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Aarti Shikotra, BSc, David F. Choy, BSc, Chandra M

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1 Increased expression of immunoreactive thymic stromal lymphopoietin in patients with severe asthma 
Aarti Shikotra, BSc, David F. Choy, BSc, Chandra M. Ohri, MD, Emma Doran, BSc, Claire Butler, MBBCh, Beverley Hargadon, BSc, Maria Shelley, BA, Alexander R. Abbas, PhD, Cary D. Austin, MD, PhD, Janet Jackman, MSc, Lawren C. Wu, PhD, Liam G. Heaney, MD, Joseph R. Arron, MD, PhD, Peter Bradding, DM  Journal of Allergy and Clinical Immunology  Volume 129, Issue 1, Pages e9 (January 2012) DOI: /j.jaci Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 TSLP immunostaining is increased in asthmatic patients. A and B, TSLP immunostaining in the airway epithelium from a patient with asthma (Fig 1, A) and inhibition by recombinant TSLP (Fig 1, B). C and D, TSLP immunostaining in the lamina propria from an asthmatic subject (Fig 1, C) and inhibition by recombinant TSLP (Fig 1, D). E and F, Extent of TSLP immunostaining in airway epithelium assessed based on threshold measurements and scoring, respectively. ∗P < .05, ∗∗P = .0001, and ∗∗∗P = compared with healthy control subjects. G, Extent of TSLP immunostaining in the lamina propria. ∗P = compared with healthy control subjects. Immunohistology is shown at ×400 magnification. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 IL-13 expression in the airway epithelium and lamina propria. A, C, D, and F, IL-13 immunoreactivity in the airway epithelium and lamina propria from 4 patients with severe asthma. In Fig 2, D, arrows demonstrate IL-13+ pseudocolumnar epithelial cells, and arrowheads highlight IL-13+ nonepithelial cells. B, An immunoabsorption control with recombinant human IL-13 in a section adjacent to Fig 2, A. E and F, Sequential sections demonstrating colocalization of IL-13 to CD45+ cells (arrows; ×400 magnification). G, The density of IL-13+ nonepithelial cells within the airway epithelium. ∗P = H, The density of IL-13+ cells within the airway lamina propria. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Relationships between IL-13, IL-4, and TSLP expression in the lamina propria in asthmatic patients. IL-13 immunoreactivity in the lamina propria was scored as absent or present; these categorical descriptions of IL-13 immunoreactivity were compared against IL-4 (A) and TSLP (B) immunoreactivity in the lamina propria. Gray symbols, Mild asthma; white symbols, moderate asthma; black symbols, severe asthma. ∗P = .014 and ∗∗P = .022. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Bronchial biopsy specimen gene expression of TSLP, TH2 signature score, and eosinophil count are intercorrelated. A, A quantitative score for TH2 signature score was calculated from gene expression microarrays (see the Methods section in this article’s Online Repository). Relative expression is represented by means of heat map, where patient samples (columns) are organized left to right by increasing the TH2 signature score, as indicated by the adjacent bar plot (top). Genes (rows) are organized by principal component analysis loadings as indicated by adjacent dot plots (left). B-D, All pairwise comparisons of bronchial biopsy TSLP expression, TH2 signature scores, and eosinophil counts are represented by scatterplots (Spearman rank, all comparisons P < .05). Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig E1 A and B, Positive control immunostaining for TSLP in human tonsil tissue (Fig E1, A) and negative immunostaining in tonsil tissue with sheep IgG isotype control (Fig E1, B; 7.5 μg/mL). C, Negative immunostaining with sheep IgG isotype control in a bronchial biopsy specimen from a patient with severe asthma. D-F, Immunostaining in tonsil tissue for CD83, OX40, and OX40L, respectively. G, Immunostaining for CD1a in normal bronchial epithelium and lamina propria. H, Immunostaining for CD83 in normal bronchial lamina propria. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig E2 Colocalization of TSLP to mast cells and macrophages within the lamina propria. A, Two sequential 2-μm sections demonstrating colocalization of mast cell tryptase and TSLP (arrows). Lines demonstrate local landmarks for reference. B, The percentage of macrophages and mast cells expressing TSLP and vice versa in a subset of 6 patients with severe asthma. Immunohistology is shown at ×400 magnification. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig E3 A and B, The density of IL-4+ cells in the bronchial epithelium and lamina propria, respectively. C, The density of IL-13+ epithelial cells within the airway epithelium. Journal of Allergy and Clinical Immunology  , e9DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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