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Thymic stromal lymphopoietin–activated invariant natural killer T cells trigger an innate allergic immune response in atopic dermatitis  Wen Hao Wu, PhD,

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Presentation on theme: "Thymic stromal lymphopoietin–activated invariant natural killer T cells trigger an innate allergic immune response in atopic dermatitis  Wen Hao Wu, PhD,"— Presentation transcript:

1 Thymic stromal lymphopoietin–activated invariant natural killer T cells trigger an innate allergic immune response in atopic dermatitis  Wen Hao Wu, PhD, Chang Ook Park, MD, Sang Ho Oh, MD, Hee Jung Kim, MD, Yeon Sook Kwon, MD, Byung Gi Bae, MD, Ji Yeon Noh, PhD, Kwang Hoon Lee, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 126, Issue 2, Pages e4 (August 2010) DOI: /j.jaci Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 iNKT cells in PBMCs of HCs and patients with AD. A, iNKT cells were identified as Vα24+/CD3+, Vα24+/Vβ11+, or 6B11+/Vβ11+ cells. B, The percentage of 6B11+/Vβ11+ iNKT cells in PBMCs of HCs (n = 10) and patients with mild (n = 5), moderate (n = 10), and severe AD (n = 10) was evaluated by using flow cytometry. FL, Fluorescence; ns, not significant. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 iNKT cell numbers increase in the lesional skin of patients with AD. A, Vα24+ iNKT cells were present among CD3+ T-cell infiltration in AD skin. Original magnification ×200. B, Representative immune-stained skin sections from HCs and patients with mild, moderate, and severe AD are shown. Original magnification ×100. Arrows indicate Vα24+ iNKT cells. C, The number of Vα24+ iNKT cells (cells/5 HPFs) in skin sections from HCs (n = 5) and mild (n = 5), moderate (n = 10), and severe AD skin lesions (n = 10). Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 TSLP receptor expression in iNKT cells. A, TSLPR and IL-7Rα protein expression in Vα24+ iNKT cells in normal PBMCs. Sorted 6B11+ iNKT cells from a normal donor expressed both TSLPR and IL-7Rα mRNA (B) and protein (C). FL, Fluorescence. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 A, Flow-cytometric analysis of IL-4 and IL-13 in Vα24+ iNKT cells in normal PBMCs treated with TSLP. B, Dose-dependent response to TSLP and neutralization effect of the anti-TSLPR Ab on Vα24+ iNKT cells. C, Confocal laser microscopy of purified iNKT cells from PBMCs treated with TSLP (a and d) and iNKT cells in lesional skin of patients with AD (g and j) expressing IL-4 (b and h) or IL-13 (e and k). Arrows indicate co-localization of iNKT cells with IL-4 (c and i) or IL-13 (f and l). Skin sections were counterstained with 4′,6-diamidino-2-phenylindole. Data shown are representative of 3 independent experiments. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 Cytokine expression in the iNKT cells (A), secreted cytokine levels in the supernatants from the iNKT cells (B), and cytokine expression in the iNKT cells cocultured with DCs (C) treated with TSLP or α-GalCer. Differing concentrations of TSLP were used (50 ng/mL for IL-4 or IFN-γ expression and 1 ng/mL for IL-13 expression). D, DCs expressed CD1d. Data shown are representative of 3 independent experiments. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 5 Cytokine expression in the iNKT cells (A), secreted cytokine levels in the supernatants from the iNKT cells (B), and cytokine expression in the iNKT cells cocultured with DCs (C) treated with TSLP or α-GalCer. Differing concentrations of TSLP were used (50 ng/mL for IL-4 or IFN-γ expression and 1 ng/mL for IL-13 expression). D, DCs expressed CD1d. Data shown are representative of 3 independent experiments. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Serum IgE levels did not correlate with TSLP expression, iNKT cell levels, or the EASI scores of patients with AD. A, Correlation between serum IgE levels and the percentage of 6B11+/Vβ11+ iNKT cells in PBMCs of patients with AD (n = 25). Correlation between serum IgE levels and the number of Vα24+ iNKT cells (B) or the staining intensity of TSLP (C) in skin lesions of patients with AD (n = 25). D, Correlation between serum IgE levels and the EASI scores of patients with AD (n = 25). Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 TSLP expression in lesional skin and sera of patients with AD
TSLP expression in lesional skin and sera of patients with AD. A, Representative immune-stained skin sections of HCs and patients with AD are shown. Original magnification ×100. B, The intensity of TSLP staining was graded visually on a scale from 0 (no staining) to 6 (the most intense staining) for skin sections from HCs (n = 5) and mild (n = 5), moderate (n = 10), and severe AD skin lesions (n = 10). C, TSLP protein in the sera of HCs and patients with AD was evaluated by using ELISA. ns, Not significant. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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