{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T04:29:49Z","timestamp":1789187389235,"version":"build-2803163510"},"reference-count":42,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,4,10]],"date-time":"2026-04-10T00:00:00Z","timestamp":1775779200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Air\u2013liquid interface (ALI) cell culture systems have improved the study of host-pathogen interactions in respiratory infections. However, most ALI models lack immune components, limiting their ability to capture epithelial\u2013immune crosstalk. To address this, we developed a dual-cell ALI model incorporating human peripheral blood monocyte-derived macrophages beneath differentiated airway epithelial cells.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methodology<\/jats:title>\n                    <jats:p>\n                      Macrophages were seeded on the basolateral side of transwell inserts using fibronectin coating. Model characterization included transepithelial electrical resistance (TEER) to assess epithelial barrier integrity, IL-8 secretion as a marker of epithelial inflammatory signaling, and confocal microscopy to evaluate cellular architecture before and after infection. Mono- and dual-cell cultures were infected with the laboratory strain\n                      <jats:italic>Pseudomonas aeruginosa<\/jats:italic>\n                      PAO1.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Macrophages adhered stably to the basolateral surface without compromising epithelial barrier integrity. Following infection, IL-8 secretion was elevated in epithelial monocultures compared to dual-cell cultures, suggesting early immune modulation in the presence of macrophages. While overall bacterial burden was comparable, confocal imaging revealed clustered bacterial growth in monocultures and a more dispersed spatial distribution in dual-cell cultures.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>This dual-cell ALI model enables investigation of early epithelial\u2013immune interactions, inflammatory modulation, and bacterial colonization dynamics during airway infection. The system provides a versatile and human-relevant platform for studying respiratory host\u2013pathogen interactions.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fcimb.2026.1788554","type":"journal-article","created":{"date-parts":[[2026,4,10]],"date-time":"2026-04-10T04:27:55Z","timestamp":1775795275000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Host-pathogen-immune interactions in an air-liquid interface airway model"],"prefix":"10.3389","volume":"16","author":[{"given":"Alexander F.","family":"Melanson","sequence":"first","affiliation":[{"name":"Department of Clinical Microbiology 9301","place":["Rigshospitalet, Copenhagen, Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Annika","family":"Hettich","sequence":"additional","affiliation":[{"name":"Department of Clinical Microbiology 9301","place":["Rigshospitalet, Copenhagen, Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudia Antonella","family":"Colque","sequence":"additional","affiliation":[{"name":"Department of Clinical Microbiology 9301","place":["Rigshospitalet, Copenhagen, Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jenny J.","family":"Persson","sequence":"additional","affiliation":[{"name":"Department of Experimental Medical Science, Section for Immunology, Lund University","place":["Lund, Sweden"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pablo","family":"Laborda","sequence":"additional","affiliation":[{"name":"Department of Clinical Microbiology 9301","place":["Rigshospitalet, Copenhagen, Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Signe","family":"Lolle","sequence":"additional","affiliation":[{"name":"Department of Clinical Microbiology 9301","place":["Rigshospitalet, Copenhagen, Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S\u00f8ren","family":"Molin","sequence":"additional","affiliation":[{"name":"The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark","place":["Lyngby, Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Helle Krogh","family":"Johansen","sequence":"additional","affiliation":[{"name":"Department of Clinical Microbiology 9301","place":["Rigshospitalet, Copenhagen, Denmark"]},{"name":"Department of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen","place":["Copenhagen, Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,4,10]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"782","DOI":"10.1159\/000534872","article-title":"The intersection between bacterial metabolism and innate immunity","volume":"15","author":"Acosta","year":"2023","journal-title":"J. Innate Immun."