Cross-talk between human mast cells and epithelial cells by IgE-mediated periostin production in eosinophilic nasal polyps Dae Woo Kim, MD, PhD, Marianna.

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Cross-talk between human mast cells and epithelial cells by IgE-mediated periostin production in eosinophilic nasal polyps Dae Woo Kim, MD, PhD, Marianna Kulka, PhD, Ara Jo, PhD, Kyoung Mi Eun, BSc, Narcy Arizmendi, PhD, Brian P. Tancowny, PhD, Seung-No Hong, MD, Jung Pyo Lee, MD, PhD, Hong Ryul Jin, MD, PhD, Richard F. Lockey, MD, Dong-Kyu Kim, MD, PhD, Seong H. Cho, MD Journal of Allergy and Clinical Immunology Volume 139, Issue 5, Pages 1692-1695.e6 (May 2017) DOI: 10.1016/j.jaci.2016.09.026 Copyright © 2016 Terms and Conditions

Fig 1 Cross-talk between human mast cells and epithelial cells in eosinophilic nasal polyps. A and B. IHC detection of periostin was performed within the NP tissues from patients with CRSwNP (eosinophil, asterisk; mast cell, red arrowhead; fibroblast, black arrow). Black scale bar = 10 μm. C, Representative double immunostaining (red, tryptase; blue, periostin). Arrows indicate double-positive immune cells (×1000). Black scale bar = 10 μm. D, Counts of double-positive cells in NE-NPs and E-NPs (n = 5 for each group). E, Periostin mRNA expression was measured by quantitative RT-PCR in LAD2 cells stimulated with IgE, IL-4, IL-13, and TNF-α (n = 9 for each group). F, Periostin protein expression stimulated by IgE (n = 3 for each group). G, Number of integrin αV–positive epithelial cells per 100 cells was determined and averaged from 3 different areas of epithelium (n = 10 for control-UP, n = 10 for CRSsNP-UP, n = 10 for CRSwNP-UP, and n = 37 for CRSwNP-NP) in IHC assays of tissue sections. The number of integrin αV–positive inflammatory cells was counted from 5 of the densest areas (hpf; ×400) and averaged for each group. H, Correlation between integrin αV and TSLP at the protein level. I, Correlation between periostin and TSLP at the protein level. J, Periostin-induced TSLP production in NHBE cells after TH2 cytokine and poly(I:C) treatment (n = 3 for each group). K and L, Periostin-induced TSLP production was reversed by antiintegrin αVβ3 (Fig 1, K) and antiintegrin αVβ5 (Fig 1, L) (n = 3 for each group). IHC, Immunohistochemical staining; NHBE, normal human bronchial epithelial; SA, streptavidin. *P < .05, **P < .01, ***P < .001, and ****P < .0001 using the Mann-WhitneyU test. Journal of Allergy and Clinical Immunology 2017 139, 1692-1695.e6DOI: (10.1016/j.jaci.2016.09.026) Copyright © 2016 Terms and Conditions

Fig E1 Expression of periostin in E-NPs. A, Relative periostin mRNA expression was measured by quantitative RT-PCR in tissues from each group (n = 22 for control-UP, n = 62 for CRSsNP-UP, n = 68 for CRSwNP-UP, and n = 67 for CRSwNP-NP). B, Protein levels of periostin were measured by ELISA (n = 7 for control-UP, n = 11 for CRSsNP-UP, n = 12 for CRSwNP-UP, and n = 43 for CRSwNP-NP). C, Correlation between Lund-MacKay computed tomography score and protein levels of periostin in different subtypes of CRSwNP. Journal of Allergy and Clinical Immunology 2017 139, 1692-1695.e6DOI: (10.1016/j.jaci.2016.09.026) Copyright © 2016 Terms and Conditions

Fig E2 Correlation between periostin expression and major cytokines in NPs. A, IL-5 (n = 18 for E-NPs and n = 16 for NE-NPs). B, CCL-11 (n = 18 for E-NPs and n = 16 for NE-NPs).C, IL-17A (n = 18 for E-NPs and n = 16 for NE-NPs). D, IFN-γ (n = 18 for E-NPs and n = 26 for NE-NPs). E, Periostin levels in comparison with total IgE in NP homogenates (n = 16 for E-NPs and n = 15 for NE-NPs).CCL-11, C-C motif ligand 11. Journal of Allergy and Clinical Immunology 2017 139, 1692-1695.e6DOI: (10.1016/j.jaci.2016.09.026) Copyright © 2016 Terms and Conditions

Fig E3 Schematic depiction of periostin-mediated cross-talk between MCs and epithelium. Journal of Allergy and Clinical Immunology 2017 139, 1692-1695.e6DOI: (10.1016/j.jaci.2016.09.026) Copyright © 2016 Terms and Conditions