal-title":"Eukaryot Cell"},{"key":"B25","doi-asserted-by":"publisher","first-page":"442","DOI":"10.1016\/j.jmii.2022.12.004","article-title":"COVID-19 associated mold infections: Review of COVID-19 associated pulmonary aspergillosis and mucormycosis","volume":"56","author":"Huang","year":"2022","journal-title":"J. Microbiol. Immunol. Infect"},{"key":"B26","doi-asserted-by":"publisher","first-page":"57","DOI":"10.1159\/000149783","article-title":"Cl transport in complemented CF bronchial epithelial cells correlates with CFTR mRNA expression levels","volume":"22","author":"Illek","year":"2008","journal-title":"Cell Physiol. Biochem."},{"key":"B27","doi-asserted-by":"publisher","first-page":"983","DOI":"10.1159\/000324011","article-title":"Sensitivity of chloride efflux vs. transepithelial measurements in mixed CF and normal airway epithelial cell populations","volume":"26","author":"Illek","year":"2010","journal-title":"Cell Physiol. Biochem."},{"key":"B28","doi-asserted-by":"publisher","first-page":"S115","DOI":"10.1080\/13693780600897989","article-title":"Interactions of Aspergillus fumigatus with vascular endothelial cells","volume":"44","author":"Kamai","year":"2006","journal-title":"Med. Mycol."},{"key":"B29","doi-asserted-by":"publisher","first-page":"S95","DOI":"10.1080\/13693780500051547","article-title":"Aspergillus mycotoxins and their effect on the host","author":"Kamei","year":"2005","journal-title":"Med. Mycol."},{"key":"B30","doi-asserted-by":"publisher","first-page":"5","DOI":"10.1186\/1471-2180-7-5","article-title":"Verruculogen associated with Aspergillus fumigatus hyphae and conidia modifies the electrophysiological properties of human nasal epithelial cells","volume":"7","author":"Khoufache","year":"2007","journal-title":"BMC Microbiol."},{"key":"B31","doi-asserted-by":"publisher","first-page":"a009589","DOI":"10.1101\/cshperspect.a009589","article-title":"CFTR, mucins, and mucus obstruction in cystic fibrosis","volume":"2","author":"Kreda","year":"2012","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"B32","doi-asserted-by":"publisher","first-page":"591","DOI":"10.1080\/13693780600815411","article-title":"Gliotoxin-mediated suppression of innate and adaptive immune functions directed against Listeria monocytogenes","volume":"44","author":"Kupfahl","year":"2006","journal-title":"Med. Mycol."},{"key":"B33","doi-asserted-by":"publisher","first-page":"S97","DOI":"10.1016\/j.tim.2009.08.004","article-title":"What do we know about the role of gliotoxin in the pathobiology of Aspergillus fumigatus","volume":"47","author":"Kwon-Chung","year":"2009","journal-title":"Med. Mycol."},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00140-18","article-title":"Aspergillus fumigatus and aspergillosis in 2019","author":"Latge","year":"2019","journal-title":"Clin. Microbiol. Rev"},{"key":"B35","doi-asserted-by":"publisher","first-page":"635","DOI":"10.1128\/IAI.73.1.635-637.2005","article-title":"Detection of gliotoxin in experimental and human aspergillosis","volume":"73","author":"Lewis","year":"2005","journal-title":"Infect. Immun."},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2018.02514","article-title":"A co-culture model of PBMC and stem cell derived human nasal epithelium reveals rapid activation of NK and innate T cells upon influenza A virus infection of the nasal epithelium","volume":"9","author":"Luukkainen","year":"2018","journal-title":"Front. Immunol."},{"key":"B37","doi-asserted-by":"publisher","first-page":"3279","DOI":"10.1099\/00221287-146-12-3279","article-title":"Landmarks in the early duplication cycles of Aspergillus fumigatus and Aspergillus nidulans: polarity, germ tube emergence and septation","author":"Momany","year":"2000","journal-title":"Microbiol. (Reading)"},{"key":"B38","doi-asserted-by":"publisher","first-page":"525","DOI":"10.1111\/j.1365-2222.1978.tb01506.x","article-title":"Fungal spores in lung and sputum","volume":"8","author":"Mullins","year":"1978","journal-title":"Clin. Allergy"},{"key":"B39","doi-asserted-by":"publisher","first-page":"1151","DOI":"10.1038\/nature04332","article-title":"Genomic sequence of the pathogenic and allergenic filamentous fungus Aspergillus fumigatus","volume":"438","author":"Nierman","year":"2005","journal-title":"Nature"},{"key":"B40","first-page":"Abstract no. 24","article-title":"Mycotoxin secretion by Aspergillus fumigatus as a response to mycovirus infection","author":"Patil","year":"2022"},{"key":"B41","doi-asserted-by":"publisher","first-page":"1824","DOI":"10.1086\/323900","article-title":"The impact of culture isolation of Aspergillus species: a hospital-based survey of aspergillosis","volume":"33","author":"Perfect","year":"2001","journal-title":"Clin. Infect. Dis."