DC-10 are present in peripheral blood and can be differentiated in vitro from peripheral monocytes. (A) Expression levels of CD11b, CD11c, CD14, CD16, CD40, CD68, CD80, CD83, CD86, HLA-DR, CD115, CD163, and M-DC8 in gated CD14⁺CD16⁻ cells (inflammatory monocytes), CD14^lowCD16⁺ cells (resident monocytes), CD14^brightCD16⁺ cells (containing DC-10), myDCs (CD1c⁺CD11c⁺), and pDCs (CD11c⁻CD303⁺) in peripheral blood were evaluated by FACS analysis. Black histograms represent staining with the isotype-matched control mAbs. A representative donor of 10 independent donors analyzed is presented. Percentages of CD14⁺CD16⁻, CD14^lowCD16⁺, CD14^brightCD16⁺, CD1c⁺CD11c⁺, CD303⁺CD11c⁻ cells in total peripheral blood mononuclear cells, and percentages of CD14⁺CD16⁻, CD14^lowCD16⁺, CD14^brightCD16⁺, CD1c⁺CD11c⁺, CD303⁺CD11c⁻ cells expressing the indicated markers are indicated. (B) Morphology (May-Grünwald-Giemsa staining) of CD14⁺CD16⁻ cells (inflammatory monocytes), CD14^lowCD16⁺ cells (resident monocytes), CD14^brightCD16⁺ cells (containing DC-10) FACS-sorted from peripheral blood. Results from one representative donor of 2 tested is shown. (C) Monocyte-derived DCs were differentiated in IL-4 and GM-CSF in the presence of IL-10 for 7 days (DC-10), or in IL-4 and GM-CSF for 7 days (iDC), or in IL-4 and GM-CSF for 5 days and activated for additional 2 days with LPS (mDC). Morphology of DCs was evaluated by light microscopy and by May-Grünwald-Giemsa staining. Results from one representative donor of 5 tested is shown. Image information: Nikon Eclipse TE200 microscope; magnification 40×/0.55; cover slides were mounted using DXP (water free mounting medium); Nikon DXM1200 camera; Nikon ACT-1 Version 2.63 and PowerPoint software. (D) Expression of CD14, CD16, HLA-DR, CD40, CD83, CD80, CD86, CD163, CD1a, CD115, CD68, and M-DC8 was evaluated by FACS analysis. A representative donor of at least 15 tested is presented.