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Targeting Fel d 1 to FcγRI induces a novel variation of the TH2 response in subjects with cat allergy  Kathryn E. Hulse, BS, Amanda J. Reefer, MS, Victor.

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Presentation on theme: "Targeting Fel d 1 to FcγRI induces a novel variation of the TH2 response in subjects with cat allergy  Kathryn E. Hulse, BS, Amanda J. Reefer, MS, Victor."— Presentation transcript:

1 Targeting Fel d 1 to FcγRI induces a novel variation of the TH2 response in subjects with cat allergy  Kathryn E. Hulse, BS, Amanda J. Reefer, MS, Victor H. Engelhard, PhD, Shama M. Satinover, MS, James T. Patrie, MS, Martin D. Chapman, PhD, Judith A. Woodfolk, MBChB, PhD  Journal of Allergy and Clinical Immunology  Volume 121, Issue 3, Pages e4 (March 2008) DOI: /j.jaci Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 PBMC proliferation to H22–Fel d 1. PBMCs were stimulated with allergen and proliferation measured on day 7 (n = 7 per group). Values correspond to mean counts per minute + SDs. ∗P < .01; ∗∗P < Subjects with cat allergy: P < .04 for unstim versus all other conditions. Modified TH2 responders: P < .02 for unstim versus Fel d 1 (10 μg/mL) and H22–Fel d 1 (1 and 10 μg/mL). Unstim, Unstimulated. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Effects of H22–Fel d 1 on MDDCs. MDDCs from subjects with cat allergy (n = 6) were pulsed with antigen (24 hours) and analyzed for expression of surface antigens and cytokine secretion. A, Representative data showing expression of CD64. B, Effects of different stimuli on maturation markers and CD64. ∗P < .01. C, Effects of different stimuli on cytokine secretion. Values represent the means + SDs. ∗P < .05; ∗∗P < .01. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Changes in cytokine-positive CD4+ T cells induced by H22–Fel d 1 compared with Fel d 1. CD4+ T cells were cocultured with antigen-pulsed MDDCs for 10 days, and the percentage of total CD4+ T cells that were IFN-γ+, IL-5+, and IL-10+ was analyzed by flow cytometry. Experiments were performed in subjects with cat allergy (n = 10), modified TH2 (n = 5), and control (n = 5) subjects. Boxes represent medians and interquartile ranges. ∗Significant after adjusting for multiple comparisons. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Representative data showing changes in cytokine-positive CD4+ T cells induced by H22–Fel d 1. Flow cytometry dot plots are shown for 1 of 10 patients with allergy. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 Summary of cytokine-positive CD4+ T-cell subtypes induced by H22–Fel d 1. A, Comparison of 6 CD4+ T-cell subtypes induced by Fel d 1 and H22–Fel d 1 in subjects with cat allergy (n = 10), modified TH2 (n = 5), and control (n = 5) subjects. B, Comparison of CD4+ T-cell subtypes induced by H22–Fel d 1 with Der p 1 in subjects cosensitized to cat and dust mite allergens (n = 5). Bars represent the geometric mean of the percentage of total CD4+ T cells for each subtype. ∗Significant after adjusting for multiple comparisons. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 Effect of blocking IL-10 on cytokine-positive CD4+ T cells induced by H22–Fel d 1. CD4+ T cells were cocultured with MDDCs pulsed with H22–Fel d 1 in the presence of a blocking anti–IL-10 receptor mAb or isotype control (n = 5 subjects with cat allergy). A, Representative flow cytometry dot plots. B, Frequency of IL-5+ and IFN-γ+ CD4+ T cells in the presence or absence of anti–IL-10 receptor (IL-10R) mAb for each subject. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Changes in cytokine-positive CD4+ T-cell subtypes induced by H22–Fel d 1 compared with Fel d 1. CD4+ T cells were cocultured with MDDCs pulsed with Fel d 1 or H22–Fel d 1 for 10 days, and the percentage of the following 6 CD4+ T-cell subtypes was analyzed by flow cytometry: (1) IFN-γ+ only; (2) IL-5+ only; (3) IFN-γ+IL-5+; (4) IL-10+ only; (5) IL-10+IFN-γ; and (6) IL-10+IL-5+. Experiments were performed in subjects with cat allergy (n = 10), modified TH2 (n = 5), and control (n = 5) subjects. Boxes represent medians and interquartile ranges. ∗Significant after adjusting for multiple comparisons. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Comparison of T-cell repertoires induced by H22–Fel d 1 and Der p 1
Comparison of T-cell repertoires induced by H22–Fel d 1 and Der p 1. CD4+ T cells were cocultured with MDDCs pulsed with H22–Fel d 1 or Der p 1 for 10 days, and the percentage of the following 6 CD4+ T-cell subtypes was analyzed by flow cytometry: (1) IFN-γ+ only; (2) IL-5+ only; (3) IFN-γ+IL-5+; (4) IL-10+ only; (5) IL-10+IFN-γ; and (6) IL-10+IL-5+. Experiments were performed in 5 subjects who were cosensitized to cat and dust mite allergens. Boxes represent medians and interquartile ranges. ∗Significant after adjusting for multiple comparisons. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 Effect of blocking IL-10 on cytokine-positive CD4+ T cells induced by H22–Fel d 1. CD4+ T cells were cocultured with MDDCs pulsed with H22–Fel d 1 in the presence of a blocking anti–IL-10 receptor mAb or isotype control. Flow cytometry dot plots showing effects of IL-10 blockade on IL-10+ and IFN-γ+ cells (representative of 5 subjects with cat allergy). Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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