},{"key":"B2","doi-asserted-by":"publisher","first-page":"4","DOI":"10.1002\/anbr.202000111","article-title":"Air-liquid interface cultures of the healthy and diseased human respiratory tract: promises, challenges and future directions","volume":"1","author":"Baldassi","year":"2021","journal-title":"Adv. NanoBiomed Res."},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.3390\/life12081216","article-title":"Novel 3D-printed cell culture inserts for air\u2013liquid interface cell culture","volume":"12","author":"Bauer","year":"2022","journal-title":"Life."},{"key":"B4","doi-asserted-by":"publisher","first-page":"131","DOI":"10.1016\/j.micpath.2017.05.028","article-title":"Expression of CXCR1 (IL-8 receptor A) in splenic, peritoneal macrophages and resident bone marrow cells after acute live or heat killed Staphylococcus aureus stimulation in mice","volume":"109","author":"Bishayi","year":"2017","journal-title":"Microb. Pathogen."},{"key":"B5","doi-asserted-by":"publisher","first-page":"109","DOI":"10.1016\/j.cellimm.2010.12.004","article-title":"Characterization of monocyte maturation\/differentiation that facilitates their transmigration across the blood-brain barrier and infection by HIV: implications for NeuroAIDS","volume":"267","author":"Buckner","year":"2011","journal-title":"Cell. Immunol."},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.3390\/toxics13121065","article-title":"Co-culture of primary human bronchial epithelial cells at the air\u2013liquid interface and THP-1 macrophages to investigate the toxicity of polycyclic aromatic hydrocarbons","volume":"13","author":"Burns","year":"2025","journal-title":"Toxics"},{"key":"B7","doi-asserted-by":"publisher","first-page":"411","DOI":"10.1016\/j.cell.2020.08.048","article-title":"Macrophages maintain epithelium integrity by limiting fungal product absorption","volume":"183","author":"Chikina","year":"2020","journal-title":"Cell."},{"key":"B8","doi-asserted-by":"publisher","first-page":"621","DOI":"10.1038\/s41579-022-00682-4","article-title":"Tolerance and resistance of microbial biofilms","volume":"20","author":"Ciofu","year":"2022","journal-title":"Nat. Rev. Microbiol."},{"key":"B9","doi-asserted-by":"publisher","first-page":"1275","DOI":"10.1007\/s00011-023-01743-x","article-title":"Biofilm-forming strains of P. aeruginosa and S. aureus isolated from cystic fibrosis patients differently affect inflammatory phenotype of macrophages","volume":"72","author":"Ciszek-Lenda","year":"2023","journal-title":"Inflammation Res."},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2026.116951","article-title":"Cell-type-independent infection dynamics of clinical Pseudomonas aeruginosa isolates in human airway epithelial models","volume":"45","author":"Colque","year":"2026","journal-title":"Cell Rep."},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1128\/msphere.00916-19","article-title":"Dual and triple epithelial coculture model systems with donor-derived microbiota and THP-1 macrophages to mimic host-microbe interactions in the human sinonasal cavities","volume":"5","author":"De Rudder","year":"2020","journal-title":"mSphere"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1186\/s12950-016-0117-4","article-title":"Mycobacterium leprae alters classical activation of human monocytes in vitro","volume":"13","author":"Fallows","year":"2016","journal-title":"J. Inflammation (United Kingdom)"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1183\/13993003.02399-2016","article-title":"Human bronchial epithelium orchestrates dendritic cell activation in severe asthma","volume":"49","author":"Gras","year":"2017","journal-title":"Eur. Respir. J."},{"key":"B14","doi-asserted-by":"publisher","first-page":"5460","DOI":"10.1182\/bloodadvances.2020002683","article-title":"The transition of M-CSF\u2013derived human macrophages to a growth-promoting phenotype","volume":"4","author":"Hamidzadeh","year":"2020","journal-title":"Blood Adv."