Presentation is loading. Please wait.

Presentation is loading. Please wait.

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

Published byNickolas Cameron Modified over 5 years ago

Similar presentations

Presentation on theme: "Targeting Toll-like receptors on dendritic cells modifies the TH2 response to peanut allergens in vitro  Pierre Pochard, PhD, Brian Vickery, MD, M. Cecilia."— Presentation transcript:

1 Targeting Toll-like receptors on dendritic cells modifies the TH2 response to peanut allergens in vitro  Pierre Pochard, PhD, Brian Vickery, MD, M. Cecilia Berin, PhD, Alexander Grishin, PhD, Hugh A. Sampson, MD, Michael Caplan, MD, PhD, Kim Bottomly, PhD  Journal of Allergy and Clinical Immunology  Volume 126, Issue 1, Pages e5 (July 2010) DOI: /j.jaci Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 DC treatment with HKE alters T-cell immune responses to peanut. CD4+ T cells were cocultured (ratio 10:1) with CPE-pulsed BMDCs stimulated with CPE (50 μg/mL), HKE (1 bacterium per BMDC), or both for 72 hours. A, Concentrations in IL-4, IL-5, IL-13, and IFN-γ were determined. Concentrations represent means ± SEMs (n = 5). ∗P < .05. B, After 5 days, proliferation of CD3+ cells was analyzed by means of flow cytometry. One representative experiment of 5 is shown. CFSE, Carboxyfluorescein succinimidyl ester; CM, complete media. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 HKE alters peanut response by triggering TLR signaling in DCs. Wild-type (WT) CD4+ T cells were cocultured (ratio 10:1) with CPE-pulsed BMDCs (WT vs MyD88/TRIF-KO) stimulated with CPE (50 μg/mL), HKE (1 bacterium per BMDC), or both for 72 hours. Concentrations of IL-4, IL-13, and IFN-γ were determined. Concentrations represent means ± SEMs (n = 5). After 5 days, proliferation of CD3+ cells was analyzed by means of flow cytometry. Percentages of proliferative cells are indicated by means ± SEMs (n = 5). Statistical differences are indicated as follows: ∗∗P < .01. CM, Complete media. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 HKE has a long-lasting effect on peanut response. CD4+ T cells were cocultured (ratio 10:1) with BMDCs stimulated with CPE (50 μg/mL), HKE (1 bacterium per BMDC), or both. After 7 days, they were washed, counted, and cocultured with CPE-pulsed BMDCs for 72 hours. A, Supernatants were analyzed for the presence of IL-5, IL-13, IL-17, and IFN-γ. ∗P < .05, ∗∗P < .01. B, Proliferation of CD3+ cells was determined by means of flow cytometry. One representative experiment of 5 is shown. CFSE, Carboxyfluorescein succinimidyl ester; CM, complete media. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 HKE changes peanut-induced cytokine production by BMDCs. BMDCs were stimulated either with CPE (50 μg/mL), HKE (ratio of 1 bacterium per BMDC), or both stimuli for 24 hours. Unstimulated BMDCs were used as controls. Supernatants were analyzed by means of ELISA for production of IL-10, IL-12, IL-6, and TNF-α. Concentrations represent means ± SEMs (n = 15). ∗P < .05, ∗∗P < .01. CM, Complete media. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 Adding TLR4 and TLR9 ligands to CPE stimulation synergizes IL-10 production, whereas only TLR9 leads to IL-12 synergy by BMDCs. BMDCs (wild-type [WT] vs TLR4-knockout [KO] or TLR9-KO) were either stimulated with CPE (50 μg/mL), ultrapure LPS (500 ng/mL), ultrapure CpG (500 ng/mL), or a combination of both stimuli for 24 hours. Supernatants were analyzed by means of ELISA for production of IL-10 and IL-12. Concentrations represent means ± SEMs (n = 4). ∗∗P ≤ .01. CM, Complete media. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 E coli partially inhibits the peanut-induced immune response through IL-12 production. CD4+ T cells were cocultured (ratio 10:1) with CPE-pulsed BMDCs stimulated with CPE (50 μg/mL), HKE (1 bacterium per BMDC), or both for 72 hours. BMDCs were preincubated with either an anti–IL-12 antibody or isotype control at 20 μg/mL. The anti–IFN-γ antibody (20 μg/mL) was added with T cells. Antibody concentrations were maintained during the whole time of the coculture. Concentrations of IL-4, IL-5, and IL-13 were determined, as well as concentrations of IFN-γ and IL-10. Concentrations represent means ± SEMs. ∗∗P ≤ .01). After 5 days, proliferation of CD3+ cells was analyzed by means of flow cytometry. Four combined experiments involving at least 5 mice are shown. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Stimulation of DCs with peanut allergens induces TLR-independent IL-17 secretion. CD4+ T cells were cocultured (ratio 10:1) with CPE-pulsed BMDCs stimulated with CPE (50 μg/mL), HKE (1 bacterium per BMDC), or both for 72 hours. A, Wild-type (WT) BMDCs and T cells were used. Concentrations of IL-17 were determined. Concentrations represent means ± SEMs (n = 5). ∗P < .05, ∗∗P ≤ .01). B, MyD88/TRIF double-knockout (KO) BMDCs and WT T cells were used. IL-17 concentration was measured. Concentrations represent means ± SEMs (n = 5). ∗P < .05, ∗∗P ≤ .01). CM, Complete media. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 HKE alters peanut response by triggering TLR cell signaling
HKE alters peanut response by triggering TLR cell signaling. Wild-type (WT) CD4+ T cells were cocultured (ratio 10:1) with CPE-pulsed BMDCs (WT vs MyD88/TRIF-knockout [KO]) stimulated with CPE (50 μg/mL), HKE (1 bacterium per BMDC), or both for 72 hours. After 5 days, proliferation of CD3+ cells was analyzed by means of flow cytometry. One representative experiment of 5 is shown. CFSE, Carboxyfluorescein succinimidyl ester; CM, complete media. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 HKE enhances CPE-induced BMDC maturation
HKE enhances CPE-induced BMDC maturation. Wild-type (WT) and MyD88-knockout BMDCs were stimulated with CPE (50 μg/mL) in the presence or absence of HKE (ratio of 1 bacterium per BMDC) or LPS (500 ng/mL) for 24 hours. BMDCs were analyzed for CD80, CD86, CD40, and IAb by means of flow cytometry. Solid histograms represent unstimulated BMDCs. One representative experiment of 5 is shown. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

