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RS Virus-Induced Inflammation and the Intracellular Glutathione Redox State in Cultured Human Airway Epithelial Cells

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Abstract

There is ample evidence that asthma is mediated by oxidative stress and that viral infection, which is associated with asthma onset and exacerbation in infants, acts as one type of oxidative stress. The goal of this study was to determine whether respiratory syncytial virus (RSV) induces oxidative stress in cultured A549 human airway epithelial cells and normal human bronchial epithelial cells (NHBE), and whether such RSV-induced oxidative stress can induce airway inflammation. To evaluate the direct effect of RSV infection as an oxidative stressor, the intracellular levels of reduced glutathione (GSH) or oxidized glutathione (GSSG) were measured. Their ratio (GSH/GSSG) was calculated to indicate intracellular oxidation–reduction (redox) status in A549 and NHBE. To evaluate the extent to which glutathione redox regulation affected cytokine/chemokine production, the effect of pretreatment with a reductive agent, glutathione monoethyl ester (GSH-OEt) and RSV-specific monoclonal antibody was thus studied. RSV acted as a potent oxidative stressor on the intracellular glutathione redox state in human airway epithelial cells, activating signals to increase the production of cytokine/chemokine. Pretreatment with GSH-OEt significantly suppressed RSV-induced time-dependent changes in the intracellular redox state, and also suppressed RSV-induced up-regulation of epithelial cell-derived IL-8, IL-6 and eotaxin production, as well as RSV-specific monoclonal antibody. RSV-induced oxidative stress is likely to contribute to the perpetuation and amplification of the inflammatory response. Therapeutic intervention against oxidative stress might therefore be beneficial as adjunctive therapies for respiratory disorders that are caused by an RSV infection.

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Abbreviations

BPE:

Bovine pituitary extract

DTNB:

5-5,-dithiobis-2-nitrobenzoic acid

DTT:

Dithiothreitol

EDTA:

Ethylenediamine tetraacetate

hEGF:

Human recombinant epidermal growth factor

FCS:

Fetal calf serum

GSH:

Reduced glutathione

GSH-OEt:

Glutathione monoethyl ester

GSSG:

Oxidized GSH

H2O2 :

Superoxide radicals and hydrogen peroxide

IRF-3:

Interferon regulatory factor 3

MCP:

Monocyte chemotactic protein

β-NADPH:

Nicotinamide adenine dinucleotide phosphate

NF-κB:

Nuclear factor-κB

NHBE:

Normal human bronchial epithelial cells

Redox:

Reduction–oxidation

RSV:

Respiratory syncytial virus

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Acknowledgments

We thank Junying Wang, M.D, Mitsuyoshi Utsugi M.D., Kunio Dobashi M.D., Masatomo Mori M.D., Ms. Tomoko Endo and Ms. Chinori Iijima (Gunma University, Maebashi, Japan) for their valuable general scientific and technical support.

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  1. Department of Pediatrics, Gunma University Graduate School of Medicine, 3-39-15 Showa-machi, Maebashi, Gunma, 371-8511, Japan

    Hiroyuki Mochizuki, Makoto Todokoro & Hirokazu Arakawa

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  1. Hiroyuki Mochizuki
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Correspondence to Hiroyuki Mochizuki.

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Mochizuki, H., Todokoro, M. & Arakawa, H. RS Virus-Induced Inflammation and the Intracellular Glutathione Redox State in Cultured Human Airway Epithelial Cells. Inflammation 32, 252–264 (2009). https://doi.org/10.1007/s10753-009-9128-0

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