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Human mast cells are major IL-22 producers in patients with psoriasis and atopic dermatitis  Shunya Mashiko, BPharm, Salim Bouguermouh, MD, PhD, Manuel.

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Presentation on theme: "Human mast cells are major IL-22 producers in patients with psoriasis and atopic dermatitis  Shunya Mashiko, BPharm, Salim Bouguermouh, MD, PhD, Manuel."— Presentation transcript:

1 Human mast cells are major IL-22 producers in patients with psoriasis and atopic dermatitis 
Shunya Mashiko, BPharm, Salim Bouguermouh, MD, PhD, Manuel Rubio, BSc, Nobuyasu Baba, PhD, Robert Bissonnette, MD, Marika Sarfati, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 136, Issue 2, Pages e1 (August 2015) DOI: /j.jaci Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Distribution of immune cells in skin cell suspensions freshly isolated from skin biopsy specimens of patients with psoriasis. Biopsies were performed on plaques (L; 22 patients) and nonlesional skin (NL; 7 patients) of patients with psoriasis. A, Viable cell count using trypan blue dye exclusion. B, Gating strategy on Aqua LIVE/DEAD negative cells in relation to cell size (FSC). C, Gating strategy on viable CD45+ hematopoietic cells (left panel), CD45+ cells (percentage; middle panel), and (absolute number; right panel). D, CD45+CD3+ and CD3− cells (percentage). E, CD45+CD14+ versus c-Kit+ cells (percentage). Fig 1, A, C, and E: unpaired t test (Mann-Whitney test). Fig 1, D, Wilcoxon test. ***P < .001 and ****P < Journal of Allergy and Clinical Immunology  , e1DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 c-Kit+ cells are major IL-22 producers, and CD3+ T cells are the main source of IL-17 in psoriatic plaques. A and B, Intracytoplasmic staining of skin cell suspensions using anti–IL-22 mAb, anti–IL-17A mAb, or isotype-matched control mAb after gating on CD45+ cells: Fig 2, A, CD45+IL-22+ cells (percentage); Fig 2, B, CD45+IL-17+ cells (percentage). C, IL-22 and IL-17 expression gated on c-Kit+CD3− and c-Kit−CD3+ cells. D, CD3+ compared with c-Kit+IL-22+CD45+ cells (percentage). E, CD3+ versus c-Kit+IL-17+CD45+ cells (percentage). Data show 1 representative experiment of 5. **P < .01 and ****P < Journal of Allergy and Clinical Immunology  , e1DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Mast cells (c-kit+FcεRI+ cells) produce IL-22 and IL-17 and express IL-17R in psoriatic plaques. A, Intracytoplasmic IL-22 and IL-17 production in relation to CD127 expression after gating on c-Kit+FcεRI+ mast cells. Shown is 1 representative experiment of 10. B, Morphology of fluorescence-activated cell sorting–sorted c-Kit+FcεRI+ mast cells and c-Kit−FcεRI−CD3+ T cells after gating on CD45+ hematopoietic viable cells. C, Cell-surface versus intracytoplasmic staining with anti–IL-22 or anti–IL-17 mAb in relation to c-Kit expression after gating on CD45+ cells. D, IL-17RA and c-Kit cell-surface expression after gating on c-Kit+FcεRI+ and c-Kit−FcεRI−CD3+ cells. Fig 3, B-D, Data show 1 representative experiment of 2 to 3. Journal of Allergy and Clinical Immunology  , e1DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Mast cells (c-kit+FcεRI+ cells) are a major source of IL-22 in patients with atopic dermatitis. Skin and colonic biopsy specimens were collected from patients with atopic dermatitis (AD; n = 6), patients with psoriasis (PS; n = 8), and patients with Crohn disease (CD; n = 2). A, IL-22 intracytoplasmic staining with anti–IL-22 mAb or isotype-matched control mAb after gating on mast cells (c-Kit+FcεRI+). B, CD3+ and c-Kit+IL-22+ CD45+ cells (percentage). C, Frequency of IL-22+ cells among mast cells in patients with psoriasis, patients with atopic dermatitis, and patients with Crohn disease. Journal of Allergy and Clinical Immunology  , e1DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 Quantification of IL-22 and IL-17 mRNA expression at the single-cell level in human peripheral blood memory CD4+ T cells. Human peripheral blood memory CD4+ T cells were activated for 5 to 7 days with anti-CD3 (10 μg/mL)–coated 24-well plates. A, IL-22 and IL-17 intracytoplasmic staining before and after PMA/ionomycin stimulation. Top panel, Isotype-matched control mAb on stimulated cells. B, IL-22 and IL-17 expression of mRNA as in Fig 5, A. C, Activated human memory T cells were mixed with murine cells at a 1:1 ratio. β2-Microglobulin mRNA expression in relation to human CD45 cell-surface expression (top panel) and IL-22 and IL-17 and β2-microglobulin mRNA coexpression are shown. Data show 1 representative experiment of 2. Journal of Allergy and Clinical Immunology  , e1DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 Quantification of IL-22 and IL-17 mRNA expression at the single-cell level in human mast cells and T cells in skin biopsy specimens of patients with psoriasis. β2-Microglobulin, IL-22, and IL-17 mRNA expression after gating on mast cells (c-Kit+FcεRI+CD3−) and T cells (c-Kit−FcεRI−CD3+) in human psoriatic skin biopsy specimens. Data show 1 representative experiment of 3. Journal of Allergy and Clinical Immunology  , e1DOI: ( /j.jaci ) Copyright © 2015 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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