Presentation is loading. Please wait.

Presentation is loading. Please wait.

Targeting allergen to FcγRI reveals a novel TH2 regulatory pathway linked to thymic stromal lymphopoietin receptor  Kathryn E. Hulse, PhD, Amanda J. Reefer,

Published by镇恩 夏 Modified over 5 years ago

Similar presentations

Presentation on theme: "Targeting allergen to FcγRI reveals a novel TH2 regulatory pathway linked to thymic stromal lymphopoietin receptor  Kathryn E. Hulse, PhD, Amanda J. Reefer,"— Presentation transcript:

1 Targeting allergen to FcγRI reveals a novel TH2 regulatory pathway linked to thymic stromal lymphopoietin receptor  Kathryn E. Hulse, PhD, Amanda J. Reefer, MS, Victor H. Engelhard, PhD, James T. Patrie, MS, Steven F. Ziegler, PhD, Martin D. Chapman, PhD, Judith A. Woodfolk, MBChB, PhD  Journal of Allergy and Clinical Immunology  Volume 125, Issue 1, Pages e8 (January 2010) DOI: /j.jaci Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 The TH2-promoting effect of TSLP is weak in atopic subjects. CD4+ T cells were cocultured with moDCs primed for 48 hours with TSLP. A, Cells were stained for intracellular cytokines (day 10) and analyzed by means of flow cytometry, gating on CD4+ T cells. Relative change for each T-cell type compared with nonstimulated cells is expressed as NI (n = 5 subjects per group). Error bars represent 95% CIs. B, Representative dot plots showing nonstimulated (NS) and TSLP-stimulated CD4+ T cells stained for IL-4 and IFN-γ. Percentages of cells in each quadrant are shown. C, Secreted cytokines in nonstimulated (NS) and TSLP-stimulated cultures (n = 5 subjects per group). Bars represent means ± SEMs. ∗Significant versus control group. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Dendritic cells coprimed with H22–Fel d 1 and TSLP provide a potent TH2 stimulus in atopic subjects. CD4+ T cells were cocultured with moDCs that had been primed for 48 hours with Fel d 1 (Fel), H22–Fel d 1 (H-Fel), TSLP, or H22–Fel d 1 plus TSLP. Cytokine-positive T cells were analyzed on day 10. A, Summary data showing the percentage of each T-cell type induced by different stimuli (n = 5 subjects per group). Values are geometric means. Black and gray asterisks denote significant differences for IL-10+ and IL-4+ cell types, respectively, after adjusting for multiple comparisons. B, Relative change in each T-cell type for H22–Fel d 1 plus TSLP versus H22–Fel d 1 alone (left) and H22–Fel d 1 plus TSLP versus TSLP alone (right; n = 5 per group). Error bars represent 95% CIs. ∗Significant after adjusting for multiple comparisons. C, Representative dot plot from a patients with AD showing the phenotype of gated CD4+ T cells induced by different stimuli. NS, Nonstimulated. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 The TH2-promoting effect of coprimed moDCs is OX40L-independent. A, Expression of OX40L on moDCs after priming with different stimuli for 48 hours. Bars represent the mean fluorescence intensity (MFI) ± SEM (n = 5 subjects per group). ∗P < .05. B, Effect of anti-OX40L mAb on the percentage of IL-4+ T cells induced (day 10) by atopic moDCs primed with different stimuli for 48 hours. Bars represent the means ± SEMs (n = 4 subjects). C, CCL17/TARC levels in supernatants from cultures containing moDCs primed with different stimuli for 48 hours. Bars represent the means ± SEMs (n = 5 subjects per group). Fel, Fel d 1; H-Fel, H22–Fel d 1; NS, nonstimulated. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 H22–Fel d 1 enhances TSLPr expression on atopic moDCs. A, Comparison of TSLPr expression on moDCs from atopic and nonatopic subjects. moDCs were primed with different stimuli, and expression of TSLPr was analyzed at 24 hours. Data are representative of 4 atopic and 4 nonatopic subjects. FSC, Forward scatter. B, Time course of TSLPr expression on moDCs (representative of 6 subjects). Solid red histogram corresponds to the fluorescence-minus-one control (FMO). C, TSLPr expression on blood DCs. DCs were isolated, primed with different stimuli, and analyzed for TSLPr expression at 24 hours. Data are representative of 3 experiments. Fel, Fel d 1; H-Fel, H22–Fel d 1; NS, nonstimulated. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 FcR signaling components modulate H22–Fel d 1–induced TSLP receptor expression. A, Calcium mobilization in atopic moDCs primed with different stimuli. The y-axis represents the ratio of free to bound calcium. Data are representative of 4 atopic subjects. B, Effect of kinase inhibitors on H22–Fel d 1–induced TSLPr expression in atopic DCs. Cells were incubated with inhibitors for 1 hour before priming with H22–Fel d 1 and analyzing for TSLPr expression (24 hours). Inhibitors tested were PP2 (SRTK inhibitor), piceatannol (Syk kinase inhibitor), and LY (PI3-kinase inhibitor). Data are shown for 8 different atopic patients. ∗P ≤ .05. Fel, Fel d 1; H-Fel, H22–Fel d 1. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 Blocking TSLPr upregulation in coprimed moDCs abolishes their TH2-promoting effect. Atopic moDCs were incubated with no inhibitor (No inh.) or with LY before priming with different stimuli. Cells were then washed and cultured with CD4+ T cells for 7 days. Intracellular cytokine expression was analyzed by flow cytometry. Dot plots show the phenotype of gated CD4+ T cells. H-Fel, H22–Fel d 1; NS, nonstimulated. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Atopic DCs coprimed with H22–Fel d 1 and TSLP induce enhanced secretion of TH2 cytokines. MoDCs isolated from subjects with cat allergy with and without AD were cocultured with CD4+ T cells after priming for 48 hours with Fel d 1 (Fel), H22–Fel d 1 (H-Fel), TSLP, or H22–Fel d 1 plus TSLP. Secreted cytokines were measured on day 10. Bars represent means ± SEMs (n = 5 per group). ∗Significant after adjusting for multiple comparisons. NS, Nonstimulated. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Representative data showing the effect of anti-OX40L antibody on T-cell responses induced by atopic moDCs. CD4+ T cells were cocultured in the presence or absence of anti-OX40L mAb with moDCs that had been primed for 48 hours with different stimuli. Dot plots show the phenotype of gated CD4+ T cells in the presence or absence of anti-OX40L mAb or isotype control. H-Fel, H22–Fel d 1; NS, nonstimulated. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 Analysis of the neutralizing capacity of anti-OX40L antibody
Analysis of the neutralizing capacity of anti-OX40L antibody. Blood DCs were primed with TSLP for 48 hours before coculture with CD4+ T cells in the presence or absence of anti-OX40L mAb (7 days). A, Frequency of IL-4+ T cells detected in the presence or absence of antibody (Ab). An isotype control had no effect (data not shown). Histogram shows OX40 expression on IL-4+ T cells versus IL-4–negative cells. B, Comparison of OX40L expression on nonstimulated (NS) and TSLP-primed DCs (48 hours). Data are representative of 2 experiments. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

