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A thymic stromal lymphopoietin–responsive dendritic cell subset mediates allergic responses in the upper airway mucosa  Guro R. Melum, MD, Lorant Farkas,

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Presentation on theme: "A thymic stromal lymphopoietin–responsive dendritic cell subset mediates allergic responses in the upper airway mucosa  Guro R. Melum, MD, Lorant Farkas,"— Presentation transcript:

1 A thymic stromal lymphopoietin–responsive dendritic cell subset mediates allergic responses in the upper airway mucosa  Guro R. Melum, MD, Lorant Farkas, MD, PhD, Cecilie Scheel, MD, Brenda Van Dieren, MSc, Einar Gran, MD, Yong-Jun Liu, MD, PhD, Finn-Eirik Johansen, PhD, Frode L. Jahnsen, MD, PhD, Espen S. Baekkevold, PhD  Journal of Allergy and Clinical Immunology  Volume 134, Issue 3, Pages e7 (September 2014) DOI: /j.jaci Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Nasal mucosal CD1c+ DCs express functional TSLPR. A and B, TSLPR assessed by using fluorescence-activated cell sorting analysis in enzymatically digested biopsy specimens from nasal mucosa (Fig 1, A) and CD1c+ DCs bead sorted from PBMCs (Fig 1, B) that were freshly isolated, cultured overnight, or stimulated with poly I:C for 24 hours. C, pSTAT5 assessed by using fluorescence-activated cell sorting analysis in CD1c+ DCs from nasal mucosa (left) and blood (right) stimulated for 15 minutes with 300 ng/mL rhTSLP. All figures are representative of more than 3 donors. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Nasal mucosal CD1c+ DCs resemble CD1c+ DCs in peripheral blood but express a mature phenotype. A and B, Gating strategy (for all histograms in figure; Fig 2, A) and surface markers on nasal mucosal CD1c+ DCs (top) and blood-derived CD1c+ DCs (bottom; Fig 2, A and B). All figures are representative of more than 3 donors. FSC, Forward scatter; SSC, side scatter. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Nasal mucosal CD1a+CD1c+ DC counts increase in tissue after topical allergen challenge. A and B, CD1c+HLA-DR+ (Fig 3, A) and CD1c+CD1a+ (Fig 3, B) DCs in the epithelium and 250-μm subepithelial region are presented as cells per millimeter of basement membrane at days 0, 1, and 7 after allergen challenge in allergic (left) and nonallergic (right) subjects (n = 7). Significant P values are indicated, as calculated by using the Wilcoxon test. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Rapid induction of TSLP expression in the nasal mucosa by inflammatory TH2 cytokines. Nasal mucosal biopsy specimens were stimulated with inflammatory TH2 cytokines (IL-4, IL-13, and TNF-α) for 4 hours, and TSLP gene expression was assessed by means of RT-PCR. Gene expression of long-form TSLP (left) and short-form TSLP (right) is shown as relative expression to the housekeeping gene hypoxanthine phosphoribosyltransferase (HPRT; n = 7). Significant P values are indicated, as calculated by using the Wilcoxon test. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 TSLP induces CD1a expression and increases the migratory potential of TSLP-responding DCs. Blood-derived CD1c+ DCs were stimulated with TSLP, the TH2 cytokines IL-4 and IL-13, or both. A, Flow cytometric analysis of CD1a versus CCR7. The plot is representative of 6 donors. B and C, CD1a expression (Fig 5, B) and percentage of CCR7-expressing cells in CD1c+CD1a− cells (left) and CD1c+CD1a+ cells (right) after stimulation (Fig 5, C). Horizontal bars indicate means. Significant P values are indicated, as calculated by using the Wilcoxon test. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 TSLP enhances CD1c+ DC–mediated activation of allergen-specific TH2 cells. Blood-derived CD4+ T cells (105) were cultured together with an increasing number of autologous CD1c+ DCs × 103 (x-axis). T-cell proliferation is presented as the percentage of T cells in division measured by means of flow cytometry, and cytokine production is presented as picograms per milliliter of cytokine measured in culture medium for allergic (A) and healthy (B) donors. Values are presented as SEMs, and differences between groups were calculated by using 2-way ANOVA. P values represent the difference between grass pollen extract alone and grass pollen extract plus TSLP. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig E1 Immunohistochemical staining of antigen-presenting cell subsets in human nasal mucosal tissue. A-D, CD163 and CD11c (Fig E1, A), CD1c and CD11c (Fig E1, B), CD1c and CD163 (Fig E1, C), and CD1c and CD14 (Fig E1, D). The figure is representative of more than 7 donors (magnification ×400). Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Fig E2 Density of CD1c+ DCs, but not pDCs, correlates with symptom score after allergen challenge. A, Density of CD1c+ DCs in allergic donors versus total symptom score after 7 days of allergen challenge. The P value was calculated by using the Spearman correlation test. B, Density of pDCs in allergic donors versus total symptom score. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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