Interactions between human bronchoepithelial cells and lung fibroblasts after ozone exposure in vitro

Long-term exposure to ozone has been shown to cause lung fibrosis and increased collagen synthesis by fibroblasts in experimental animals. As the bronchial epithelium appears to play a major regulatory role in inflammatory processes, we investigated whether ozone induces bronchoepithelial cells in vitro to increase gene expression of procollagens and other fibrogenic mediators in human lung fibroblasts. Membrane cultures of human airway epithelial cells (BEAS-2B) in the presence or absence of lung fibroblast (HFL-1) cultures were exposed to air or 500 ppb ozone for 1 h, followed by (co-)incubation periods of 11 and 23 h. After ozone exposure of the co-cultures, there were substantial increases of steady-state mRNA levels of both alpha1 procollagens type I and III as well as TGF beta1 in the fibroblasts above the corresponding air control levels. In the absence of ozone, the presence of epithelial cells always caused significant decreases in the basal steady-state mRNA levels of both procollagens as compared to their absence. There were no significant effects of ozone on the secretion or gene expression of TGF beta2, PDGF or IL-8 in any cell type. In contrast, co-culture condition induced altered patterns of IL-8 gene expression or of PDGF production in fibroblasts and bronchoepithelial cells, respectively, both in the absence or presence of ozone. In summary, our data demonstrate that the effect of ozone on fibroblasts was mediated by epithelial cells and that mutual regulatory interactions between the different cell types occur. Thus, our co-cultivation system in vitro appears to be able to mimic the in vivo situation providing insight into the nature of cellular interactions and modulation by ozone, which may occur in the whole organism after long-term exposure.