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IgG4 production is confined to human IL-10–producing regulatory B cells that suppress antigen-specific immune responses  Willem van de Veen, MSc, Barbara.

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Presentation on theme: "IgG4 production is confined to human IL-10–producing regulatory B cells that suppress antigen-specific immune responses  Willem van de Veen, MSc, Barbara."— Presentation transcript:

1 IgG4 production is confined to human IL-10–producing regulatory B cells that suppress antigen-specific immune responses  Willem van de Veen, MSc, Barbara Stanic, PhD, Görkem Yaman, MD, Marcin Wawrzyniak, MSc, Stefan Söllner, Sci Tec, Deniz G. Akdis, Beate Rückert, Sci Tec, Cezmi A. Akdis, MD, Mübeccel Akdis, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 131, Issue 4, Pages (April 2013) DOI: /j.jaci Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Characterization of IL-10− and IL-10+ B cells. A, Sorting of IL-10− and IL-10+ B cells. IL-10 expression was confirmed by using real-time PCR. B, Gene array result showing upregulated or downregulated surface marker genes in IL-10− versus IL-10+ B cells. C, mRNA expression (relative to IL-10−) of selected surface markers (n = 6). D, CD25, CD71, CD274, and CD73 surface staining (representative example of 3). *P < .05 and **P < .01. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Sorting of cell populations from resting B cells based on expression of CD73, CD25, and CD71. A, Staining of CD25 and CD71 on CD73− and CD73+ B cells (n = 3; 1 representative example is shown). B, Sorted B-cell populations were reanalyzed to assess purity. C, Sorted B-cell populations (n = 3, 5 × 104 cells in 100 μL) were stimulated for 48 hours, and IL-10 levels were measured in supernatants. *P < .05. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 BR1 cells suppress antigen-specific proliferation. A, Cocultures of carboxyfluorescein succinimidyl ester (CFSE)–labeled PBMCs with IL-10+ or IL-10− B cells (ratio, 1 B cell to 25 PBMCs) with blocking anti–IL-10R or isotype control (i.c.) mAbs. rIL-10 was used as a control. Histograms show a representative example. Numbers indicate the percentage dividing CD4+ cells. B, Cocultures of PBMCs with CD73, CD25, and CD71+/− B cells. Combined data from 4 independent experiments are shown. *P < .05 and **P < .01. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Immunoglobulin production by BR1 cells. A, IgG1, IgG4, IgA, and IgE mRNA expression (relative to unstimulated cells) in total B cells. Lower panel, IgG1, IgG4, and IgA levels measured in supernatants (n = 5, 3 independent experiments). B, IgG1, IgG4, IgA, and IgE mRNA expression (average IL-10− expression = 1) in IL-10− and IL-10+ B cells. Connected dots represent individual donors. C, Secreted IgG1, IgG4, and IgA measured from IL-10− and IL-10+ cells derived from CD27− and CD27+ B cells (n = 4). Fold increase is relative to the average of IL-10−. IgE protein was not detectable. *P < .05 and **P < .01. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 Allergen-specific B cells from tolerant subjects upregulate IL-10 and IgG4. A, Enrichment of PLA-specific B cells from beekeeper-derived peripheral CD19+ cells. B, Direct ex vivo analysis of immunoglobulin and IL-10 mRNA expression (relative to PLA− cells, n = 6). Horizontal bars indicate medians. C, IL-10 and PLA staining in B cells from beekeepers (n = 7) or patients with bee venom allergy (n = 4) after 3 days of TLR9-L stimulation. Numbers in dot plots indicate means ± SDs. D, PLA-specific serum IgE/IgG4 ratio. *P < .05, **P < .01, and ***P < .001. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Journal of Allergy and Clinical Immunology 2013 131, 1204-1212DOI: (10
Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Journal of Allergy and Clinical Immunology 2013 131, 1204-1212DOI: (10
Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Fig E1 Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 Fig E2 Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

11 Fig E3 Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

12 Fig E4 Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

13 Fig E5 Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2013 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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