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Differential cytokine induction by the human skin–associated autoallergen thioredoxin in sensitized patients with atopic dermatitis and healthy control.

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Presentation on theme: "Differential cytokine induction by the human skin–associated autoallergen thioredoxin in sensitized patients with atopic dermatitis and healthy control."— Presentation transcript:

1 Differential cytokine induction by the human skin–associated autoallergen thioredoxin in sensitized patients with atopic dermatitis and healthy control subjects  Susanne Hradetzky, PhD, Lennart Matthias Roesner, PhD, Annice Heratizadeh, MD, Reto Crameri, PhD, Mattia Garbani, MSc, Annika Scheynius, MD, PhD, Thomas Werfel, MD  Journal of Allergy and Clinical Immunology  Volume 135, Issue 5, Pages e5 (May 2015) DOI: /j.jaci Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Variable cytokine induction by hTrx and Mala s 13 in PBMCs from patients with AD who were sensitized or not to Malassezia species and healthy donors. A and B, PBMCs from patients with AD who were sensitized to Malassezia species (AD), healthy donors (ctrl), and patients with AD who were not sensitized to Malassezia species (AD ns) were stimulated with 2.5 μg/mL hTrx or Mala s 13 with or without IL-2 for 72 hours. Supernatants were analyzed for levels of IL-13 and IFN-γ (Fig 1, A) or IL-10 (Fig 1, B) by means of ELISA. C, PBMCs, isolated monocytes, or isolated CD4+ T cells from 5 healthy donors were stimulated with hTrx and Mala s 13 with or without IL-2 for 3 days. Supernatants were analyzed for levels of IL-10 by means of ELISA. D and E, PBMCs from 10 patients with AD who were sensitized to Mala s 13 were treated with PBS or stripping buffer for 10 minutes. Cells were analyzed for surface IgE through flow cytometry (Fig 1, D) or stimulated afterward as indicated for 3 days, and supernatants were analyzed for levels of IL-10 and IL-13 by using ELISA (Fig 1, E). Medians and minimums/maximums are shown. *P < .05 and **P < .01. ns, Not significant; S, stripping pretreatment. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig E1 hTrx is more efficient in IFN-γ and IL-10 induction in PBMCs from patients with AD than the autoallergen α-NAC. PBMCs from patients with AD who were sensitized or not sensitized to the respective allergens were stimulated with 2.5 μg/mL hTrx or 2.5 μg/mL α-NAC with or without IL-2 for 72 hours. Supernatants were analyzed for cytokine levels by means of ELISA. Medians are shown. *P < .05. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig E2 IgE sensitization to Mala s 13 or hTrx similarly affects IL-10 secretion in PBMCs from the respective sensitized patients with AD compared with control subjects. A, PBMCs from patients with AD who were sensitized to Mala s 13 (AD), healthy donors (ctrl), and patients with AD who were not sensitized to the respective antigen (AD ns) were stimulated with 2.5 μg/mL hTrx or 2.5 μg/mL Mala s 13 with or without IL-2 for 72 hours. Supernatants were analyzed for levels of IL-10 by means of ELISA. B, PBMCs from patients with AD who were sensitized to hTrx (AD) were compared with PBMCs from healthy donors (ctrl) and patients with AD who were not sensitized to the respective antigen (AD ns) under similar conditions, as described for Fig E2, A. Medians and minimums/maximums are shown. *P < .05 and **P < .01. ns, Not significant. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig E3 IL-10 is upregulated in PBMCs after monoclonal FcεR cross-linking. PBMCs from 4 patients with AD were stimulated with human myeloma IgE for 1 hour, washed, stimulated with rabbit anti-human IgE antibody for 1 hour, washed, and incubated for 24 hours. Supernatants were taken and analyzed for levels of IL-10 by means of ELISA. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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