Eosinophils generate brominating oxidants in allergen-induced asthma

W Wu

M Samoszuk

S Comhair

M Thomassen

^{Department of Cell Biology, Cleveland Clinic Foundation, Cleveland, Ohio, USA}^{Chemistry Department, Cleveland State University, Cleveland, Ohio, USA}^{Quest Diagnostics Inc., San Juan Capistrano, California, USA}^{Department of Cancer Biology,}^{Department of Pulmonary and Critical Care Medicine,}^{Department of Anatomic Pathology, and}^{Department of Cardiology, Cleveland Clinic Foundation, Cleveland, Ohio, USA}

Address correspondence to: Stanley L. Hazen, Cleveland Clinic Foundation, Lerner Research Institute, Department of Cell Biology, 9500 Euclid Avenue, NC-10, Cleveland, Ohio 44195, USA. Phone: (216) 445-9763; Fax: (216) 444-9404; E-mail: lmth.snezah/ip/ir/gro.fcc.renrel.www//:ptth :LRU ;gro.fcc@snezah.

Received 2000 Feb 24; Accepted 2000 Mar 27.

Abstract

Eosinophils promote tissue injury and contribute to the pathogenesis of allergen-triggered diseases like asthma, but the chemical basis of damage to eosinophil targets is unknown. We now demonstrate that eosinophil activation in vivo results in oxidative damage of proteins through bromination of tyrosine residues, a heretofore unrecognized pathway for covalent modification of biologic targets in human tissues. Mass spectrometric studies demonstrated that 3-bromotyrosine serves as a specific “molecular fingerprint” for proteins modified through the eosinophil peroxidase-H₂O₂ system in the presence of plasma levels of halides. We applied a localized allergen challenge to model the effects of eosinophils and brominating oxidants in human lung injury. Endobronchial biopsy specimens from allergen-challenged lung segments of asthmatic, but not healthy control, subjects demonstrated significant enrichments in eosinophils and eosinophil peroxidase. Baseline levels of 3-bromotyrosine in bronchoalveolar lavage (BAL) proteins from mildly allergic asthmatic individuals were modestly but not statistically significantly elevated over those in control subjects. After exposure to segmental allergen challenge, lung segments of asthmatics, but not healthy control subjects, exhibited a >10-fold increase in BAL 3-bromotyrosine content, but only two- to threefold increases in 3-chlorotyrosine, a specific oxidation product formed by neutrophil- and monocyte-derived myeloperoxidase. These results identify reactive brominating species produced by eosinophils as a distinct class of oxidants formed in vivo. They also reveal eosinophil peroxidase as a potential therapeutic target for allergen-triggered inflammatory tissue injury in humans.

Abstract

Acknowledgments

Mass spectrometry experiments were performed at the Lerner Research Institute Mass Spectrometry Facility. This work was supported in part by the American Heart Association and by the National Institutes of Health (grants HL-61878 and HL-62526).

Acknowledgments

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