Host-pathogen-immune interactions in an air-liquid interface airway model
(2026) In Frontiers in cellular and infection microbiology 16.- 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... (More)
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.
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- author
- Melanson, Alexander F. ; Hettich, Annika ; Colque, Claudia Antonella ; Persson, Jenny J. LU ; Laborda, Pablo ; Lolle, Signe ; Molin, Søren and Johansen, Helle Krogh
- organization
- publishing date
- 2026
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- air-liquid interface (ALI), airway infection model, host-pathogen interaction, infection model, macrophages, Pseudomonas aeruginosa
- in
- Frontiers in cellular and infection microbiology
- volume
- 16
- article number
- 1788554
- publisher
- Frontiers Media S. A.
- external identifiers
-
- scopus:105037096422
- pmid:42039749
- ISSN
- 2235-2988
- DOI
- 10.3389/fcimb.2026.1788554
- language
- English
- LU publication?
- yes
- id
- 7514d796-4926-4e85-ad29-3d20266332cb
- date added to LUP
- 2026-06-23 14:07:39
- date last changed
- 2026-08-05 23:16:35
@article{7514d796-4926-4e85-ad29-3d20266332cb,
abstract = {{<p>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.</p>}},
author = {{Melanson, Alexander F. and Hettich, Annika and Colque, Claudia Antonella and Persson, Jenny J. and Laborda, Pablo and Lolle, Signe and Molin, Søren and Johansen, Helle Krogh}},
issn = {{2235-2988}},
keywords = {{air-liquid interface (ALI); airway infection model; host-pathogen interaction; infection model; macrophages; Pseudomonas aeruginosa}},
language = {{eng}},
publisher = {{Frontiers Media S. A.}},
series = {{Frontiers in cellular and infection microbiology}},
title = {{Host-pathogen-immune interactions in an air-liquid interface airway model}},
url = {{http://dx.doi.org/10.3389/fcimb.2026.1788554}},
doi = {{10.3389/fcimb.2026.1788554}},
volume = {{16}},
year = {{2026}},
}