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UV Radiation Induces the Epidermal Recruitment of Dendritic Cells that Compensate for the Depletion of Langerhans Cells in Human Skin  Amine Achachi,

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Presentation on theme: "UV Radiation Induces the Epidermal Recruitment of Dendritic Cells that Compensate for the Depletion of Langerhans Cells in Human Skin  Amine Achachi,"— Presentation transcript:

1 UV Radiation Induces the Epidermal Recruitment of Dendritic Cells that Compensate for the Depletion of Langerhans Cells in Human Skin  Amine Achachi, Marc Vocanson, Philippe Bastien, Josette Péguet-Navarro, Sophie Grande, Catherine Goujon, Lionel Breton, Isabelle Castiel-Higounenc, Jean- François Nicolas, Audrey Gueniche  Journal of Investigative Dermatology  Volume 135, Issue 8, Pages (August 2015) DOI: /jid Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 UVR triggers the release of cytokines/chemokines and the afflux of CD45+ cells into the epidermis. (a) Design of the two clinical studies and collected skin samples. The upper panel shows the first study involving 27 human and suction blisters. Minimal erythema dose (MED) was determined at day-7, 1 hour after performing suction blisters on normal skin. The lower panel shows the second study involving 2 human subjects and skin biopsies. MED was determined at day-7. (b) Time-course release of IL-6, IL-8, IL-10, and tumor necrosis factor-α in the suction blister fluids from unexposed (control; white bars) and exposed skin areas (UV; black bars). The bars show the mean±SD cytokine levels (pg ml−1) from the 27 healthy volunteers. (c) Time-course quantification of the viable CD45+ cells in exposed (D+1–D+10) versus unexposed epidermis. Results are expressed as the mean±SD ratio of the percentage of viable CD45+ hematopoietic cells in exposed (UV-EC) versus unexposed (C-EC) epidermal cells from the 27 volunteers. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 Monocytes infiltrate the UV-exposed epidermis. (a) Epidermal cells were collected from unexposed (Control) or exposed (UV) skin samples and selected as a viable CD45+CD14+CD36+monocyte population using flow-cytometry gating. (b) Time-course analysis of monocyte recruitment in unexposed (Control) versus exposed (UV) epidermis. Data are expressed as percentage of cells in the viable 7AAD− CD45+ cell population from the 27 volunteers, and the mean at each time point is represented by a bold dash. (c and d) Phenotype of CD14+CD36+ monocyte population: (i) expression of Langerhans cell markers (CD1a and CD207) at day 1 (D1) and D4 post UVR (c); (ii) expression of dermal dendritic cell (DC; CD1c) and monocyte (CD206, CD209, CD163) markers at D4. Graphs show marker expression in a representative individual. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 UVR-induced activation and depletion of Langerhans cells (LCs). (a) Epidermal cell suspensions were analyzed by flow cytometry, and LCs were defined as viable CD45+ CD14− CD36− CD1ahigh CD207+ cells. (b) Time-course quantification of LCs in exposed versus unexposed epidermis. Results are expressed as the mean±SD ratio of the percentage of LCs in exposed (UV-LC) versus unexposed (C-LC) epidermal cells from the 27 volunteers. (c) CD86 and HLA-DR expression profile in gated LCs. Graphs show marker expression in a representative individual. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 Three distinct CD1a+ DC subsets populate UVR-exposed human epidermis. (a) Analysis of epidermal DC subsets according to their CD1a and CD207 expression profile allowed the identification of three DC subsets in the exposed skin: (i) CD1ahighCD207high (LC); (ii) CD1alow CD207− DC (IDEC); and (iii) CD1alow CD207+ DC (Pre-LC). (b) Time-course analysis of the percentage of the 3 epidermal DC subsets (IDECs, Pre-LCs, and LCs) following UVR exposure. Phenotypic identification was carried out by flow cytometry, as defined above. Data are expressed as percentage of cells in the viable 7AAD− CD45+ cell population from the 27 volunteers and the mean at each time point is represented by a bold dash. (c) DC subsets (IDECs, Pre-LCs, and LCs) infiltrating the exposed epidermis at D4 post UVR were analyzed for the expression of several surface markers (CD1c, CD209, CD206, and CD163). Graphs show marker expression in a representative individual. IDEC, inflammatory dendritic epidermal cell; LC, Langerhans cell; Pre-LC, inflammatory Langerhans epidermal cell. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 Correlations between the presence of monocytes, T lymphocytes, inflammatory dendritic epidermal cells (IDECs), and inflammatory Langerhans epidermal cells (Pre-LCs) in UVR-exposed epidermis at day 1 (D1) and day 4 (D4). Spearman’s rank cross-correlations between the frequencies of monocytes, T lymphocytes, IDECs, and Pre-LCs. Data are expressed as percentage of cells in the viable 7AAD− CD45+ cell populations extracted from the UV-exposed epidermis of each volunteer at D1 (a) and D4 (b) post UVR. Spearman’s correlation coefficient (rs) is shown in each figure. The solid line represents the regression line. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

7 Figure 6 CD1alow CD207− DC inflammatory dendritic epidermal cells (IDECs) and CD1alow CD207+ DC inflammatory Langerhans epidermal cells (Pre-LCs) show significant in vitro allostimulatory functions. Epidermal cells were collected from control (gray bars) and UV-exposed (black bars) skin samples of two healthy volunteers at D4, stained, and analyzed by flow cytometry. Cells were gated as viable CD1ahighCD207+ LCs, CD1alow CD207− IDECs, CD1alow CD207+ Pre-LCs, and CD1a−CD207−CD14+CD36+ monocytes and were purified by cytometry cell sorting and then added to allogenic T cells from two different normal human blood donors for 6 days. [H3]-thymidine was added during the last 18 hours of culture. Proliferation in control cultures (without APC subsets) is also shown (white bars). Results are expressed as count per minute (c.p.m.). The graphs show the Mean±SD obtained from triplicate cultures of both healthy volunteers EC cultured with allogeneic T cells from one donor and are representative of two independent experiments. APC, antigen-presenting cell; EC, epidermal cell. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

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