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Am J Physiol Lung Cell Mol Physiol 278: L25-L32, 2000;
1040-0605/00 $5.00
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Vol. 278, Issue 1, L25-L32, January 2000

Mechanisms of GM-CSF increase by diesel exhaust particles in human airway epithelial cells

Sonja Boland1, Véronique Bonvallot1, Thierry Fournier2, Armelle Baeza-Squiban1, Michel Aubier2, and Francelyne Marano1

1 Laboratoire de Cytophysiologie et Toxicologie Cellulaire, Université Paris VII, 75251 Paris; and 2 Institut National de la Santé et de la Recherche Médicale Unité 408, Faculté X. Bichat, 75018 Paris, France

We have previously shown that exposure to diesel exhaust particles (DEPs) stimulates human airway epithelial cells to secrete the inflammatory cytokines interleukin-8, interleukin-1beta , and granulocyte-macrophage colony-stimulating factor (GM-CSF) involved in allergic diseases. In the present paper, we studied the mechanisms underlying the increase in GM-CSF release elicited by DEPs using the human bronchial epithelial cell line 16HBE14o-. RT-PCR analysis has shown an increase in GM-CSF mRNA levels after DEP treatments. Comparison of the effects of DEPs, extracted DEPs, or extracts of DEPs has shown that the increase in GM-CSF release is mainly due to the adsorbed organic compounds and not to the metals present on the DEP surface because the metal chelator desferrioxamine had no inhibitory effect. Furthermore, radical scavengers inhibited the DEP-induced GM-CSF release, showing involvement of reactive oxygen species in this response. Moreover genistein, a tyrosine kinase inhibitor, abrogated the effects of DEPs on GM-CSF release, whereas protein kinase (PK) C, PKA, cyclooxygenase, or lipoxygenase inhibitors had no effect. PD-98059, an inhibitor of mitogen-activated protein kinase, diminished the effects of DEPs, whereas SB-203580, an inhibitor of p38 mitogen-activated protein kinase, had a lower effect, and DEPs did actually increase the active, phosphorylated form of the extracellular signal-regulated kinase as shown by Western blotting. In addition, cytochalasin D, which inhibits the phagocytosis of DEPs, reduced the increase in GM-CSF release after DEP treatment. Together, these data suggest that the increase in GM-CSF release is mainly due to the adsorbed organic compounds and that the effect of native DEPs requires endocytosis of the particles. Reactive oxygen species and tyrosine kinase(s) may be involved in the DEP-triggered signaling of the GM-CSF response.

granulocyte-macrophage colony-stimulating factor; 16HBE14o- cells; signal transduction; reactive oxygen species; cytokines; inflammation


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