11 Change in the percentage of TSLPr+ cells induced by different stimuli in atopic moDCs. MoDCs were primed with different stimuli, and expression of TSLPr was analyzed at 24 hours. ∗P < .05 and ∗∗P < .01 (n = 4 atopic subjects). Fel, Fel d 1; H-Fel, H22–Fel d 1; NS, nonstimulated. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

12 Analysis of the effect of human serum on TSLPr expression
Analysis of the effect of human serum on TSLPr expression. THP-1 monocytes were primed with H22–Fel d 1 in the presence of FBS or atopic and nonatopic serum, and TSLPr expression was measured at 24 hours. Data are representative of 8 experiments. H-Fel, H22–Fel d 1; FMO, fluorescence-minus-one control. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

13 TSLPr is not induced by low-dose LPS
TSLPr is not induced by low-dose LPS. Cells were primed with H22–Fel d 1 or LPS (0.5 ng/mL or 1 μg/mL), and TSLPr expression was analyzed by flow cytometry (mean ± SD for 7 subjects). ∗P < .05. H-Fel, H22–Fel d 1; MFI, mean fluorescent intensity; NS, nonstimulated. Journal of Allergy and Clinical Immunology  , e8DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Download ppt "Targeting allergen to FcγRI reveals a novel TH2 regulatory pathway linked to thymic stromal lymphopoietin receptor  Kathryn E. Hulse, PhD, Amanda J. Reefer,"

Similar presentations

Multiple-checkpoint inhibition of thymic stromal lymphopoietin–induced TH2 response by TH17-related cytokines  Sofia I. Bogiatzi, BSc, Maude Guillot-Delost,

Multiple-checkpoint inhibition of thymic stromal lymphopoietin–induced TH2 response by TH17-related cytokines  Sofia I. Bogiatzi, BSc, Maude Guillot-Delost,
Dorothea Dijkstra, MSc, Rob Leurs, PhD, Paul Chazot, PhD, Fiona C

Dorothea Dijkstra, MSc, Rob Leurs, PhD, Paul Chazot, PhD, Fiona C
Human slan (6-sulfo LacNAc) dendritic cells are inflammatory dermal dendritic cells in psoriasis and drive strong Th17/Th1 T-cell responses  Anja Hänsel,

Human slan (6-sulfo LacNAc) dendritic cells are inflammatory dermal dendritic cells in psoriasis and drive strong Th17/Th1 T-cell responses  Anja Hänsel,
Flow cytometry imaging identifies rare TH2 cells expressing thymic stromal lymphopoietin receptor in a “proallergic” milieu  Amanda J. Reefer, MS, Kathryn.

