Mechanisms of allergy and clinical immunology
Leukocyte nicotinamide adenine dinucleotide phosphate-reduced oxidase is required for isocyanate-induced lung inflammation

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Background

Isocyanates are low-molecular-weight compounds noted for inducing occupational and environmental asthma. Isocyanate-induced lung disease, an oxidant stress-dependent pulmonary inflammation, is the leading cause of occupational asthma.

Objectives

To address the role of leukocyte-produced oxidants in airway inflammation induced by toluene diisocyanate (TDI), and to elucidate the role of leukocyte nicotinamide adenine dinucleotide phosphate-reduced (NADPH) oxidase in pathogenesis by TDI.

Methods

Wild-type mice and NADPH oxidase–deficient mice (neutrophil cytosolic factor 1 mutant, Ncf1–/–) were intranasally injected, challenged with inhalatory TDI, and then investigated for lung inflammation.

Results

Cell infiltration in lung tissue and leukocytes in bronchoalveolar lavage, airway reactivity to a methacholine challenge, and TDI-induced inflammatory cytokine expression and nuclear factor activation in the lung tissue were all markedly lower in Ncf1–/– mice. Wild-type mice treated with blocking antibodies against CD4 and IL-17 showed markedly lower TDI-induced airway hyperresponsiveness.

Conclusion

Leukocyte NADPH oxidase is an essential regulator in TDI-induced airway inflammation through redox modification of immune responses.

Section snippets

Animals

Female, B6 (Cg)-Ncf1<m1J>/J mice (stock number 004742) were obtained from Jackson Laboratory (Bar Harbor, Me). Female wild-type C57BL/6J mice were from the Laboratory Animal Center at National Cheng Kung University. The experimental protocol was approved by the Institutional Animal Care and Use Committee of National Cheng Kung University.

Reagents

Toluene diisocyanate isomers were purchased from Merck (Darmstadt, Germany). Ovalbumin (OVA), N-acetyl-L-cysteine (NAC), 5, 5′-dithiobis-(2-nitrobenzoic acid)

TDI exposure and leukocyte NADPH oxidase activity induce oxidant stress in the lung

We first examined whether TDI exposure stimulates the generation of ROS in the airway. Wild-type mice were sensitized, then challenged with TDI and killed 24 hours after the last challenge. The bronchoalveolar lavage (BAL) cells were collected and incubated with H2DCFDA to detect the production of intracellular H2O2. We found that H2O2 production significantly increased in BAL cells from wild-type mice treated with TDI compared with equal numbers of BAL cells from the mice without TDI treatment

Discussion

By inducing lung inflammation with TDI, a chemical that causes oxidant stress in a mouse model with or without deficient leukocyte NADPH oxidase, we showed that leukocyte NAPDH oxidase plays a crucial role in TDI-induced lung damage.

Although the evidence for the role of oxidant stress in lung inflammation has been accumulating in the past decade, the relationship between exposure to different allergens and redox-mediated leukocyte activation remains elusive.29, 30 Airway inflammation in asthma

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    Supported by grants from the National Science Council, Taiwan. C.-C.S. was supported by grants from the National Science Council and National Health Research Institute, Taiwan.

    Disclosure of potential conflict of interest: The authors have declared that they have no conflict of interest.

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    Copyright © 2011 American Academy of Allergy, Asthma & Immunology. Published by Mosby, Inc. All rights reserved.