Role of the eosinophil in protein oxidation in asthma: Possible effects on proteinase/antiproteinase balance

Background: Many of the leukocytes which migrate into the tissue following allergen challenge can undergo a respiratory burst producing reactive oxygen species causing tissue damage and distorting proteinase/antiproteinase balance. The reactive oxygen species have extremely short half lives and so cannot be measured in vivo, but the protein carbonyl residues which result from protein oxidation can be measured in biological fluids. Methods: We examined protein oxidation in bronchoalveolar lavage after allergen challenge in 12 patients with atopic asthma by measuring protein carbonyl residues using a sensitive Western blotting technique. Results: We found that the median level of protein carbonyls per molecule of protein rose from 0.09 in bronchoalveolar lavage (BAL) obtained 18 h after saline challenge to 0.23 in BAL taken 10 min after segmental allergen challenge, reaching a median of 0.82 in samples obtained 18 h later (p<0.01 compared with saline control and the earlier time point). The number of protein carbonyl residues correlated strongly with the number of eosinophils recovered in the BAL (rho = 0.574, p<0.05) but not with the number of neutrophils (rho = 0.228, p = NS) or macrophages (rho = 0.178, p = NS). Western blot ting showed that the majority of the modified protein comigrated with authentic alpha 1-antitrypsin at 53 kD. In contrast, BAL samples from patients with chronic obstructive pulmonary disease, which is characterized by an influx of neutrophils, showed that the main oxidized protein colocalized with human serum albumin, the predominant protein in the BAL. Conclusion: Our results suggest that the recruitment and activation of eosinophils accounts for much of the protein oxidation found in the BAL following allergen challenge. The main oxidized protein appears to be alpha 1-antitrypsin, a key antiproteinase in the airways. Inactivation of alpha 1-antitrypsin by oxidation may distort the proteinase/antiproteinase balance leaving the lung vunerable to proteolytic damage. Intriguingly, we have evidence to suggest that other airways diseases, characterized by recruitment of other inflammatory cells, may result in the oxidation of alternative targets.