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The keratinocytes were identified by double immunofluorescence with IL-20 and CK14 (Fig.?1a). In some cases, strongly IL-20-positive cells on the dermal CAPNS1 side of the basement membrane were located in very close proximity to the CK14-positive keratinocytes in basal epidermis (insert in Fig.?1a). To identify the phenotype of the IL-20+ cells, we performed double staining for IL-20 and selected markers for T cells, DCs, endothelial cells and NK cells. Virtually, all of the IL-20+ cells in the papillae were also CD4+ (Fig.?1d), which is present on T helper and regulatory cells, DCs, monocytes, NK cells and macrophages [15]. The IL-20+ cells were negative for the T-cell markers CD3 (Fig.?1f) and CD8 (not shown) as well as for the NK cell marker CD56 (Fig.?1g), so we excluded the IL-20+ cells from being T cells or NK cells None of the IL-20+ cells were CD31+ and were thus not endothelial cells (Fig.?1e). The IL-20+ cells did not express HLA-DR (Figure?S2a), and we concluded that they were not antigen-presenting cells. Except for a few double-positive cells, the main part (>95%) of the IL-20+ cells were CD11c? (Fig.?1h) and CD68? (Figure?S2b), which means that the IL-20+ cells were not myeloid DC nor macrophages. Interestingly, more than 50% of the IL-20+ cells in the dermal papillae were positive for CD1a (Fig.?1i and Figure?S2b,c). Furthermore, the dermal IL-20+/CD1a+ cells were clearly different in morphology from the epidermal IL-20?/CD1a+ cells. The latter were larger and carrying learn more a Talazoparib supplier more intense CD1a staining (Fig.?1i). Double stainings for IL-20/langerin revealed that approximately 90% of the IL-20+ cells were langerin+. As for CD1a, the morphology of the dermal IL-20+/langerin+ cells was clearly different from the morphology of the epidermal IL-20?/langerin+ cells. The intensity of langerin staining was much lower in the dermal IL-20+/langerin+ cells compared with the epidermal IL-20?/langerin+ Langerhans cells. Further characterization of the IL-20+ cells showed that only a minor part (5�C10%) were also positive for the activation marker CD83 (Fig.?2a), and