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The mannose receptor negatively modulates the Toll-like receptor 4–aryl hydrocarbon receptor–indoleamine 2,3-dioxygenase axis in dendritic cells affecting.

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Presentation on theme: "The mannose receptor negatively modulates the Toll-like receptor 4–aryl hydrocarbon receptor–indoleamine 2,3-dioxygenase axis in dendritic cells affecting."— Presentation transcript:

1 The mannose receptor negatively modulates the Toll-like receptor 4–aryl hydrocarbon receptor–indoleamine 2,3-dioxygenase axis in dendritic cells affecting T helper cell polarization  Fabián Salazar, MSc, Laurence Hall, BSc, Ola H. Negm, PhD, Dennis Awuah, MSc, Patrick J. Tighe, PhD, Farouk Shakib, PhD, FRCPath, Amir M. Ghaemmaghami, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 137, Issue 6, Pages e2 (June 2016) DOI: /j.jaci Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 A, IDO activity in DCs stimulated with LPS (n = 5). B, IDO activity in DCs stimulated with ultrapure LPS (n = 2). C, IDO activity in myeloid DCs stimulated with 0.01 μg/mL LPS (n = 2). D, IDO1/IDO2 gene expression in DCs stimulated with 0.01 μg/mL LPS (n ≥ 3). E, IDO1 protein expression in DCs stimulated with 0.1 μg/mL LPS (n = 5). All stimulations are for 24 hours. **P ≤ .01 and ****P ≤ .0001. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 A, IDO activity in DCs stimulated for 24 hours with allergen extracts (10 μg/mL) and LPS (0.01 μg/mL; n ≥ 4). BGP, Bermuda grass pollen; GC, German cockroach. B, IDO activity in DCs stimulated for 24 hours with mannan (M; 10 μg/mL) and LPS (n = 5). C, IDO1/IDO2 gene expression in DCs stimulated for 24 hours with mannan and LPS (n = 3). In costimulation experiments cells were stimulated with LPS, followed immediately by allergen extracts. **P ≤ .01, ***P ≤ .001, and ****P ≤ .0001. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 A, Cytokine production by DCs stimulated for 24 hours with mannan (M) and LPS. B, IL-12 production by DCs stimulated with allergen extracts and LPS. BGP, Bermuda grass pollen; GC, German cockroach. C, DC phenotype after 24 hours of stimulation with mannan and LPS. D and E, Percentage of IFN-γ–positive cells and IFN-γ production by T cells in autologous cocultures, respectively. F, IL-4 and IFN-γ intracellular staining on CD4+ cells (n = 3). *P ≤ .05, **P ≤ .01, and ***P ≤ .001. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 A, IDO activity in CT-DCs and MRlow-DCs stimulated with mannan (M) and LPS for 24 hours. B, MR and IDO1 protein expression in DCs stimulated with mannan and LPS for 24 hours. Median fluorescence intensity (MFI) values were normalized by using a control unstimulated sample (n = 3). *P ≤ .05, **P ≤ .01, and ****P ≤ .0001. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 A, IDO activity in CT-DCs and AhRlow-DCs stimulated or not with LPS for 24 hours. B, AHR and CYP1A1 gene expression in DCs stimulated or not with mannan (M) and LPS for 24 hours. C, IL-12p70 and IL-10 production by CT-DCs and AhRlow-DCs stimulated or not with LPS for 24 hours (n = 3). **P < .01 and ****P ≤ .0001. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 A, Heat maps showing the relative abundance of proteins upstream of the NF-κB pathway by using DCs stimulated with mannan (M) and LPS. B, Protein expression of phospho-p65, NIK, and RelB. Data are shown as geometric means (n = 3). C, Protein expression of RelB (90 minutes), NIK (90 minutes), phospho-p65 (18 hours), and TRAF3 (18 hours). D, RELB gene expression in DCs stimulated with mannan and LPS for 24 hours (n = 2). *P ≤ .05 and ****P ≤ .0001. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig 7 MR-mediated IDO downregulation in human DCs involved the transcription factors AhR and RelB, which have an effect on T helper cell polarization. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Fig E1 A, Quantitative RT-PCR analysis of MR gene expression in CT-DCs and MRlow-DCs (n = 2). B, Flow cytometric analysis of MR protein expression in CT-DCs and MRlow-DCs. Median fluorescence intensity (MFI) is shown (n = 5). C, Flow cytometric analysis of DC-SIGN, HLA-DR, TLR4, CD14, myeloid differentiation protein-2 (MD-2) and AhR protein expression. Median fluorescence intensity (MFI) is shown (n ≥ 2). **P < .01 and ****P ≤ .0001. Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 Fig E2 A, Quantitative RT-PCR analysis of AHR gene expression in CT-DCs and AhRlow-DCs (n = 2). B, Flow cytometric analysis of AhR protein expression in CT-DCs and AhRlow-DCs. Median fluorescence intensity (MFI) is shown (n ≥ 3). C, Flow cytometric analysis of MR, DC-SIGN, TLR4, CD14, myeloid differentiation protein-2 (MD-2), and CD86 protein expression. Median fluorescence intensity (MFI) is shown (n ≥ 2). ***P < .001 and ****P < Journal of Allergy and Clinical Immunology  , e2DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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