},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0279037","article-title":"Expanded characterization of in vitro polarized M0, M1, and M2 human monocyte-derived macrophages: Bioenergetic and secreted mediator profiles","volume":"18","author":"Hickman","year":"2023","journal-title":"PLoS One"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.29132.001","article-title":"\u2018Bacterial colonization stimulates a complex physiological response in the immature human intestinal epithelium\u2019","author":"Hill","year":"2017"},{"key":"B17","doi-asserted-by":"publisher","first-page":"526","DOI":"10.1038\/d41586-025-03218-x","article-title":"The rise of \u201cnightmare bacteria\u201d: antimicrobial resistance in five charts","volume":"646","author":"Kavanagh","year":"2025","journal-title":"Nature"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-024-46938-w","article-title":"Mutations in the efflux pump regulator MexZ shift tissue colonization by Pseudomonas aeruginosa to a state of antibiotic tolerance","volume":"15","author":"Laborda","year":"2024","journal-title":"Nat. Commun."},{"key":"B19","doi-asserted-by":"publisher","first-page":"1076","DOI":"10.1016\/j.tim.2025.05.004","article-title":"Pseudomonas aeruginosa: persistence beyond antibiotic resistance","volume":"33","author":"La Rosa","year":"2025","journal-title":"Trends Microbiol."},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2020.611133","article-title":"Activated platelets convert CD14+CD16- into CD14+CD16+ monocytes with enhanced Fc\u03b3R-mediated phagocytosis and skewed M2 polarization","volume":"11","author":"Lee","year":"2020","journal-title":"Front. Immunol."},{"key":"B21","doi-asserted-by":"publisher","first-page":"1725","DOI":"10.1038\/s41564-024-01718-6","article-title":"Pseudomonas aeruginosa breaches respiratory epithelia through goblet cell invasion in a microtissue model","volume":"9","author":"Leoni Swart","year":"2024","journal-title":"Nat. Microbiol."},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1002\/advs.202511955","article-title":"Cyclic strain and macrophage-mediated transport govern micron-sized PM2.5 translocation across the air\u2013blood barrier","volume":"6","author":"Li","year":"2025","journal-title":"Adv. Sci"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2020.618414","article-title":"The macrophage response to Mycobacterium tuberculosis and opportunities for autophagy inducing nanomedicines for tuberculosis therapy","volume":"10","author":"Maphasa","year":"2020","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"B24","doi-asserted-by":"publisher","first-page":"228","DOI":"10.1016\/j.ajpath.2020.10.017","article-title":"Animal and cell culture models for cystic fibrosis: Which model is right for your application","volume":"191","author":"McCarron","year":"2021","journal-title":"Am. J. Pathol."},{"key":"B25","doi-asserted-by":"publisher","first-page":"443","DOI":"10.1007\/s00441-011-1301-8","article-title":"Transmigration of macrophages across the choroid plexus epithelium in response to the feline immunodeficiency virus","volume":"347","author":"Meeker","year":"2012","journal-title":"Cell Tissue Res."},{"key":"B26","doi-asserted-by":"publisher","first-page":"e955","DOI":"10.1016\/S2468-2667(25)00201-4","article-title":"The burden of bacterial antimicrobial resistance in the WHO Eastern Mediterranean Region 1990\u20132021: a cross-country systematic analysis with forecasts to 2050","volume":"10","author":"Mestrovic","year":"2025","journal-title":"Lancet Public Health"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.3390\/ijms23169125","article-title":"RNA-Seq analysis of UPM-exposed epithelium co-cultivated with macrophages and dendritic cells in obstructive lung diseases","volume":"23","author":"Misiukiewicz-St\u0119pien","year":"2022","journal-title":"Int. J. Mol. Sci."},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1128\/microbiolspec.mchd-0012-2015","article-title":"Strategies used by bacteria to grow in macrophages","volume":"4","author":"Mitchell","year":"2016","journal-title":"Microbiol. Spectr."},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.3390\/cells13242068","article-title":"A brief history of cell culture: From Harrison to organs-on-a-chip","volume":"13","author":"Moro","year":"2024","journal-title":"Cells"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2021.625597","article-title":"Immune responses to Pseudomonas aeruginosa biofilm infections","volume":"12","author":"Moser","year":"2021","journal-title":"Front. Immunol."