An oxidant-antioxidant imbalance with damaging consequences for the bronchial epithelium has been hypothesized in the airways of patients with cystic fibrosis (CF). It is based on the assumption that neutrophils entering the lumen of the infected airways undergo activation and release toxic oxygen metabolites and myeloperoxidase (MPO), an enzyme which transforms hydrogen peroxide (H2O2) into highly toxic oxygen metabolites. Our aims were to substantiate this hypothesis. H2O2 levels were measured in breath condensates of 63 CF patients and 51 normal subjects. In CF sputum samples, activities and concentrations of MPO and catalase (CAT) were determined. MPO/H2O2-mediated cytotoxicity of CF sputum was measured in cell culture assays. H2O2 levels were similar in CF patients and normal subjects (mean +/-SD) 0.97 +/- 0.69 versus 1.11+/-0.78 microM; p=0.427). Concentrations and activities of CAT (0.31+/-0.18 microM; 105+/-69 units) and MPO (5.93+/-4.8 microM; 87.8+/-75 units) were detectable in 38 CF sputa. Addition of H2O2 to in vitro cells preincubated with CF sputum did not induce cytotoxicity even when CAT was removed from sputum. Sputum MPO together with H2O2 did not inactivate alpha-proteinase inhibitor. Preincubation of MPO with sulphated glycoconjugates or deoxyribonucleic acid (DNA) totally inhibited its cytotoxic effect. In conclusion, catalase, sulphated glycoconjugates and deoxyribonucleic acid may prevent myeolperoxidase-mediated oxygen radical generation in cystic fibrosis sputum.