Nat! Acad Sci 1985; 82:4633-37 8 Holtzman MJ. Species speci6city of lipoxygenase and cyclooxygenase activities expressed in pulmonary airway epithelial cells. Adv Prostaglandin Thromhoxane Leukotriene Res 1987; 17:17779 9 Holtzman MJ, Hansborough JR, Rosen GO, Turk J. Uptake, release and novel species-dependent oxygenation of arachidonic acid in human and animal airway epithelial cells. Biochim Biophys Acta 1988; 963:401-13 10 Davies RJ, Toumbis MG, Sapsford RJ, et aI. Functional and metabolic assessment of human bronchial epithelial cells in vitro. J Allergy Clin Immunoll990; 85:155 11 Rosen EM, Goldberg 10. Protein factors which regulate cell motility in vitro. Cell Develop Bioi 1989; 25:1079-87 12 Ruoslahti E, Engvall E, Hayman EH. Fibronectin: current concepts of its structure and functions. Coli Relat Res 1981; 1:95-128 13 Shoji S, Ertl RF, Linder J, Romberger OJ, Rennard SI. Bronchial epithelial ceUs produce chemotactic activity for bronchial epithelial ceUs: possible role for 6bronectin in airway repair. Am Rev Respir Dis 1990; 141:218-25 14 Aas K. Heterogeneity of bronchial asthma. Allergy 1981; 36: 3-10
Expression of the Potent Inflammatory Cytokines, GM-CSF; IL6, and 1L8, in Bronchial Epithelial Cells of Asthmatic Patients· Sabrina Mattoli, M.D., Ph.D.;t* Maurizio Marini, M.D.;* and Angelo Fasoli, M.D. t
C
ultured human bronchial epithelial cells produce cytokines with potent proinflammatory properties upon exposure to immunologic and nonimmunologic stimuli or during recovery from injury!-7 particularly GM-CSF and IL6. Because of this ability to release factors which affect the migration of granulocytes and lymphocytes and promote their local activation,2,3,6-9 airway epithelial cells can be viewed as cells that signal the inflammatory and immune system that penetration or disruption of the airway mucosa has occurred, and epithelial cell-derived cytokines may play an important role in the genesis and persistence of bronchial inflammation in some pathologic conditions, like asthma. Indeed, we have previously demonstrated that the airway secretions from patients with symptomatic asthma contain increased amounts ofGM-CSF and IL6, and these cytokines have been localized in the cytoplasm ofnonciliated bronchial epithelial cells by immunospecific labeling. 10 In addition, we have provided direct evidence of an upregulation of the expression of GM-CSF gene and protein in the bronchial epithelial cells of these patients. 6,7 The GM-CSF released from asthmatic bronchial epithelial cells could prolong the survival of eosinophils, make them hypodense, and promote *From the tSection of Lung Cellular and Molecular Biology, Department of Internal Medicine, University of Milan; and *Diagnostic Center for Respiratory Diseases, Milan, Italy. This study was supported by the Minister of University and Research, Italy. Reprint requests: Dr. Mattoli, Via C. Battisti 5, Tenli, Italy 05100
the release of mediators from those cells,6,7 thereby reproducing in vitro phenomena that are known to occur in asthma and that contribute to the pathogenesis of airway hyperresponsiveness. 11 In the present study, we have further evaluated the expression of cytokines with inflammatory properties in bronchial epithelial cells of asthmatic patients and tested the ability of corticosteroids and nedocromil sodium to downregulate their production. Six asthmatic patients (4 women, mean age ± SD: 31.2±9.8 years) and 6 healthy volunteers (3 women, mean age ± SD: 43.0 ± 15.5 years) completed the study. All subjects were nonsmokers and two in each group showed positive responses to skin prick testing with 12 common allergen extracts 12 (Table 1). The asthmatics were receiving treatment with inhaled bronchodilators alone, on demand, but their symptoms were not controlled, as they were elicited by nonspecific irritating stimuli, or required high doses of bronchodilators daily, or disturbed sleep. No patient had received corticosteroids in the last month. They had pulmonary function measurements and bronchoprovocation tests with methacholine 13 to define their nonspecific bronchial hyperresponsiveness. Their mean baseline FEV. (± SD) was 83.3 ± 14.5% of predicted value) and their geometric mean PCmM was 0.187 mglml (range from 0.04 to 0.844 mwml) (Table 1), the latter indicating that airway reactivity was moderately to severely increased in those patients. Control subjects had no past history of asthma and no evidence of bronchial hyperresponsiveness (Table 1). Asthmatics and control subjects underwent fiberoptic bronchoscopy according to a standard protocol,lO and 8 to 10 endobronchial biopsies were performed through the bron-
Table I-Subject Characteristics and Cytoldne Releaae from Bronchial Epithelial Cells Over the Firat 48 H of Incubation Immunoreactive Cytokines,* nglmlll()& cellsl48 h Subject No.
