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Increased IL-13 expression is independently associated with neo-osteogenesis in patients with chronic rhinosinusitis  Sakiko Oue, MBBS, Mahnaz Ramezanpour,

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Presentation on theme: "Increased IL-13 expression is independently associated with neo-osteogenesis in patients with chronic rhinosinusitis  Sakiko Oue, MBBS, Mahnaz Ramezanpour,"— Presentation transcript:

1 Increased IL-13 expression is independently associated with neo-osteogenesis in patients with chronic rhinosinusitis  Sakiko Oue, MBBS, Mahnaz Ramezanpour, BSc, PhD, Sathish Paramasivan, MBBS, BMedSci(Hons), Dijana Miljkovic, BMedPharmBiotech, PhD, Clare M. Cooksley, BSc(Hons), PhD, Ahmed Bassiouni, MBBS, PhD, Judy Ou, MBBS, Alkis J. Psaltis, MBBS(Hons), PhD, FRACS, Peter-John Wormald, MD, FRAC, FCS(SA), FRACS(Ed), MBChB, Sarah Vreugde, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 140, Issue 5, Pages e11 (November 2017) DOI: /j.jaci Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Differential mRNA expression (fold change) in patients with CRS without neo-osteogenesis (CRSwNeO−) and CRS with neo-osteogenesis patients (CRSwNeO+) normalized to 2 housekeeping genes (GAPDH and HPRT1) and relative to control subjects without CRS. Upregulation of ALPL, COL5A1, TNFA, CSF3, IL13, CCL13, and ITGAM in CRSwNeO+ patients, CRSwNeO− patients, or both relative to control subjects. *P < .05 and **P < .01. ns, Nonsignificant. Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 A-D, IL-13 immunohistochemistry of a representative CRSwNeO− patient (Fig 2, A) and CRSwNeO+ patient (Fig 2, B) with corresponding negative control staining (Fig 2, C and D). IL-13 is expressed in immune cells and in osteoblasts lining the bone (arrows in Fig 2, B, inlet). E and F, Alizarin Red assay demonstrates IL-13 dose-dependent osteoblastic mineralization (dose in nanograms per milliliter; Fig 2, E) and dose-response curve on a logarithmic scale (Pearson product-moment correlation = 0.586, P = .017; Fig 2, F; n = 2 duplicates). DA, Deactivated IL-13 (1000 ng/mL); NC, negative control. Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig E1 Scatterplot demonstrating the relationship between GOSS and IL13 expression and overlying correlation curve. The Pearson product-moment correlation value of 0.56 and P value of suggest a significant positive correlation between GOSS and IL-13 expression (n = 47). Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig E2 Representative images from the Alizarin Red mineralization assay. Red staining indicates calcium deposition by osteoblasts. A-D, When compared with control values (Fig E2, A), increasing doses of IL-13 (Fig E2, B, 10 ng/mL; Fig E2, C, 100 ng/mL; Fig E2, D, 1000 ng/mL) stimulated higher deposition by osteoblasts. E, Denatured IL-13 did not have any additional effect over the control. Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig E3 Micrograph of a representative CRSwNeO− patient, showing multiple bone fragments and selection of multiple hpfs measuring 0.035 mm2 beneath the epithelial layer. The inset shows an eosinophil (dashed red arrow points to an eosinophil with a 2-lobed nucleus and intense red eosin-positive staining) and 2 neutrophils (full arrows point to neutrophils harboring a multiple [2-5]–lobed nucleus and neutral pink staining). Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig E4 Comparison between eosinophil (A) and neutrophil (B) cell counts and basement membrane thickness (C) on histology in samples without neo-osteogenesis in CRSwNeO+ patients (CRSwNeO+ [non-neo-osteogenic area]) and in samples with neo-osteogenesis in the same CRSwNeO+ patients (CRSwNeO+ [neo-osteogenic area]) compared with patients who do not have evidence of neo-osteogenesis (CRSwNeO−) and control subjects without CRS. *P < .05. ns, Nonsignificant. Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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