},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2024.1405376","article-title":"Pseudomonas aeruginosa in chronic lung disease: untangling the dysregulated host immune response","volume":"15","author":"Nickerson","year":"2024","journal-title":"Front. Immunol."},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.0030193","article-title":"Hemophagocytic macrophages harbor Salmonella enterica during persistent infection","volume":"3","author":"Nix","year":"2007","journal-title":"PLoS Pathog."},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-021-92626-w","article-title":"Interactions of nasal epithelium with macrophages and dendritic cells variously alter urban PM-induced inflammation in healthy, asthma and COPD","volume":"11","author":"Paplinska-Goryca","year":"2021","journal-title":"Sci. Rep."},{"key":"B34","doi-asserted-by":"publisher","first-page":"192","DOI":"10.1038\/s42003-024-05867-6","article-title":"Akkermansia muciniphila-induced trained immune phenotype increases bacterial intracellular survival and attenuates inflammation","volume":"7","author":"Pe\u00f1a-Cearra","year":"2024","journal-title":"Commun. Biol."},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1186\/s12866-022-02623-w","article-title":"A microfluidic-based analysis of 3D macrophage migration after stimulation by Mycobacterium, Salmonella and Escherichia","volume":"22","author":"P\u00e9rez-Rodr\u00edguez","year":"2022","journal-title":"BMC Microbiol."},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1186\/s12931-023-02514-2","article-title":"A comparative study of in vitro air\u2013liquid interface culture models of the human airway epithelium evaluating cellular heterogeneity and gene expression at single cell resolution","volume":"24","author":"Prescott","year":"2023","journal-title":"Respir. Res."},{"key":"B37","doi-asserted-by":"publisher","first-page":"331","DOI":"10.1038\/s41579-020-00477-5","article-title":"Pseudomonas aeruginosa adaptation and evolution in patients with cystic fibrosis","volume":"19","author":"Rossi","year":"2021","journal-title":"Nat. Rev. Microbiol."},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1080\/19490976.2024.2409924","article-title":"Helicobacter pylori PldA modulates TNFR1-mediated p38 signaling pathways to regulate macrophage responses for its survival","volume":"16","author":"Sit","year":"2024","journal-title":"Gut Microbes"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.1183\/16000617.0172-2023","article-title":"A narrative review of chemokine receptors CXCR1 and CXCR2 and their role in acute respiratory distress syndrome","volume":"33","author":"Toya","year":"2024","journal-title":"Eur. Respir. Rev"},{"key":"B40","unstructured":"Generation of a human airway epithelium derived basal cell line with multipotent differentiation capacity\n          \n          \n            \n              Walters\n              M. S.\n            \n            \n              Kuzel\n              T. M.\n            \n            \n              Shafikhani\n              S. H.\n            \n          \n          \n          2013"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.3390\/cells12010199","article-title":"Pseudomonas aeruginosa: infections, animal modeling, and therapeutics","volume":"12","author":"Wood","year":"2023","journal-title":"Cells"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.18063\/ijb.755","article-title":"Pore size-mediated macrophage M1 to M2 transition affects osseointegration of 3D-printed PEEK scaffolds","volume":"9","author":"Yang","year":"2023","journal-title":"Int. J. Bioprinting"}],"container-title":["Frontiers in Cellular and Infection Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2026.1788554\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,10]],"date-time":"2026-04-10T04:27:57Z","timestamp":1775795277000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2026.1788554\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,10]]},"references-count":42,"alternative-id":["10.3389\/fcimb.2026.1788554"],"URL":"https:\/\/doi.org\/10.3389\/fcimb.2026.1788554","relation":{},"ISSN":["2235-2988"],"issn-type":[{"value":"2235-2988","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,10]]},"article-number":"1788554"}}