11 Adding TLR4 or TLR9 ligands to CPE stimulation increases IL-6 and TNF-α production by BMDCs. Wild-type (WT), TLR4-knockout (KO), and TLR9-KO BMDCs were either stimulated with CPE (50 μg/mL), ultrapure LPS (500 ng/mL), ultrapure CpG (500 ng/mL), or a combination of both stimuli for 24 hours. Supernatants were analyzed by means of ELISA for production of TNF-α and IL-6. Concentrations represent means ± SEMs (n = 4). ∗∗P ≤ .01. CM, Complete media. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

12 IL-10 is not responsible for HKE's effect on the CPE-induced immune response. CD4+ T cells (from IL-10–knockout mice) were cocultured (ratio 10:1) with CPE-pulsed BMDCs (from IL-10–knockout mice) stimulated with CPE (50 μg/mL), HKE (1 bacterium per BMDC), or both for 72 hours. A, Concentrations of IL-4, IL-5, IL-13, and IFN-γ were determined. Concentrations represent means ± SEMs (n = 5). ∗P < .05. B, After 5 days, proliferation of CD3+ cells was analyzed by means of flow cytometry. One representative experiment of 5 is shown. CFSE, Carboxyfluorescein succinimidyl ester; CM, complete media. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2010 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Download ppt "Targeting Toll-like receptors on dendritic cells modifies the TH2 response to peanut allergens in vitro  Pierre Pochard, PhD, Brian Vickery, MD, M. Cecilia."

Similar presentations

Maria B. Sukkar, PhD, Shaoping Xie, PhD, Nadia M

Maria B. Sukkar, PhD, Shaoping Xie, PhD, Nadia M
Toll-like receptor 2 ligands promote chronic atopic dermatitis through IL-4–mediated suppression of IL-10  Susanne Kaesler, PhD, Thomas Volz, MD, Yuliya.

