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Am. J. Respir. Cell Mol. Biol., Volume 25, Number 3, September, 2001 265-271

Particulate Matter Induces Cytokine Expression in Human Bronchial Epithelial Cells

Takeshi Fujii, Shizu Hayashi, James C. Hogg, Renaud Vincent,* and Stephan F. Van Eeden

Pulmonary Research Laboratory, University of British Columbia, St. Paul's Hospital, Vancouver, British Columbia; and * Directorate of Environmental Health, Health Canada, Ottawa, Ontario, Canada

The present study was designed to determine cytokines produced by primary human bronchial epithelial cells (HBECs) exposed to ambient air pollution particles (EHC-93). Cytokine messenger RNA (mRNA) was measured using a ribonuclease protection assay and cytokine protein production by enzyme-linked immunosorbent assay. Primary HBECs were freshly isolated from operated lung, cultured to confluence, and exposed to 10 to 500 µg/ml of a suspension of ambient particulate matter with a diameter of less than 10 µm (PM10) for 2, 8, and 24 h. The mRNA levels of leukemia inhibitory factor (LIF), granulocyte macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-1alpha , and IL-8 were increased after exposure to PM10, and this increase was dose-dependent between 100 (P < 0.05) and 500 (P < 0.05) µg/ml of PM10 exposure. The concentrations of LIF, GM-CSF, IL-1beta , and IL-8 protein measured in the supernatant collected at 24 h increased in a dose- dependent manner and were significantly higher than those in the control nonexposed cells. The soluble fraction of the PM10 (100 µg/ml) did not increase these cytokine mRNA levels compared with control values and were significantly lower compared with HBECs exposed to 100 µg/ml of PM10 (LIF, IL-8, and IL-1beta ; P < 0.05), except for GM-CSF mRNA (P = not significant). We conclude that primary HBECs exposed to ambient PM10 produce proinflammatory mediators that contribute to the local and systemic inflammatory response, and we speculate that these mediators may have a role in the pathogenesis of cardiopulmonary disease associated with particulate air pollution.

Abbreviations: antibody, Ab; bronchial epithelial cell growth medium, BEGM; colloidal carbon, CC; diesel exhaust particles, DEP; enzyme-linked immunosorbent assay, ELISA; glyceraldehyde-3-phosphate dehydrogenase, GAPDH; granulocyte-CSF, G-CSF; granulocyte macrophage colony-stimulating factor, GM-CSF; human bronchial epithelial cell, HBEC; interleukin, IL; leukemia inhibitory factor, LIF; lipopolysaccharide, LPS; monocyte chemotactic protein, MCP; macrophage-CSF, M-CSF; messenger RNA, mRNA; particulate matter with a diameter of less than 10 µm, PM10; regulated on activation, normal T cells expressed and secreted, RANTES; ribonuclease protection assay, RPA; standard error of the mean, SEM; tumor necrosis factor, TNF.




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