synthesis of PAl-I, and an increase in PA inhibitory activity. A more complete understanding of the regulation of epithelial cell u-PAIPAI-1 expression may provide insight into repair of lung injury and potential ways to favorably manipulate this process. REFERENCES 1 Saksela 0, Rifkin DB. Cell associated plasminogen activation: regulation and physiologic functions. Ann Rev Cell Bioi 1988; 4:93-126 2 Idell S, Peters J, James KK, Fair DS, Coalson JJ. Local abnormalities in coagulation and fibrinolytic pathways predispose to alveolar fibrin deposition in adult respiratory distress syndrome. J Clin Invest 1989; 84:181-93 3 Bertozzi
~
Astedt B, Zenzius L, Lynch K, LeMaire F, Zapol ~
et a1. Depressed bronchoalveolar urokinase activity in patients with adult respiratory distress syndrome. N Engl J Med 1990; 322:890-97 4 Gerwin BI, Keski-Oja J, Seddon M, Lechner JF, Harris CC. TGF-~1
modulation of urokinase and PAI-l expression in human
bronchial epithelial cells. Am J Physioll990; 259:262-69 5 Marshall BC, Sageser DS, Rao
N~
Emi M, Hoidal JR. Alveolar
epithelial cell plasminogen activator: characterization and regulation. J Bioi Chern 1990; 265:8198-8204
6 Gross TJ, Simon RH, Kelly CJ, Sitrin RG. Rat alveolar epithelial cells concomitantly express plasminogen activator inhibitor-l and urokinase. Am J Physioll991; 260:286-95
7 Keski-Oja J, Blasi F, Leof EB, Moses HL. Regulation of the synthesis and activity of urokinase plasminogen activator in A549 human lung carcinoma cells by transforming growth Cell Bioi 1988; 106:451-59 8 Marshall BC, Xu
Q-~
Rao
N~
factor-~.
J
Brown BR, Hoidal JR. Pulmonary
epithelial cell urokinase type plasminogen activator: induction by
IL-l~
and TNF-a (submitted)
9 Shimada H, Mori T, Takada A, Takada Y, Noda Y, Takai I, et a1. Use of chromogenic substrate S-2251 for determination of plasminogen activator in rat ovaries. Thromb Haemostasis 1981; 46:507-16
228
Cultured Human Bronchial Epithelial Cells Release Cytokines, Fibronectin, and Lipids in Response to Ozone Exposure* Karen McKinnon, Ph.D.; L Noah, M.D.; M. Madden, Ph.D.; H. Koren, Ph.D.; and R. Devlin, Ph.D.
exposed the bronchial epithelial cell line, BEAS-2B W e(Reddel R, et al. Can Res 1988; 48:19(4), cultured on
collagen-impregnated filters to air or 0.1, 0.25, 0.5, or 1.0 ppm ozone (03) for 1 h. Fluid obtained from either apical or basolateral compartments was analyzed for cytokines and inflammatory mediators released in response to 0 3 exposure. The 0 3 had dose-dependent cytotoxic effects as determined by the release of LDH or 51Cr and by the uptake of trypan blue or propidium iodide. Fibronectin (ELISA) and IL-8 (RIA) appeared in both apical and basolateral compartments within 1 h following exposure to concentrations as low as 0.1 ppm 03' The PAF (RIA) appeared in apical and basolateral compartments within minutes following exposure to 03' Exposure of BEAS2B cells to 0.1 to 1.0 ppm 0 3 caused the vectorial release of IL-6 into the apical compartment within 4 h as measured in bioassay and ELISA. IL 6 could be measured in the basolateral compartment within 24 h. Low concentrations of 0 3 (0.1 ppm) caused a rapid release of PGE 2 and TXB2 (RIA) in both the apical and basolateral compartments of BEAS-2B cells. Higher concentrations of 0 3 stimulated dose-dependent release of additional arachidonic acid metabolites. The HPLC analyses by two different separation methods showed the presence of peaks that comigrated with authentic standards for TXB 2 , PGE 2 , PGF2Q, LTC.., LTD.., I5-HETE, and HHT. Thus, 0 3 stimulated the BEAS-2B cells to release many of the same inflammatory mediators that are detected in the bronchoalveolar lavage fluid of humans exposed to 0 3 (Koren H, et al. Arn Rev Respir Dis 1989; 139:407) which indicates that this model may be useful in the study of human airway inflammation. *From the Health Effects Research Laboratory, US Environmental Protection Agency, Research Triangle Park, N.C. This abstract ofa proposed presentation does not necessarily reflect EPA policy.
