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Diesel exhaust particulate induces airway hyperresponsiveness in a murine model: Essential role of GM-CSF  Ken Ohta, MD, PhD, Naomi Yamashita, MD, PhD,

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Presentation on theme: "Diesel exhaust particulate induces airway hyperresponsiveness in a murine model: Essential role of GM-CSF  Ken Ohta, MD, PhD, Naomi Yamashita, MD, PhD,"— Presentation transcript:

1 Diesel exhaust particulate induces airway hyperresponsiveness in a murine model: Essential role of GM-CSF  Ken Ohta, MD, PhD, Naomi Yamashita, MD, PhD, Makoto Tajima, MD, Takashi Miyasaka, MD, PhD, Junichi Nakano, MD, PhD, Mikio Nakajima, MD, Akira Ishii, MD, Tadashi Horiuchi, MD, PhD, Kenji Mano, MD, PhD, Terumasa Miyamoto, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 104, Issue 5, Pages (November 1999) DOI: /S (99) Copyright © 1999 Mosby, Inc. Terms and Conditions

2 Fig. 1 Airway hyperresponsiveness in A/J mice. Indicated concentrations of ACh were inhaled for 3 minutes. Raw (closed circles) and Cdyn (open triangles) were measured with use of whole-body plethysmographs as described in Methods. Arrowhead, Response obtained after injection of bronchial dilator terbutaline sulfate (BD, 0.5 μg). Data shown are means ± SEM for 5 mice. Asterisk, P < .05; two asterisks, P < .01. Journal of Allergy and Clinical Immunology  , DOI: ( /S (99) ) Copyright © 1999 Mosby, Inc. Terms and Conditions

3 Fig. 2 Airway hyperresponsiveness induced by DEP. After 2 weeks of intranasal DEP administration, airway responses to indicated concentrations of ACh were measured. Open triangles, DEP-treated A/J mice; closed triangles, untreated A/J mice; open circles, DEP-C57Bl/6 mice; closed circles, untreated C57Bl/6 mice. ACh (5 mg/mL)-evoked Raw is expressed as percentage of Raw observed before ACh inhalation (%Raw5). Data shown are means ± SEM of 5 independent experiments. Asterisk, P < .05. Journal of Allergy and Clinical Immunology  , DOI: ( /S (99) ) Copyright © 1999 Mosby, Inc. Terms and Conditions

4 Fig. 3 Duration of DEP-evoked airway hyperresponsiveness. Responses to ACh were measured 24 hours and 6 days after 2 weeks of intranasal DEP administration. ACh (5 mg/mL)-evoked Raw is expressed as percentage of Raw observed before ACh inhalation (%Raw5). Data shown are means ± SEM of 5 mice. Asterisk, P < .05. Journal of Allergy and Clinical Immunology  , DOI: ( /S (99) ) Copyright © 1999 Mosby, Inc. Terms and Conditions

5 Fig. 4 Effects of muscarinic antagonists on DEP-induced increases in airway responsiveness. M 3 muscarinic receptor antagonist 4-DAMP (0.2 μg/kg) was administered intraperitoneally 15 minutes before ACh inhalation. ACh (5 mg/mL)-evoked Raw is expressed as percentage of Raw observed before ACh inhalation (%Raw5). Data shown are means ± SEM of 7 mice. Two asterisks, P < .01. Journal of Allergy and Clinical Immunology  , DOI: ( /S (99) ) Copyright © 1999 Mosby, Inc. Terms and Conditions

6 Fig. 5 Effects of anti-GM-CSF antibody on DEP-evoked increases in airway responsiveness to ACh. Anti-GM-CSF or anti-IL-4 neutralizing antibody or goat IgG (sham control) was administered intranasally for 2 weeks immediately before each DEP treatment. Data are expressed as percentages of Raw measured before intranasal administration of 5 mg/mL ACh (%Raw5). Data shown are means ± SEM of 8 mice. Asterisk, P < .05; two asterisks, P < .01. Journal of Allergy and Clinical Immunology  , DOI: ( /S (99) ) Copyright © 1999 Mosby, Inc. Terms and Conditions

7 Fig. 6 Analysis of bronchoalveolar lavage fluid (BALF). Lungs were subjected to lavage through intubation tubes with 5 mL of saline solution (0.9 mol/L sodium chloride), and the lavage fluid was collected. Cells present in BALF were pelleted, resuspended in 1 mL of saline solution, and placed on glass slides, where they were counted and fixed with Cytospin. The slides were then stained with Diff Quik, and cell differentiation was assessed microscopically. Data shown are means ± SEM of 8 mice. Two asterisks, P < .01. Journal of Allergy and Clinical Immunology  , DOI: ( /S (99) ) Copyright © 1999 Mosby, Inc. Terms and Conditions

8 Fig. 7 Effect of anti-GM-CSF neutralizing antibody on the histologic characteristics of lung tissue. Mice were treated with anti-GM-CSF antibody and DEP as described in the legend to Fig 4. Tissue specimens from (A) untreated control mouse, (B) DEP-treated mouse, (C) mouse treated with both anti-GM-CSF and DEP, and (D) mouse treated with both goat IgG and DEP. Journal of Allergy and Clinical Immunology  , DOI: ( /S (99) ) Copyright © 1999 Mosby, Inc. Terms and Conditions

9 Fig. 8 PCR analysis of expression of mRNAs encoding GM-CSF, IL-4, IL-5, and TNF-α. For control (lanes 1, 3, 5, 7, and 9) and DEP-treated (lanes 2, 4, 6, 8, and 10) mice, respectively, lanes 1 and 2 contain mRNAs encoding β-actin; lanes 3 and 4, GM-CSF; lanes 5 and 6, IL-4; lanes 7 and 8, IL-5; and lanes 9 and 10, TNF-α. DNA size standards are shown in the first lane on the left.. Journal of Allergy and Clinical Immunology  , DOI: ( /S (99) ) Copyright © 1999 Mosby, Inc. Terms and Conditions

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