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Cytokine-induced airway epithelial ICAM-1 upregulation: quantification by high-resolution scanning and transmission
J Zhu1, A V Rogers, A Burke-Gaffney
1Lung Pathology and Applied Pharmacology, National Heart and Lung Institute, Imperial College School of Medicine, London, UK.
Insights
Intercellular adhesion molecule-1 (ICAM-1) expression varies significantly between airway epithelial cells. Basal and indeterminate cells show higher ICAM-1, suggesting a key role in leukocyte adhesion.
Area of Science:
- Cell Biology
- Immunology
- Respiratory Medicine
Background:
- Intercellular adhesion molecule-1 (ICAM-1) is crucial for leukocyte adhesion in inflammatory responses.
- Airway epithelial cells exhibit diverse phenotypes and functions, potentially influencing ICAM-1 expression.
Purpose of the Study:
- To investigate the variation in ICAM-1 expression among different airway epithelial cell phenotypes.
- To determine the role of specific airway epithelial cell types in ICAM-1-mediated leukocyte adhesion.
Main Methods:
- Cultured human airway epithelial cell lines (16HBE14o- and BEAS-2B) and explants.
- Stimulation with tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ).
- Quantitative analysis using enzyme-linked immunosorbent assay (ELISA), high-resolution scanning electron microscopy (HR-SEM) with immunogold labeling, and immunohistochemistry.
Main Results:
- ELISA showed a 1.6-fold ICAM-1 increase in BEAS-2B cells post-cytokine stimulation, but not in 16HBE cells.
- HR-SEM revealed significant ICAM-1 upregulation in stimulated 16HBE cells (7.9 to 19.1 gold particles/10 μm²).
- BEAS-2B cells exhibited higher constitutive (24.2) and stimulated (68.5) ICAM-1 expression compared to 16HBE cells.
- Immunohistochemistry indicated ICAM-1 expression predominantly on basal and indeterminate epithelial cells in explants.
Conclusions:
- Airway epithelial cells display marked cell-to-cell variation in ICAM-1 expression.
- Basal and indeterminate airway epithelial cells are key players in leukocyte retention due to higher ICAM-1 expression.
- These findings highlight the differential roles of airway epithelial cell subtypes in immune responses.
Abstract:
The aim of this study was to investigate the variation of intercellular adhesion molecule (ICAM)-1 which occurs between individual airway epithelial cells of distinct phenotype. 16HBE14o- (16HBE) and BEAS-2B cell lines and human airway explants were cultured with medium alone or a mixture of tumour necrosis factor (TNF)-alpha (10 ng x mL(-1)) and interferon (IFN)-gamma (40 ng x mL(-1)) before being immunogold-labelled and examined quantitatively using sensitive high-resolution electron microscopic techniques. By enzyme-linked immunosorbent assay there was a 1.6-fold increase of ICAM-1 in the BEAS-2B cells following the cytokine mix which was not apparent in the 16HBE cells. However, high-resolution scanning electron microscopy demonstrated that an upregulation had occurred; median and ranges for gold particle number per 10 microm2 cell surface were: 7.9 (0-40) for nonstimulated and 19.1 (0-60 for stimulated) (p<0.01, Mann-Whitney U-test). The value for the nonstimulated BEAS-2B cells was 24.2 (0-60), 3-times higher that the constitutive expression in the 16HBE cells (p<0.01), whereas following stimulation, it was 68.5 (20-130) (p<0.01). Values for explant epithelial outgrowths were similar to the 16HBE cells. Immunohistochemistry of the explanted mucosa showed both constitutive and upregulated expression of epithelial ICAM-1 associated with basal and indeterminate cells rather than with ciliated or goblet cells. These results using high-resolution techniques indicate that there is marked cell-to-cell variation in cellular adhesion molecule expression and that it is the basal cells and less well differentiated (indeterminate) epithelial cells which are likely to play key roles in leukocyte retention via intercellular adhesion molecule-1.
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