Flow cytometry imaging identifies rare TH2 cells expressing thymic stromal lymphopoietin receptor in a “proallergic” milieu  Amanda J. Reefer, MS, Kathryn.
Correlation of allergen-specific T follicular helper cell counts with specific IgE levels and efficacy of allergen immunotherapy  Yin Yao, MD, Cai-Ling.

Correlation of allergen-specific T follicular helper cell counts with specific IgE levels and efficacy of allergen immunotherapy  Yin Yao, MD, Cai-Ling.
Induction of anergic allergen-specific suppressor T cells using tolerogenic dendritic cells derived from children with allergies to house dust mites 

Induction of anergic allergen-specific suppressor T cells using tolerogenic dendritic cells derived from children with allergies to house dust mites 
Mechanical injury polarizes skin dendritic cells to elicit a TH2 response by inducing cutaneous thymic stromal lymphopoietin expression  Michiko K. Oyoshi,

Mechanical injury polarizes skin dendritic cells to elicit a TH2 response by inducing cutaneous thymic stromal lymphopoietin expression  Michiko K. Oyoshi,
Cell-to-cell contact between activated CD4+ T lymphocytes and unprimed monocytes interferes with a TH1 response  Miriam Wittmann, MD, Mareike Alter, Tanja.

Cell-to-cell contact between activated CD4+ T lymphocytes and unprimed monocytes interferes with a TH1 response  Miriam Wittmann, MD, Mareike Alter, Tanja.
IgE cross-linking impairs monocyte antiviral responses and inhibits influenza-driven TH1 differentiation  Regina K. Rowe, MD, PhD, David M. Pyle, MD,

IgE cross-linking impairs monocyte antiviral responses and inhibits influenza-driven TH1 differentiation  Regina K. Rowe, MD, PhD, David M. Pyle, MD,
Reduced IFN-γ receptor expression and attenuated IFN-γ response by dendritic cells in patients with atopic dermatitis  Eva Gros, MSc, Susanne Petzold,

Reduced IFN-γ receptor expression and attenuated IFN-γ response by dendritic cells in patients with atopic dermatitis  Eva Gros, MSc, Susanne Petzold,
Human B cells promote T-cell plasticity to optimize antibody response by inducing coexpression of TH1/TFH signatures  Jelle de Wit, PhD, Tineke Jorritsma,

Human B cells promote T-cell plasticity to optimize antibody response by inducing coexpression of TH1/TFH signatures  Jelle de Wit, PhD, Tineke Jorritsma,
Toll-like receptor 7–induced naive human B-cell differentiation and immunoglobulin production  Mark C. Glaum, MD, PhD, Shilpi Narula, MD, Decheng Song,

Toll-like receptor 7–induced naive human B-cell differentiation and immunoglobulin production  Mark C. Glaum, MD, PhD, Shilpi Narula, MD, Decheng Song,
Ting-ting Zhang, MSc, Klaus Okkenhaug, PhD, Baher F

Ting-ting Zhang, MSc, Klaus Okkenhaug, PhD, Baher F
Bruton tyrosine kinase mediates TLR9-dependent human dendritic cell activation  Vassilios Lougaris, MD, Manuela Baronio, PhD, Massimiliano Vitali, PhD,

Bruton tyrosine kinase mediates TLR9-dependent human dendritic cell activation  Vassilios Lougaris, MD, Manuela Baronio, PhD, Massimiliano Vitali, PhD,
Ramses Ilarraza, PhD, Yingqi Wu, Christopher D

Ramses Ilarraza, PhD, Yingqi Wu, Christopher D
Targeting Toll-like receptors on dendritic cells modifies the TH2 response to peanut allergens in vitro  Pierre Pochard, PhD, Brian Vickery, MD, M. Cecilia.

Targeting Toll-like receptors on dendritic cells modifies the TH2 response to peanut allergens in vitro  Pierre Pochard, PhD, Brian Vickery, MD, M. Cecilia.
Responsiveness to respiratory syncytial virus in neonates is mediated through thymic stromal lymphopoietin and OX40 ligand  Junyan Han, PhD, Azzeddine.

Responsiveness to respiratory syncytial virus in neonates is mediated through thymic stromal lymphopoietin and OX40 ligand  Junyan Han, PhD, Azzeddine.
Triggering of specific Toll-like receptors and proinflammatory cytokines breaks allergen- specific T-cell tolerance in human tonsils and peripheral blood 

Triggering of specific Toll-like receptors and proinflammatory cytokines breaks allergen- specific T-cell tolerance in human tonsils and peripheral blood 
Dawn C. Newcomb, PhD, Madison G

Dawn C. Newcomb, PhD, Madison G
Volume 141, Issue 3, Pages (September 2011)

Volume 141, Issue 3, Pages (September 2011)

Similar presentations

Ads by Google