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Weiguo Chen, MD, PhD, Yasuhiro Tabata, MD, PhD, Aaron M

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Presentation on theme: "Weiguo Chen, MD, PhD, Yasuhiro Tabata, MD, PhD, Aaron M"— Presentation transcript:

1 Matrix metalloproteinase 8 contributes to solubilization of IL-13 receptor α2 in vivo 
Weiguo Chen, MD, PhD, Yasuhiro Tabata, MD, PhD, Aaron M. Gibson, BS, Michael O. Daines, MD, Manoj R. Warrier, MD, Marsha Wills-Karp, PhD, Gurjit K. Khurana Hershey, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 122, Issue 3, Pages (September 2008) DOI: /j.jaci Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Levels of soluble IL-13Rα2 in serum and BALF and the expression of MMPs in lungs from PBS- or HDM-treated FVB/N mice. A and B, Soluble IL-13Rα2 in serum and BALF. Data were shown as means ± SD (n = 4-5). ∗P < .05. The result is representative of 3 experiments. C, RNase protection assay of the RNA from the lungs of PBS- or HDM-treated mice (n = 4). TIMP, Tissue inhibitor of metalloproteinases; GAPDH, glyceraldehyde-3-phosphate dehydrogenase. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Acellular cleavage of GST–IL-13Rα2–FLAG fusion protein by MMPs. A, Cleavage of GST–mIL-13Rα2–FLAG. B and C, Cleavage of GST or GST–mIL-13Rα2–FLAG by MMP-8. D and E, Cleavage of GST–mIL-13Rα2–FLAG or human collagen type I by MMP-8 in the presence or absence of an MMP-8 inhibitor or mock MMP-8 inhibitor control (5, 50, or 500 nmol/L). F, Cleavage of GST–hIL-13Rα2–FLAG. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Measurement of soluble IL-13Rα2 in the culture medium and on the cell surface after MMP-8 treatment. Soluble IL-13Rα2 levels are shown in panels A and B. IL-13Rα2 surface expression is shown as mean channel fluorescence (MCF) in panels C and D. Data are shown as means ± SD (n = 3-5). ∗P < .05. The result is representative of 3 experiments. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 IL-13 binding of soluble IL-13Rα2 in culture medium from MMP-8–treated cells. The result is representative of 3 experiments. A, MMP-8 treatment of untransfected or mIL-13Rα2–transfected WEHI cells. The data are shown as means ± SD (n = 5). ∗P < .05. B, Western blot analysis of MMP-8 treatment of murine GST–IL-13Rα2–FLAG fusion protein for the indicated time ( hours). U, Untreated. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 AHR, airway inflammation, BALF soluble IL-13Rα2 and its IL-13 binding activity, and BALF IL-13 of PBS- or HDM-treated wild-type and MMP-8–deficient mice. A, APTI (n = 6-8). B, BAL cell counting (n = 6-8). C, Soluble IL-13Rα2 (n = 6-8). D, IL-13 binding activity of soluble IL-13Rα2 (n = 12-17, pooled from 2 experiments). E, Free and IL-13Rα2–bound IL-13 (n = 5). WT, Wild-type mice; KO, MMP-8–deficient mice. The data are shown as means ± SD. ∗P < .05. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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