Nitrotyrosine formation in the airways and lung parenchyma of patients with asthma,☆☆,

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Abstract

Background: Recent evidence has shown that nitric oxide (NO) levels are increased in asthmatic airways. Although the role of NO in asthma is unknown, reactive metabolites of NO may lead to nitrotyrosine formation and promote airway dysfunction. Objective: The aim of this study was to determine whether nitrotyrosine, as a marker of nitrating species, could be found in the airways and lung parenchyma of subjects with asthma who died of status asthmaticus or other nonrespiratory causes. Methods: Lung tissue specimens were obtained from 5 patients who died of status asthmaticus, 2 asthmatic patients who died of nonrespiratory causes, and 6 nonasthmatic control subjects who died of nonrespiratory causes. Lung sections were stained for immunofluorescence with use of an antinitrotyrosine antibody, followed by a indiocarbocyanine (Cy5, Jackson Immunochemicals, Westgrove, Pa)–conjugated secondary antibody. Results: Nonasthmatic lungs showed little or no nitrotyrosine staining, whereas asthmatic lungs demonstrated significantly more staining of nitrotyrosine residues distributed in both the airways and lung parenchyma. Conclusion: This study demonstrates the presence of nitrotyrosine, and hence evidence of formation of nitrating species, in the airways and lung parenchyma of patients with asthma who died of status asthmaticus or other nonrespiratory causes. This finding supports the concept that widespread airway and parenchymal inflammation occurs in asthma, and, more specifically, that NO and its reactive metabolites may play a pathophysiologic role in asthma. (J Allergy Clin Immunol 1999;104:747-54.)

Section snippets

Tissue specimens and routine histologic examination

Lung tissue specimens were obtained at autopsies performed at Fletcher Allen Health Care, the teaching hospital of the University of Vermont College of Medicine, and from the Sir Charles Gairdner Hospital of Nedlands, Australia. Clinical data on the subjects whose lung tissue was analyzed were collected from review of coroner and hospital records. The protocol was approved by the Committees on Human Research in the Medical Sciences at the University of Vermont and the Sir Charles Gairdner

RESULTS

The clinical characteristics of the subjects involved in the study are shown in Table I. There were 7 asthmatic and 6 nonasthmatic subjects. Of the 7 asthmatic patients, 5 had died of status asthmaticus and 2 had died from other nonrespiratory causes. These latter 2 subjects were both smokers and atopic and were taking inhaled β-agonists regularly but were not receiving corticosteroids nor were ever hospitalized for asthma. The 5 subjects with fatal asthma were all nonsmokers and atopic and

DISCUSSION

This study has demonstrated the presence of nitrotyrosine and hence evidence of formation of reactive nitrogen species in the airways and lung parenchyma of patients with asthma who died of status asthmaticus or other nonrespiratory causes.

Before the implications of these findings are considered, a few technical considerations are important. First, we assumed that the presence of immunoreactivity against nitrotyrosine was evidence for the formation of reactive nitrogen species. Nitrotyrosine

Acknowledgements

We thank Marilyn Wadsworth of the University of Vermont Cell Imaging Facility for technical assistance, Gary Nelson of Medical Illustration at the University of Vermont College of Medicine for assistance with the schematic diagram, and Charles G. Irvin, PhD, and Gerald S. Davis, MD, of the University of Vermont College of Medicine for their review of the manuscript.

References (24)

  • S Kharitonov et al.

    Increased nitric oxide in exhaled air of asthmatic patients

    Lancet

    (1994)
  • WJ Calhoun et al.

    Enhanced reactive oxygen species metabolism of airspace cells and airway inflammation follow antigen challenge in human asthma

    J Allergy Clin Immunol

    (1990)
  • K Alving et al.

    Increased amount of nitric oxide in exhaled air of asthmatics

    Eur Respir J

    (1993)
  • S Kharitonov et al.

    Allergen-induced late asthmatic reactions are associated with elevation of exhaled nitric oxide

    Am J Respir Crit Care Med

    (1995)
  • A Jatakanon et al.

    Correlation between exhaled nitric oxide, sputum eosinophils and methacholine responsiveness in patients with mild asthma

    Thorax

    (1998)
  • M Hogman et al.

    Inhalation of nitric oxide modulates adult human bronchial tone

    Am Rev Respir Dis

    (1993)
  • N Kageyama et al.

    Role of endogenous nitric oxide in airway microvascular leakage induced by inflammatory mediators

    Eur Respir J

    (1997)
  • T Flak et al.

    Autotoxicity of nitric oxide in airway disease

    Am J Respir Crit Care Med

    (1996)
  • J Beckman et al.

    Apparent hydroxyl radical production by peroxynitrite: implications for endothelial injury from nitric oxide and superoxide

    Proc Natl Acad Sci U S A

    (1990)
  • I Haddad et al.

    Quantitation of nitrotyrosine levels in lung sections of patients and animals with acute lung injury

    J Clin Invest

    (1994)
  • J Eiserich et al.

    Formation of nitric oxide–derived inflammatory oxidants by myeloperoxidase in neutrophils

    Nature

    (1998)
  • J Beckman et al.

    Nitric oxide, superoxide, and peroxynitrite: the good, the bad, and the ugly

    Am J Physiol

    (1996)
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    Supported by National Institutes of Health grant No. K08 HL03517 and the American Lung Association of Vermont (D. A. K.) and Parker B. Francis Foundation and National Institute for Occupational Safety and Health grant No. R03 OH03457 (Y. J. H.).

    ☆☆

    Reprint requests: David A. Kaminsky, MD, Pulmonary Disease and Critical Care Medicine, University of Vermont College of Medicine, Given C-317, Burlington, VT 05405.

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