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IL-17 in atopic eczema: Linking allergen-specific adaptive and microbial-triggered innate immune response  Kilian Eyerich, MD, Davide Pennino, PhD, Claudia.

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Presentation on theme: "IL-17 in atopic eczema: Linking allergen-specific adaptive and microbial-triggered innate immune response  Kilian Eyerich, MD, Davide Pennino, PhD, Claudia."— Presentation transcript:

1 IL-17 in atopic eczema: Linking allergen-specific adaptive and microbial-triggered innate immune response  Kilian Eyerich, MD, Davide Pennino, PhD, Claudia Scarponi, PhD, Stefanie Foerster, PhD,, Francesca Nasorri, PhD, Heidrun Behrendt, MD, Johannes Ring, MD, PhD, Claudia Traidl-Hoffmann, MD, Cristina Albanesi, PhD, Andrea Cavani, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 123, Issue 1, Pages e4 (January 2009) DOI: /j.jaci Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Distinct populations of IL-17–producing T cells in the skin infiltrate of a positive APT reaction. A-C, Intracellular cytokine staining of skin-derived lymphocytes with IL-4/IFN-γ (Fig 1, A), IL-4/IL-17 (Fig 1, B), and IL-17/IL-22 (Fig 1, C). D, Gating on IL-17+ cells after triple staining with IL-4, IFN-γ, and IL-17. Percentage of cells is indicated for each quadrant. Shown is 1 representative experiment. APC, Allophycocyanin. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 IL-17+ Der p 1–specific T-cell clones secrete IL-17 on stimulation with SEB. A and B, Dose-dependent secretion of IL-17/IL-4/IFN-γ (left panel) and addition of cytokines and SEB (right panel). Type of APC: immature DCs. Error bars indicate SDs of 1 representative experiment performed in triplicate. C and D, Relative cytokine profile (mean percentage of 3 independent experiments) of clones shown in A and B. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 SEB activates T cells through a TCR-dependent mechanism. Clone 3 was stimulated with increasing amounts of SEB in the presence of 5000 immature DCs for 48 hours. Experimental end points were proliferation (A) and IL-17 secretion (B). Addition of anti-HLA-DP/DQ/DR almost completely abrogated SEB stimulation (hatched bars). Error bars indicate SEMs of 2 independent experiments. iDC, Immature DCs. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 T cell–derived IL-17 induces HBD-2 in primary human keratinocytes, and IL-4 and IL-13 partially block this effect. Induction of HBD-2 mRNA (A) and protein (B) in AE and healthy keratinocytes in response to recombinant cytokines and to cell-free supernatant obtained from stimulated T-cell clones (C) is shown. Error bars indicate SEMs of 3 independent experiments. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 SEB induces expression of IL-17 and HBD-2 in vivo. Levels of IL-17 (A), IFN-γ (B), IL-4 (C), and IL-10 (D) in suction blister fluids as detected with Luminex analysis. E and F, Relative induction of HBD-2 mRNA (E) and protein content (F). Error bars indicate SDs of 1 experiment performed in triplicate. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Cytokine pattern of IL-17–secreting T-cell clones confirms the existence of TH17, TH1/IL-17, and TH2/IL-17 subpopulations in APT reactions. Cytokine secretion after 48-hour PMA/ionomycin stimulation of all 34 identified IL-17–producing T-cell clones is shown. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Stimulation of Der p 1–specific T-cell clones with mature DCs results in high IL-17 secretion on SEB but not on allergen stimulation. A and B, Dose-dependent secretion of IL-17 and IL-4 (left panel) and addition of cytokines and SEB (right panel) are shown. Type of APC: mature DCs (mDC). Error bars indicate SDs of 1 representative experiment performed in triplicate. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Stimulation ofDer p 1–specific T-cell clones with CD14+ monocytes results in high IL-17 secretion on SEB but not on allergen stimulation. A and B, Dose-dependent secretion of IL-17 and IL-4 (left panel) and addition of cytokines and SEB (right panel) are shown. Type of APC: CD14+ monocytes (monos). Error bars indicate SDs of 1 representative experiment performed in triplicate. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 Varying concentrationsof Der p 1 have little effect on IL-17 secretion by TH2/IL-17 clones: A, 1 μg/mL Der p 1; B, 5 μg/mL Der p 1. iDC, Immature DCs. Error bars indicate SEMs from 3 independent experiments with clone 3. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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