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Toxicological Sciences 60, 356-362 (2001)
Copyright © 2001 by the Society of Toxicology

RESPIRATORY TOXICOLOGY

Effects of Ozone Exposure on Nuclear Factor-{kappa}B Activation and Tumor Necrosis Factor-{alpha} Expression in Human Nasal Epithelial Cells

Brian G. Nichols, James S. Woods, Daniel L. Luchtel, Jeannette Corral and Jane Q. Koenig,1

Department of Environmental Health, University of Washington, Seattle, Washington

In this study we investigated a possible mechanism of the human airway inflammatory response to inhaled ozone (O3). Cultures of human nasal epithelial (HNE) cells, initiated from excised nasal turbinates and grown on collagen-coated Transwell® tissue culture inserts, were exposed to 120, 240, or 500 ppb O3 for 3 h. An electron spin resonance (ESR) signal that changed with time suggested free radical production in HNE cells exposed to O3. Nuclear protein extracts were analyzed for the activated transcription factor NF-{kappa}B by electrophoretic mobility-shift assay (EMSA), and showed a small dose-response activation of NF-{kappa}B that coincided with O3-induced free radical production. Basal media were analyzed for the presence of tumor necrosis factor-{alpha} (TNF-{alpha}) using the enzyme-linked immunosorbent assay (ELISA). In cultures exposed to 120 ppb O3, the mean TNF-{alpha} concentration was not significantly different from those exposed to air. However, exposure to 240 and 500 ppb O3 significantly increased mean TNF-{alpha} expression, relative to controls, 16 h after exposure. These results support the hypothesis that the human airway epithelium plays a role in directing the inflammatory response to inhaled O3 via free radical-mediated NF-{kappa}B activation.

Key Words: airway; cell culture; cytokine; electron spin resonance; epithelial cell, free radical; inflammation; nasal cell; ozone; transcription factor; tumor necrosis factor..


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