34th Annual Thomas L Petty Aspen Lung Conference
~
Astedt B, Zenzius L, Lynch K, LeMaire F, Zapol ~
et a1. Depressed bronchoalveolar urokinase activity in patients with adult respiratory distress syndrome. N Engl J Med 1990; 322:890-97 4 Gerwin BI, Keski-Oja J, Seddon M, Lechner JF, Harris CC. TGF-~1
modulation of urokinase and PAI-l expression in human
bronchial epithelial cells. Am J Physioll990; 259:262-69 5 Marshall BC, Sageser DS, Rao
N~
Emi M, Hoidal JR. Alveolar
epithelial cell plasminogen activator: characterization and regulation. J Bioi Chern 1990; 265:8198-8204
6 Gross TJ, Simon RH, Kelly CJ, Sitrin RG. Rat alveolar epithelial cells concomitantly express plasminogen activator inhibitor-l and urokinase. Am J Physioll991; 260:286-95
7 Keski-Oja J, Blasi F, Leof EB, Moses HL. Regulation of the synthesis and activity of urokinase plasminogen activator in A549 human lung carcinoma cells by transforming growth Cell Bioi 1988; 106:451-59 8 Marshall BC, Xu
Q-~
Rao
N~
factor-~.
J
Brown BR, Hoidal JR. Pulmonary
epithelial cell urokinase type plasminogen activator: induction by
IL-l~
and TNF-a (submitted)
9 Shimada H, Mori T, Takada A, Takada Y, Noda Y, Takai I, et a1. Use of chromogenic substrate S-2251 for determination of plasminogen activator in rat ovaries. Thromb Haemostasis 1981; 46:507-16
228
Cultured Human Bronchial Epithelial Cells Release Cytokines, Fibronectin, and Lipids in Response to Ozone Exposure* Karen McKinnon, Ph.D.; L Noah, M.D.; M. Madden, Ph.D.; H. Koren, Ph.D.; and R. Devlin, Ph.D.
exposed the bronchial epithelial cell line, BEAS-2B W e(Reddel R, et al. Can Res 1988; 48:19(4), cultured on
collagen-impregnated filters to air or 0.1, 0.25, 0.5, or 1.0 ppm ozone (03) for 1 h. Fluid obtained from either apical or basolateral compartments was analyzed for cytokines and inflammatory mediators released in response to 0 3 exposure. The 0 3 had dose-dependent cytotoxic effects as determined by the release of LDH or 51Cr and by the uptake of trypan blue or propidium iodide. Fibronectin (ELISA) and IL-8 (RIA) appeared in both apical and basolateral compartments within 1 h following exposure to concentrations as low as 0.1 ppm 03' The PAF (RIA) appeared in apical and basolateral compartments within minutes following exposure to 03' Exposure of BEAS2B cells to 0.1 to 1.0 ppm 0 3 caused the vectorial release of IL-6 into the apical compartment within 4 h as measured in bioassay and ELISA. IL 6 could be measured in the basolateral compartment within 24 h. Low concentrations of 0 3 (0.1 ppm) caused a rapid release of PGE 2 and TXB2 (RIA) in both the apical and basolateral compartments of BEAS-2B cells. Higher concentrations of 0 3 stimulated dose-dependent release of additional arachidonic acid metabolites. The HPLC analyses by two different separation methods showed the presence of peaks that comigrated with authentic standards for TXB 2 , PGE 2 , PGF2Q, LTC.., LTD.., I5-HETE, and HHT. Thus, 0 3 stimulated the BEAS-2B cells to release many of the same inflammatory mediators that are detected in the bronchoalveolar lavage fluid of humans exposed to 0 3 (Koren H, et al. Arn Rev Respir Dis 1989; 139:407) which indicates that this model may be useful in the study of human airway inflammation. *From the Health Effects Research Laboratory, US Environmental Protection Agency, Research Triangle Park, N.C. This abstract ofa proposed presentation does not necessarily reflect EPA policy.
34th Annual Thomas L Petty Aspen Lung Conference
