title: An airway epithelial iNOS-DUOX2-thyroid peroxidase metabolome drives Th1/Th2 nitrative stress in human severe asthma.
abstract: Severe refractory asthma is associated with enhanced nitrative stress. To determine the mechanisms for high nitrative stress in human severe asthma (SA), 3-nitrotyrosine (3NT) was compared with Th1 and Th2 cytokine expression. In SA, high 3NT levels were associated with high interferon (IFN)-γ and low interleukin (IL)-13 expression, both of which have been reported to increase inducible nitric oxide synthase (iNOS) in human airway epithelial cells (HAECs). We found that IL-13 and IFN-γ synergistically enhanced iNOS, nitrite, and 3NT, corresponding with increased H(2)O(2). Catalase inhibited whereas superoxide dismutase enhanced 3NT formation, supporting a critical role for H(2)O(2), but not peroxynitrite, in 3NT generation. Dual oxidase-2 (DUOX2), central to H(2)O(2) formation, was also synergistically induced by IL-13 and IFN-γ. The catalysis of nitrite and H(2)O(2) to nitrogen dioxide radical (NO(2)(•)) requires an endogenous peroxidase in this epithelial cell system. Thyroid peroxidase (TPO) was identified by microarray analysis ex vivo as a gene distinguishing HAEC of SA from controls. IFN-γ induced TPO in HAEC and small interfering RNA knockdown decreased nitrated tyrosine residues. Ex vivo, DUOX2, TPO, and iNOS were higher in SA and correlated with 3NT. Thus, a novel iNOS-DUOX2-TPO-NO(2)(•) metabolome drives nitrative stress in HAEC and likely in SA.
authors: 1] University of Pittsburgh Asthma Institute at UPMC and the University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA [2] Department of Pediatrics, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
authors: N
authors: N
authors: Voraphani
authors: Division of Pulmonary, Allergy, and Critical Care Medicine, Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors: M T
authors: MT
authors: Gladwin
authors: 1] University of Pittsburgh Asthma Institute at UPMC and the University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA [2] Division of Pulmonary, Allergy, and Critical Care Medicine, Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors: A U
authors: AU
authors: Contreras
authors: Division of Pulmonary, Allergy, and Critical Care Medicine, Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors: N
authors: N
authors: Kaminski
authors: Division of Pulmonary, Allergy, and Critical Care Medicine, Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors: J R
authors: JR
authors: Tedrow
authors: Division of Pulmonary, Allergy, and Critical Care Medicine, Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors: J
authors: J
authors: Milosevic
authors: Center for Genomics and Personalized Medicine Research, Wake Forest School of Medicine, Winston-Salem, North Carolina, USA.
authors: E R
authors: ER
authors: Bleecker
authors: Center for Genomics and Personalized Medicine Research, Wake Forest School of Medicine, Winston-Salem, North Carolina, USA.
authors: D A
authors: DA
authors: Meyers
authors: 1] University of Pittsburgh Asthma Institute at UPMC and the University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA [2] Division of Pulmonary, Allergy, and Critical Care Medicine, Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors: A
authors: A
authors: Ray
authors: 1] University of Pittsburgh Asthma Institute at UPMC and the University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA [2] Division of Pulmonary, Allergy, and Critical Care Medicine, Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors: P
authors: P
authors: Ray
authors: Department of Pathobiology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio, USA.
authors: S C
authors: SC
authors: Erzurum
authors: Division of Allergy and Clinical Immunology, Department of Medicine, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin, USA.
authors: W W
authors: WW
authors: Busse
authors: 1] University of Pittsburgh Asthma Institute at UPMC and the University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA [2] Division of Pulmonary, Allergy, and Critical Care Medicine, Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors: J
authors: J
authors: Zhao
authors: 1] University of Pittsburgh Asthma Institute at UPMC and the University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA [2] Division of Pulmonary, Allergy, and Critical Care Medicine, Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors: J B
authors: JB
authors: Trudeau
authors: 1] University of Pittsburgh Asthma Institute at UPMC and the University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA [2] Division of Pulmonary, Allergy, and Critical Care Medicine, Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors: S E
authors: SE
authors: Wenzel
mesh_terms: D000328:Adult; D001249:Asthma / Q000201:enzymology* / Q000276:immunology / Q000503:physiopathology*; D005260:Female; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D018793:Interleukin-13 / Q000494:pharmacology; D007453:Iodide Peroxidase / Q000378:metabolism; D008297:Male; D055442:Metabolome; D046228:Microarray Analysis; D052247:Nitric Oxide Synthase Type II / Q000276:immunology*; D012137:Respiratory System / Q000201:enzymology / Q000503:physiopathology; D012720:Severity of Illness Index; D013312:Stress, Physiological / Q000187:drug effects; D018417:Th1 Cells / Q000187:drug effects / Q000276:immunology*; D018418:Th2 Cells / Q000187:drug effects / Q000276:immunology*; D055815:Young Adult
chemical_list: D018793:Interleukin-13; D007371:Interferon-gamma; D007453:Iodide Peroxidase; D052247:Nitric Oxide Synthase Type II