FEV., %pred
Asthmatics 1 68.0 2 86.4 3 102.3 4 93.5 5 84.8 6 65.0 Control subjects 7 96.4 8 91.6 9 98.5 10 105.3 11 101.0 12 106.3
Atop~
+/-
0.03 0.22 1.12 0.39 0.03 0.49 >32 >32 >32 >32 >32 >32
+
+
+ +
GM-CSF
11.,6
ILB
2.50 0.85 0.61 1.52 1.91 1.28
1.85 0.81 0.54 0.96 1.12 0.41
2.65 1.60
Expression of the Potent Inflammatory Cytokines, GM-CSF; IL6, and 1L8, in Bronchial Epithelial Cells of Asthmatic Patients· Sabrina Mattoli, M.D., Ph.D.;t* Maurizio Marini, M.D.;* and Angelo Fasoli, M.D. t
C
ultured human bronchial epithelial cells produce cytokines with potent proinflammatory properties upon exposure to immunologic and nonimmunologic stimuli or during recovery from injury!-7 particularly GM-CSF and IL6. Because of this ability to release factors which affect the migration of granulocytes and lymphocytes and promote their local activation,2,3,6-9 airway epithelial cells can be viewed as cells that signal the inflammatory and immune system that penetration or disruption of the airway mucosa has occurred, and epithelial cell-derived cytokines may play an important role in the genesis and persistence of bronchial inflammation in some pathologic conditions, like asthma. Indeed, we have previously demonstrated that the airway secretions from patients with symptomatic asthma contain increased amounts ofGM-CSF and IL6, and these cytokines have been localized in the cytoplasm ofnonciliated bronchial epithelial cells by immunospecific labeling. 10 In addition, we have provided direct evidence of an upregulation of the expression of GM-CSF gene and protein in the bronchial epithelial cells of these patients. 6,7 The GM-CSF released from asthmatic bronchial epithelial cells could prolong the survival of eosinophils, make them hypodense, and promote *From the tSection of Lung Cellular and Molecular Biology, Department of Internal Medicine, University of Milan; and *Diagnostic Center for Respiratory Diseases, Milan, Italy. This study was supported by the Minister of University and Research, Italy. Reprint requests: Dr. Mattoli, Via C. Battisti 5, Tenli, Italy 05100
the release of mediators from those cells,6,7 thereby reproducing in vitro phenomena that are known to occur in asthma and that contribute to the pathogenesis of airway hyperresponsiveness. 11 In the present study, we have further evaluated the expression of cytokines with inflammatory properties in bronchial epithelial cells of asthmatic patients and tested the ability of corticosteroids and nedocromil sodium to downregulate their production. Six asthmatic patients (4 women, mean age ± SD: 31.2±9.8 years) and 6 healthy volunteers (3 women, mean age ± SD: 43.0 ± 15.5 years) completed the study. All subjects were nonsmokers and two in each group showed positive responses to skin prick testing with 12 common allergen extracts 12 (Table 1). The asthmatics were receiving treatment with inhaled bronchodilators alone, on demand, but their symptoms were not controlled, as they were elicited by nonspecific irritating stimuli, or required high doses of bronchodilators daily, or disturbed sleep. No patient had received corticosteroids in the last month. They had pulmonary function measurements and bronchoprovocation tests with methacholine 13 to define their nonspecific bronchial hyperresponsiveness. Their mean baseline FEV. (± SD) was 83.3 ± 14.5% of predicted value) and their geometric mean PCmM was 0.187 mglml (range from 0.04 to 0.844 mwml) (Table 1), the latter indicating that airway reactivity was moderately to severely increased in those patients. Control subjects had no past history of asthma and no evidence of bronchial hyperresponsiveness (Table 1). Asthmatics and control subjects underwent fiberoptic bronchoscopy according to a standard protocol,lO and 8 to 10 endobronchial biopsies were performed through the bron-
Table I-Subject Characteristics and Cytoldne Releaae from Bronchial Epithelial Cells Over the Firat 48 H of Incubation Immunoreactive Cytokines,* nglmlll()& cellsl48 h Subject No.
FEV., %pred
Asthmatics 1 68.0 2 86.4 3 102.3 4 93.5 5 84.8 6 65.0 Control subjects 7 96.4 8 91.6 9 98.5 10 105.3 11 101.0 12 106.3
Atop~
+/-
0.03 0.22 1.12 0.39 0.03 0.49 >32 >32 >32 >32 >32 >32
+
+
+ +
GM-CSF
11.,6
ILB
2.50 0.85 0.61 1.52 1.91 1.28
1.85 0.81 0.54 0.96 1.12 0.41
2.65 1.60
