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Host-pathogen-immune interactions in an air-liquid interface airway model.

Title: Host-pathogen-immune interactions in an air-liquid interface airway model.
Authors: Melanson AF; Department of Clinical Microbiology 9301, Rigshospitalet, Copenhagen, Denmark.; Hettich A; Department of Clinical Microbiology 9301, Rigshospitalet, Copenhagen, Denmark.; Colque CA; Department of Clinical Microbiology 9301, Rigshospitalet, Copenhagen, Denmark.; Persson JJ; Department of Experimental Medical Science, Section for Immunology, Lund University, Lund, Sweden.; Laborda P; Department of Clinical Microbiology 9301, Rigshospitalet, Copenhagen, Denmark.; Lolle S; Department of Clinical Microbiology 9301, Rigshospitalet, Copenhagen, Denmark.; Molin S; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Lyngby, Denmark.; Johansen HK; Department of Clinical Microbiology 9301, Rigshospitalet, Copenhagen, Denmark.; Department of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
Source: Frontiers in cellular and infection microbiology [Front Cell Infect Microbiol] 2026 Apr 10; Vol. 16, pp. 1788554. Date of Electronic Publication: 2026 Apr 10 (Print Publication: 2026).
Publication Type: Journal Article
Language: English
Journal Info: Publisher: Frontiers Media SA Country of Publication: Switzerland NLM ID: 101585359 Publication Model: eCollection Cited Medium: Internet ISSN: 2235-2988 (Electronic) Linking ISSN: 22352988 NLM ISO Abbreviation: Front Cell Infect Microbiol Subsets: MEDLINE
Imprint Name(s): Original Publication: Lausanne : Frontiers Media SA
MeSH Terms: Host-Pathogen Interactions*/immunology ; Pseudomonas aeruginosa*/immunology ; Pseudomonas aeruginosa*/growth & development ; Macrophages*/immunology ; Macrophages*/microbiology ; Epithelial Cells*/microbiology ; Epithelial Cells*/immunology; Interleukin-8/metabolism ; Pseudomonas Infections/immunology ; Pseudomonas Infections/microbiology ; Respiratory Mucosa/immunology ; Respiratory Mucosa/microbiology ; Cell Culture Techniques/methods ; Humans ; Cells, Cultured ; Microscopy, Confocal
Abstract: Background: Air-liquid 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-immune crosstalk. To address this, we developed a dual-cell ALI model incorporating human peripheral blood monocyte-derived macrophages beneath differentiated airway epithelial cells.; Methodology: 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 Pseudomonas aeruginosa PAO1.; Results: 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.; Conclusions: This dual-cell ALI model enables investigation of early epithelial-immune interactions, inflammatory modulation, and bacterial colonization dynamics during airway infection. The system provides a versatile and human-relevant platform for studying respiratory host-pathogen interactions.; (Copyright © 2026 Melanson, Hettich, Colque, Persson, Laborda, Lolle, Molin and Johansen.)
Competing Interests: The author(s) declared that this work was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Contributed Indexing: Keywords: Pseudomonas aeruginosa; air-liquid interface (ALI); airway infection model; host-pathogen interaction; infection model; macrophages
Substance Nomenclature: 0 (Interleukin-8); 0 (CXCL8 protein, human)
Entry Date(s): Date Created: 20260427 Date Completed: 20260427 Latest Revision: 20260427
Update Code: 20260427
PubMed Central ID: PMC13106080
DOI: 10.3389/fcimb.2026.1788554
PMID: 42039749
Database: MEDLINE

Journal Article