Toll-like receptor 2 ligands promote chronic atopic dermatitis through IL-4–mediated suppression of IL-10  Susanne Kaesler, PhD, Thomas Volz, MD, Yuliya.
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 
Immunomodulatory effects of aqueous birch pollen extracts and phytoprostanes on primary immune responses in vivo  Jan Gutermuth, MD, Mayte Bewersdorff,

Immunomodulatory effects of aqueous birch pollen extracts and phytoprostanes on primary immune responses in vivo  Jan Gutermuth, MD, Mayte Bewersdorff,
Nonpathogenic Bacteria Alleviating Atopic Dermatitis Inflammation Induce IL-10- Producing Dendritic Cells and Regulatory Tr1 Cells  Thomas Volz, Yuliya.

Nonpathogenic Bacteria Alleviating Atopic Dermatitis Inflammation Induce IL-10- Producing Dendritic Cells and Regulatory Tr1 Cells  Thomas Volz, Yuliya.
Therapeutic reversal of food allergen sensitivity by mature retinoic acid–differentiated dendritic cell induction of LAG3+CD49b−Foxp3− regulatory T cells 

Therapeutic reversal of food allergen sensitivity by mature retinoic acid–differentiated dendritic cell induction of LAG3+CD49b−Foxp3− regulatory T cells 
Histamine receptor 2 modifies dendritic cell responses to microbial ligands  Remo Frei, PhD, Ruth Ferstl, PhD, Patrycja Konieczna, MSc, Mario Ziegler,

Histamine receptor 2 modifies dendritic cell responses to microbial ligands  Remo Frei, PhD, Ruth Ferstl, PhD, Patrycja Konieczna, MSc, Mario Ziegler,
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 
Human TH17 cell development requires processing of dendritic cell–derived CXCL8 by neutrophil elastase  Yuri Souwer, PhD, Tom Groot Kormelink, PhD, Esther.

Human TH17 cell development requires processing of dendritic cell–derived CXCL8 by neutrophil elastase  Yuri Souwer, PhD, Tom Groot Kormelink, PhD, Esther.
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,
Differential cytokine induction by the human skin–associated autoallergen thioredoxin in sensitized patients with atopic dermatitis and healthy control.

Differential cytokine induction by the human skin–associated autoallergen thioredoxin in sensitized patients with atopic dermatitis and healthy control.
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,
Interferon response factor 3 is essential for house dust mite–induced airway allergy  Thomas Marichal, DVM, Denis Bedoret, DVM, PhD, Claire Mesnil, DVM,

Interferon response factor 3 is essential for house dust mite–induced airway allergy  Thomas Marichal, DVM, Denis Bedoret, DVM, PhD, Claire Mesnil, DVM,
Ting-ting Zhang, MSc, Klaus Okkenhaug, PhD, Baher F

Ting-ting Zhang, MSc, Klaus Okkenhaug, PhD, Baher F
Ramses Ilarraza, PhD, Yingqi Wu, Christopher D

Ramses Ilarraza, PhD, Yingqi Wu, Christopher D
Glycation of a food allergen by the Maillard reaction enhances its T-cell immunogenicity: Role of macrophage scavenger receptor class A type I and II 

Glycation of a food allergen by the Maillard reaction enhances its T-cell immunogenicity: Role of macrophage scavenger receptor class A type I and II 
Lung dendritic cells induce TH17 cells that produce TH2 cytokines, express GATA-3, and promote airway inflammation  Marianne Raymond, PhD, Vu Quang Van,

Lung dendritic cells induce TH17 cells that produce TH2 cytokines, express GATA-3, and promote airway inflammation  Marianne Raymond, PhD, Vu Quang Van,
Glucocorticoids promote intrinsic human TH17 differentiation

Glucocorticoids promote intrinsic human TH17 differentiation
Basophil-derived IL-4 promotes epicutaneous antigen sensitization concomitant with the development of food allergy  Maryam Hussain, MSc, Loïc Borcard,

Basophil-derived IL-4 promotes epicutaneous antigen sensitization concomitant with the development of food allergy  Maryam Hussain, MSc, Loïc Borcard,

Similar presentations

Ads by Google