Transforming growth factor β1 increases fibronectin deposition through integrin receptor α5β1 on human airway smooth muscle Lyn M. Moir, PhD, Janette.

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Presentation transcript:

Transforming growth factor β1 increases fibronectin deposition through integrin receptor α5β1 on human airway smooth muscle Lyn M. Moir, PhD, Janette K. Burgess, PhD, Judith L. Black, MBBS, PhD Journal of Allergy and Clinical Immunology Volume 121, Issue 4, Pages 1034-1039.e4 (April 2008) DOI: 10.1016/j.jaci.2007.12.1159 Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Integrin-dependent FBS-induced proliferation of nonasthmatic (A; n = 14) and asthmatic (B; n = 9) ASM cells. Proliferation was assessed after culture for 6 days in the presence or absence of fb-mAbs to β1 either alone or in combination with anti-α5. Data are means ± SEMs of triplicate values from individual experiments. ∗P < .05 compared with IgG control. #P < .05 compared with 0.1% FBS. Journal of Allergy and Clinical Immunology 2008 121, 1034-1039.e4DOI: (10.1016/j.jaci.2007.12.1159) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Cell-surface expression of α5 on ASM cells from nonasthmatic (A and C) and asthmatic (B and D) subjects. Representative flow cytometric histograms of nonasthmatic (Fig 2, A) and asthmatic (Fig 2, B) ASM cells showing α5 expression after stimulation in the presence (gray line) or absence (black line) of TGF-β (1 ng/mL) for 48 hours are shown. Inset panels show isotype controls. ∗P ≤ .05 compared with 0.1% ITS. GM, Geometric mean. Journal of Allergy and Clinical Immunology 2008 121, 1034-1039.e4DOI: (10.1016/j.jaci.2007.12.1159) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Production of α5(A) and β1(B) mRNA by nonasthmatic (n = 6-9) and asthmatic (n = 5) ASM cells in response to TGF-β (1 ng/mL; black bars) at 24 hours. Results are expressed as a fold increase above the time-matched control (0.1% ITS; white bars). ∗P ≤ .05 compared with control. Journal of Allergy and Clinical Immunology 2008 121, 1034-1039.e4DOI: (10.1016/j.jaci.2007.12.1159) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Production of α5 mRNA by nonasthmatic (white bars) and asthmatic (black bars) ASM cells after stimulation with TGF-β (1 ng/mL) in the presence or absence of the ERK inhibitor U0126 (A) and the PI3K inhibitors LY294002 (B) or wortmannin (C) at 24 hours. Results are expressed relative to TGF-β–induced expression (gray bars). ∗P < .05 compared with TGF-β. Journal of Allergy and Clinical Immunology 2008 121, 1034-1039.e4DOI: (10.1016/j.jaci.2007.12.1159) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 The effect of α5β1 function-blocking antibodies on fibronectin deposition. Fibronectin matrix visualized by means of immunofluorescence (red staining) in untreated (0.1% ITS) cells (A), TGF-β–stimulated cells (B), TGF-β–stimulated cells exposed to isotype control antibodies (C) and TGF-β–stimulated cells exposed to α5 and β1 fb-mAbs (D) is shown. A representative experiment from 1 nonasthmatic subject is shown. Journal of Allergy and Clinical Immunology 2008 121, 1034-1039.e4DOI: (10.1016/j.jaci.2007.12.1159) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

The effect of TGF-β (1 ng/mL) stimulation on the proliferation of ASM cells from nonasthmatic (n = 6; white bars) and asthmatic (n = 5; black bars) subjects. Proliferation was assessed after stimulation for 24 hours in the presence or absence of TGF-β (1 ng/mL). Results are expressed as a percentage of the time-matched control (0.1% ITS). Journal of Allergy and Clinical Immunology 2008 121, 1034-1039.e4DOI: (10.1016/j.jaci.2007.12.1159) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Representative flow cytometric plots showing the number of integrin α5–positive ASM cells from nonasthmatic (B) and asthmatic (D) subjects compared with an isotype-matched IgG antibody (nonasthmatic [A] and asthmatic [C]). Journal of Allergy and Clinical Immunology 2008 121, 1034-1039.e4DOI: (10.1016/j.jaci.2007.12.1159) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions