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Bruton tyrosine kinase mediates TLR9-dependent human dendritic cell activation  Vassilios Lougaris, MD, Manuela Baronio, PhD, Massimiliano Vitali, PhD,

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Presentation on theme: "Bruton tyrosine kinase mediates TLR9-dependent human dendritic cell activation  Vassilios Lougaris, MD, Manuela Baronio, PhD, Massimiliano Vitali, PhD,"— Presentation transcript:

1 Bruton tyrosine kinase mediates TLR9-dependent human dendritic cell activation 
Vassilios Lougaris, MD, Manuela Baronio, PhD, Massimiliano Vitali, PhD, Giacomo Tampella, PhD, Marco Cattalini, MD, Laura Tassone, PhD, Annarosa Soresina, MD, Raffaele Badolato, MD, PhD, Alessandro Plebani, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 133, Issue 6, Pages e4 (June 2014) DOI: /j.jaci Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Human DCs activation upon LPS or CpG stimulation in the presence or absence of BTK. A, Summarized data for upregulation of CD86, CD83, CD80, and HLA-DR upon LPS or CpG stimulation in human DCs from healthy donors (Hd) (n = 13) and patients with XLA (Pt) (n = 13). B, Histograms that depict the mean fluorescence intensity of CD86, CD83, CD80, and HLA-DR upon LPS (black) and CpG (red) stimulation in DCs from a healthy control (BTK+/+) (upper panel) and an XLA patient (BTK−/−) (lower panel). Cells cultured in medium alone were used as control (gray) (representative experiment). Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Human DCs cytokine induction upon LPS or CpG stimulation in the presence or absence of BTK. Cytokine mRNA levels for IL6, IL-12, TNF-α, and IL-10 upon LPS or CpG stimulation from human DCs from healthy donors (Hd) (n = 10) and patients with XLA (Pt) (n = 10). Bar graphs show mean values of 2 (+ SD) technical replicates. P values were calculated by using the unpaired Student t test (*P = ; **P = ). ns, Not significant. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 STAT1/3 upregulation in human DCs upon LPS or CpG stimulation in the presence or absence of BTK. STAT1 and STAT3 mRNA levels were measured in DCs from healthy donors (n = 10) and patients with XLA (n = 10) (BTK−/−). Bar graphs show values of 2 (+ SD) technical replicates. P values were calculated by using unpaired Student t test (**P = ). ns, Not significant. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Human DC activation upon LPS or CpG stimulation in the presence or blockade of BTK. A, Summarized data for upregulation of CD86, CD83, CD80, and HLA-DR upon LPS or CpG stimulation in human DCs from 11 healthy donors (Hd) in the presence or absence of the specific BTK inhibitor ibrutinib. B, Representative histograms, depicting the mean fluorescence intensity of CD86, CD83, CD80, and HLA-DR expression upon LPS (black) and CpG (red) stimulation in DCs from healthy donors in the presence (Hd + irutinib) or absence (Hd) of specific BTK inhibitor. Cells cultured in medium alone were used as control (gray). Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 Human DCs cytokine induction upon LPS or CpG stimulation in the presence or blockade of BTK. Normalized levels of IL-6, IL-12, TNF-α, and IL10 mRNA from healthy human DCs (n = 10) upon LPS or CpG stimulation in the presence or absence of ibrutinib. Data are expressed as normalized fold expression levels with respect to cells not stimulated. Bar graphs show values of 2 (+ SD) technical replicates. P values were calculated by using unpaired Student t test (*P = ). ns, Not significant. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 STAT1/3 upregulation in human DCs upon LPS or CpG stimulation in the presence or blockade of BTK. DCs from 10 healthy donors were stimulated with LPS or CpG in the presence or absence of Ibrutinib. STAT1 and STAT3 mRNA levels were measured and normalized to cells not stimulated. Each datum represents the mean of 2 (+ SD) independent repeat. P values were calculated by using unpaired Student t test (**P = ). ns, Not significant. Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig E1 Human DCs activation from healthy control (BTK+/+) (left panel) and XLA patient (BTK−/−) (right panel) upon LPS or CpG stimulation in terms of CD86, CD83, CD80, and HLA-DR upregulation (representative experiment) (density plot). Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Fig E2 Human DCs activation from healthy donors without and with ibrutinib upon LPS or CpG stimulation in terms of CD86, CD83, CD80, and HLA-DR upregulation (representative experiment) (density plot). Journal of Allergy and Clinical Immunology  , e4DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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