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Inhibiting pollen reduced nicotinamide adenine dinucleotide phosphate oxidase–induced signal by intrapulmonary administration of antioxidants blocks allergic.

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Presentation on theme: "Inhibiting pollen reduced nicotinamide adenine dinucleotide phosphate oxidase–induced signal by intrapulmonary administration of antioxidants blocks allergic."— Presentation transcript:

1 Inhibiting pollen reduced nicotinamide adenine dinucleotide phosphate oxidase–induced signal by intrapulmonary administration of antioxidants blocks allergic airway inflammation  Nilesh Dharajiya, MD, Barun K. Choudhury, PhD, Attila Bacsi, PhD, Istvan Boldogh, PhD, Rafeul Alam, MD, PhD, Sanjiv Sur, MD  Journal of Allergy and Clinical Immunology  Volume 119, Issue 3, Pages (March 2007) DOI: /j.jaci Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Antioxidants inhibit RWE mediated oxidative stress in vitro. ID50 of individual (A) and combination (B) of antioxidants. GSH, Glutathione; Toc, tocopherol. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Effect of antioxidants on RWE-induced intracellular ROS and 4-HNE protein adduct formation. A, Effect of increasing concentration of antioxidants on RWE-induced intracellular ROS in A549 cells. B, Effect of NAC on RWE-induced 4-HNE adduct formation in lungs. Each lane represents a different mouse. GO, Glucose oxidase. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Effect of AA+NAC on RWE-induced mucin production. A, RWE-sensitized BALB/c mice were challenged with PBS, RWE, or RWE+AA+NAC and killed at 72 hours. The PAS-stained lung sections show mucin content of airway epithelial cells. Arrow shows PAS-positive mucin globules. Magnification ×200. B, Morphometric quantification of mucin in airway epithelial cells. Results are means ± SEMs (n = 3-6 mice per group). C, Measurement of Clca3, IL-4, and IL-13 transcripts in the lungs by real-time quantitative PCR. ∗∗P < .01; ∗∗∗P < .005. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Scavenging pollen NADPH oxidase–generated ROS inhibits allergic lung inflammation. Total inflammatory cells (A) and eosinophils (B) in BAL fluids of RWE-challenged mice. Results are means ± SEMs (n = 7-9 mice per group). ∗∗∗P < C, Effect of antioxidants on recruitment of eosinophils in peribronchial (br) and perivascular (v) regions (arrows). Magnification ×100. TOC, Tocopherol. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 Effect of antioxidants on X+XO-mediated augmentation of allergic airway inflammation. Effect of antioxidants on X+XO-augmented total inflammatory cell (A) and eosinophil (B) recruitment in BAL fluids. Results are means ± SEMs (n = 7-9 mice per group). ∗∗P < .01; ∗∗∗P < .001. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 Scavenging ROS generated by RWE pollen NADPH oxidase and not by inflammatory cells inhibits allergic airway inflammation. A, Kinetics of neutrophil recruitment in the BAL fluid. RWE-sensitized mice were challenged with either PBS or RWE and killed kinetically, and BAL fluid was examined for cellular content. B, AA+NAC increased total antioxidant potential in the airways for 90 minutes. X-axis represents time after administration of antioxidants. Results are means ± SEMs (n = 3-6 mice per group). C, Analysis of eosinophil recruitment in the airways after administration of antioxidants at various time points. Results are means ± SEMs (n = 7-9 mice per group). ∗P < .05; ∗∗∗∗P < D, BALB/c mice were sensitized with OVA and challenged with nebulized OVA. Mice were treated with either PBS or AA+NAC immediately before and just after nebulized OVA challenge. NS, Not significant. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig 7 Antioxidants inhibit second RWE challenge induced augmentation of allergic airway inflammation. A, AA+NAC prevents further increase in airway inflammation induced by second RWE challenge performed 72 hours after initial challenge. RWE-sensitized mice were challenged with PBS, RWE, or RWE+AA+NAC. A second similar challenge was performed 72 hours after the initial challenge; mice were killed 72 hours after last challenge, and BAL fluid was examined for cellular content. B, AA+NAC effectively inhibits RWE-induced airway inflammation during resolution phase. Mice were sensitized and challenged as described except the second challenge was performed 10 days after the first RWE challenge. ∗P < .05; ∗∗P < .01. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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