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Decreases in human dendritic cell–dependent TH2-like responses after acute in vivo IgE neutralization  John T. Schroeder, PhD, Anja P. Bieneman, BS, Kristin.

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Presentation on theme: "Decreases in human dendritic cell–dependent TH2-like responses after acute in vivo IgE neutralization  John T. Schroeder, PhD, Anja P. Bieneman, BS, Kristin."— Presentation transcript:

1 Decreases in human dendritic cell–dependent TH2-like responses after acute in vivo IgE neutralization  John T. Schroeder, PhD, Anja P. Bieneman, BS, Kristin L. Chichester, MS, Robert G. Hamilton, PhD, HuiQing Xiao, MD, Sarbjit S. Saini, MD, Mark C. Liu, MD  Journal of Allergy and Clinical Immunology  Volume 125, Issue 4, Pages e6 (April 2010) DOI: /j.jaci Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 FcεRIα and surface IgE on human blood DC subtypes are downregulated after omalizumab administration. BDCA2+ pDCs and BDCA1+CD19- mDCs within PBMC suspensions were analyzed pre (baseline) and post (∼d105) treatment with active (n = 12) drug or placebo (n = 4). Shown are the median, IQR, and lower/upper extreme fractional values posttreatment relative to baseline (see Figs E2 and E3 for specific nMFI values). ∗P = .0005; #P < Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 FcεRIα correlates with free IgE levels postomalizumab treatment. Free IgE serum levels were determined posttreatment (∼d105). Shown are percentages of the total IgE at the same time point (A and C) and as the total IgE at baseline (B and D) vs fractional (post/pre) values for FcεRIα on both pDCs (A and B) and mDCs (C and D). Open circles denote placebo subjects. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 DC-dependent T-cell proliferation to allergen is inhibited with omalizumab administration. DC subtypes were cultured with CD4+ T cells as described in Methods. Shown are the median, IQR, and lower/upper extreme cpms for pDC/CD4 (A and B) and mDC/CD4 (C and D) cocultures using cells isolated from active (n = 11) and placebo (n = 4) subjects pre and posttreatment. ∗P = .001. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Cytokine secretion in pDC/T-cell cocultures after omalizumab administration. BDCA4+ DC and CD4+ T-cells were isolated pre and posttreatment and cocultured with 100 and 500 BAU/mL cat extract for 96 hours before assaying supernatants for common TH2-like (A) and TH1-like (B) cytokines by using multiplexing. Shown are the median, IQR, and lower/upper extreme pg/mL values for the indicated cytokines detected among the active (n = 11) and placebo (n = 4) groups. ∗P ≤ .05. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Gating parameters used in detecting mDC (BDCA1+CD19- cells) for determination of FcεRIα and surface IgE expression. Representative data. A, Forward and side scatter analysis of a BDC suspension (ie, PBMCs). B, Staining parameters to identify BDCA1+CD19- cells (ie, mDC). C, Staining for FcεRIα on mDCs. Shaded area represents staining with isotype control antibody. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 FcεRIα expression on pDCs and mDCs correlates with serum total IgE and surface IgE expression. BDCA2+ pDCs and BDCA1+CD19- mDCs within PBMC suspensions were analyzed at baseline for FcεRIα expression and surface IgE staining by using flow cytometry. Total IgE levels were determined by the ImmunoCap-250 assay. Shown are regression analyses with r2 and P values. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 FcεRIα and surface IgE expression on pDCs are downregulated after omalizumab administration. PBMCs prepared pre (d0) and post (∼d105) treatment from subjects receiving omalizumab (Act) or placebo (PL) were analyzed by flow cytometry for BDCA2+ cells (pDC) coexpressing FcεRIα (A and B) and surface IgE (C and D). Shown are results for individual subjects enrolled. Solid black bars denote mean values for the group indicated. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 FcεRIα and surface IgE expression on mDCs are downregulated after omalizumab administration. PBMCs prepared pre (d0) and post (∼d105) treatment from subjects receiving omalizumab (Act) or placebo (PL) were analyzed by flow cytometry for BDCA1+CD19- cells (mDC) coexpressing FcεRIα (A and B) and surface IgE (C and D). Shown are results for individual subjects enrolled. Solid black bars denote mean values for the group indicated. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 FcεRIα expression on pDC after omalizumab administration versus the secretion of IL-5 (A) and IL-13 (B) in pDC/CD4+ T-cell cocultures. Fractional responses posttreatment relative to baseline values are plotted for subjects receiving omalizumab (n = 11). Cytokine values are from those induced with the 100 BAU/mL concentration of cat extract. Open circles with arrows represent presumed outlier results, which were not included in regression analyses. Journal of Allergy and Clinical Immunology  , e6DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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