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Am. J. Respir. Cell Mol. Biol., Volume 26, Number 4, April, 2002 499-505

Lung Macrophage-Epithelial Cell Interactions Amplify Particle-Mediated Cytokine Release

Florence Tao and Lester Kobzik

Department of Environmental Health, Harvard School of Public Health, Boston, Massachusetts; and Department of Pathology, Brigham and Women's Hospital, Boston, Massachusetts

Interactions between alveolar macrophages (AMs) and epithelial cells may promote inflammatory responses to air pollution particles. Normal rat AMs, the alveolar type II epithelial cell line RLE-6TN (RLE), or cocultures of both cell types were incubated with various particles (0-50 µg/ml) for 24 h, followed by assay of released TNF-alpha and MIP-2. The particles used included titanium dioxide (TiO2), alpha -quartz (SiO2), residual oil fly ash (ROFA), or urban air particles (UAP). For all particles, a dose-dependent increase in TNF-alpha and MIP-2 release was observed in AM+RLE co-cultures but not in RLE or AM monoculture. AM+RLE co-culture also synergistically enhanced basal levels of tumor necrosis factor (TNF)-alpha and macrophage inflammatory protein (MIP)-2. In contrast, when AMs were co-cultured with fibroblasts, basal and particle-induced TNF-alpha and MIP-2 were similar to levels found in AM monoculture. Particle uptake by AMs was similar in mono- or AM+RLE co-culture. Increased basal and particle-induced cytokine release were not observed when the AMs were physically separated from the RLE. This contact-dependent cytokine potentiation could not be blocked with anti-CD18/anti-CD54, arginine-glycine-aspartate (RGD) peptide, or heparin. We conclude that in vitro inflammatory responses to particles are amplified by contact-dependent interactions between AMs and epithelial cells. AM-epithelial co-culture may provide a useful model of in vivo particle effects.

Abbreviations: alveolar macrophage, AM; bronchoalveolar lavage, BAL; Dulbecco's modified Eagle's medium, DMEM; fetal bovine serum, FBS; interleukin, IL; lipopolysaccharide, LPS; macrophage inflammatory protein, MIP; right angle scatter, RAS; recombinant endotoxin neutralizing protein, rENP; arginine-glycine-aspartate, RGD; RLE-6TN epithelial cells, RLE; residual oil fly ash, ROFA; alpha -quartz, SiO2; titanium dioxide, TiO2; tumor necrosis factor, TNF.




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