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CD23 surface density on B cells is associated with IgE levels and determines IgE- facilitated allergen uptake, as well as activation of allergen-specific.

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Presentation on theme: "CD23 surface density on B cells is associated with IgE levels and determines IgE- facilitated allergen uptake, as well as activation of allergen-specific."— Presentation transcript:

1 CD23 surface density on B cells is associated with IgE levels and determines IgE- facilitated allergen uptake, as well as activation of allergen-specific T cells  Regina Selb, PhD, Julia Eckl-Dorna, MD, PhD, Alina Neunkirchner, PhD, Klaus Schmetterer, MD, PhD, Katharina Marth, MD, Jutta Gamper, BSc, Beatrice Jahn-Schmid, PhD, Winfried F. Pickl, MD, Rudolf Valenta, MD, Verena Niederberger, MD  Journal of Allergy and Clinical Immunology  Volume 139, Issue 1, Pages e4 (January 2017) DOI: /j.jaci Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Associations of numbers of CD23 molecules per B cell and percentages of CD23+ B cells with skin sensitivity to grass pollen allergens and total and grass pollen–specific IgE levels. Numbers of CD23 molecules per CD23+ B cells (A-C) or percentages of CD23+ B cells (D-F) are displayed on the y-axis. Total IgE levels (Fig 1, A and D), timothy grass pollen–specific IgE levels (Fig 1, B and E), or sizes of grass pollen–induced wheal reactions (Fig 1, C and F) are plotted on the x-axis. *P values of less than .05. **P values of less than .01. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Dependence of allergen uptake on CD23 cell-surface density. A, Confocal microscopy of 3 EBV-transformed B-cell lines expressing different numbers of CD23 molecules on their surfaces (I, 3.5 × 105; II, 1 × 105; and III, 5 × 104 CD23 molecules per cell). Cells stained with anti-CD19 are shown in pink, whereas allergen (Bet v 1 trimer) was labeled in green with DyLight 488, preincubated with allergen-specific IgE, and then exposed to the cells. The pictures marked overlay show an overlay of anti-CD19 and allergen staining. B and C, Binding (Fig 2, B) and uptake (Fig 2, C) of a fluorescent allergen-IgE complex by CD23-expressing EBV-transformed B-cell lines (I, II, and III) shown by using flow cytometry. CD23 expression was measured before and after 3.5 hours of incubation with IgE-allergen complexes. Different symbols (circles, I; squares, II: and triangles, III) represent the 3 different B-cell lines. Four independent experiments performed on 4 different study days are displayed. Each data point represents the mean of triplicate experiments done on the same study day. Background fluorescence measured after incubation with allergen alone was subtracted. Dotted lines, Trend lines of correlations. D and E, Binding (Fig 2, D) and uptake (Fig 2, E) of allergen-IgE complexes was blocked by an anti-CD23 antibody but not by the matching isotype control (IgG1). Solid triangles represent blocking by anti-CD23, and open circles show cells incubated with the isotype control. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 2 Dependence of allergen uptake on CD23 cell-surface density. A, Confocal microscopy of 3 EBV-transformed B-cell lines expressing different numbers of CD23 molecules on their surfaces (I, 3.5 × 105; II, 1 × 105; and III, 5 × 104 CD23 molecules per cell). Cells stained with anti-CD19 are shown in pink, whereas allergen (Bet v 1 trimer) was labeled in green with DyLight 488, preincubated with allergen-specific IgE, and then exposed to the cells. The pictures marked overlay show an overlay of anti-CD19 and allergen staining. B and C, Binding (Fig 2, B) and uptake (Fig 2, C) of a fluorescent allergen-IgE complex by CD23-expressing EBV-transformed B-cell lines (I, II, and III) shown by using flow cytometry. CD23 expression was measured before and after 3.5 hours of incubation with IgE-allergen complexes. Different symbols (circles, I; squares, II: and triangles, III) represent the 3 different B-cell lines. Four independent experiments performed on 4 different study days are displayed. Each data point represents the mean of triplicate experiments done on the same study day. Background fluorescence measured after incubation with allergen alone was subtracted. Dotted lines, Trend lines of correlations. D and E, Binding (Fig 2, D) and uptake (Fig 2, E) of allergen-IgE complexes was blocked by an anti-CD23 antibody but not by the matching isotype control (IgG1). Solid triangles represent blocking by anti-CD23, and open circles show cells incubated with the isotype control. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 3 IgE-facilitated T-cell activation is dependent on CD23 surface density. EBV-immortalized B cells were sorted according to their CD23 expression. A, Correlation between numbers of CD23 molecules on B cells (x-axis) and T-cell activation (y-axis, luciferase activity in arbitrary units) induced by IgE–Bet v 1 immune complexes. Four independent experiments on 4 different study days are displayed. B, Activation of Bet v 1–specific T cells (y-axis) after incubation of sorted EBV-immortalized B cells (white, low expression; gray, intermediate expression; and black, high expression) with IgE–Bet v 1 immune complexes, Bet v 1–specific IgE and the unrelated allergens Art v 1 or Bet v 1 alone, respectively. Data from 2 independent experiments showed similar results and were summarized. C, IgE-facilitated T-cell activation was blocked by preincubation of B cells with an anti-CD23 antibody (anti-CD23) but not by an isotype control antibody (IgG1). Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig E1 CD23+ B cells mainly have a naive IgD+ phenotype. Data from 1 representative patient are shown. Of the 78.1% CD23+ B cells, 94.4% were IgD+ naive cells. The population of CD23− B cells consisted of 21.9% of the cells, of which 49.6% were IgD+ naive cells and 9.17% were CD27+IgD− memory B cells. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig E2 Changes of CD23 surface density on B cells after addition of IgE. Shown are percentage changes of CD23 density on B cells in PBMCs from allergic (black) and nonallergic (white) subjects cultured after addition of an IgE mAb (1 μg/mL IgE) for 6 days compared with untreated cells. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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