},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2018.01205","article-title":"Aspergillus fumigatus inhibits Pseudomonas aeruginosa in Co-culture: Implications of a mutually antagonistic relationship on virulence and inflammation in the CF airway","volume":"9","author":"Reece","year":"2018","journal-title":"Front. Microbiol."},{"key":"B43","doi-asserted-by":"publisher","first-page":"1653","DOI":"10.1016\/j.freeradbiomed.2008.09.011","article-title":"Oxidative stress caused by pyocyanin impairs CFTR Cl- transport in human bronchial epithelial cells","volume":"45","author":"Schwarzer","year":"2008","journal-title":"Free Radic. Biol. Med."},{"key":"B44","doi-asserted-by":"publisher","first-page":"34850","DOI":"10.1074\/jbc.M110.167668","article-title":"Pseudomonas aeruginosa Homoserine lactone activates store-operated cAMP and cystic fibrosis transmembrane regulator-dependent Cl- secretion by human airway epithelia","volume":"285","author":"Schwarzer","year":"2010","journal-title":"J. Biol. Chem."},{"key":"B45","doi-asserted-by":"publisher","first-page":"G1129","DOI":"10.1152\/ajpgi.2000.279.6.G1129","article-title":"Molecular physiology and pathophysiology of tight junctions V. Assault of the tight junction by enteric pathogens","volume":"279","author":"Sears","year":"2000","journal-title":"Am. J. Physiol. Gastrointest Liver Physiol."},{"key":"B46","doi-asserted-by":"publisher","first-page":"1870","DOI":"10.1056\/NEJMra0808853","article-title":"Aspergillosis","volume":"360","author":"Segal","year":"2009","journal-title":"N Engl. J. Med."},{"key":"B47","doi-asserted-by":"publisher","first-page":"375","DOI":"10.1016\/j.mib.2011.07.006","article-title":"Molecular mechanism of Aspergillus fumigatus adherence to host constituents","volume":"14","author":"Sheppard","year":"2011","journal-title":"Curr. Opin. Microbiol."},{"key":"B48","doi-asserted-by":"publisher","first-page":"2814","DOI":"10.1128\/JB.01567-07","article-title":"The type III toxins of Pseudomonas aeruginosa disrupt epithelial barrier function","volume":"190","author":"Soong","year":"2008","journal-title":"J. Bacteriol"},{"key":"B49","doi-asserted-by":"publisher","first-page":"107","DOI":"10.1177\/2211068214561025","article-title":"TEER measurement techniques for in vitro barrier model systems","volume":"20","author":"Srinivasan","year":"2015","journal-title":"J. Lab. Autom"},{"key":"B50","doi-asserted-by":"publisher","first-page":"349","DOI":"10.1309\/UJRV9DLC11RM3G8R","article-title":"Host-dependent patterns of tissue injury in invasive pulmonary aspergillosis","volume":"127","author":"Stergiopoulou","year":"2007","journal-title":"Am. J. Clin. Pathol."},{"key":"B51","doi-asserted-by":"publisher","first-page":"123","DOI":"10.1016\/j.jcf.2007.06.006","article-title":"Prevalence and antimicrobial susceptibility of microorganisms isolated from sputa of patients with cystic fibrosis","volume":"7","author":"Valenza","year":"2008","journal-title":"J. Cyst Fibros"},{"key":"B52","doi-asserted-by":"publisher","first-page":"S118","DOI":"10.1093\/mmy\/myy074","article-title":"Aspergillus colonization and antifungal immunity in cystic fibrosis patients","volume":"57","author":"Warris","year":"2019","journal-title":"Med. Mycol"},{"key":"B53","doi-asserted-by":"publisher","first-page":"3377","DOI":"10.1128\/IAI.68.6.3377-3384.2000","article-title":"Adhesion of Aspergillus species to extracellular matrix proteins: evidence for involvement of negatively charged carbohydrates on the conidial surface","volume":"68","author":"Wasylnka","year":"2000","journal-title":"Infect. Immun."},{"key":"B54","doi-asserted-by":"publisher","first-page":"F31","DOI":"10.1083\/jcb.151.5.F31","article-title":"Tight junction, a platform for trafficking and signaling protein complexes","volume":"151","author":"Zahraoui","year":"2000","journal-title":"J. Cell Biol."},{"key":"B55","doi-asserted-by":"publisher","first-page":"C68","DOI":"10.1152\/ajpcell.00043.2017","article-title":"Airway epithelial anion secretion and barrier function following exposure to fungal aeroallergens: role of oxidative stress","volume":"313","author":"Zaidman","year":"2017","journal-title":"Am. J. Physiol. Cell Physiol."}],"container-title":["Frontiers in Cellular and Infection Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2023.1196581\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,8,23]],"date-time":"2023-08-23T19:31:05Z","timestamp":1692819065000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2023.1196581\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,8,23]]},"references-count":55,"alternative-id":["10.3389\/fcimb.2023.1196581"],"URL":"https:\/\/doi.org\/10.3389\/fcimb.2023.1196581","relation":{},"ISSN":["2235-2988"],"issn-type":[{"value":"2235-2988","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,8,23]]},"article-number":"1196581"}}