FIG. 1.
Preparation of monocyte-derived dendritic cells (DC). (A) Plastic-adherent PBMC (>80% CD14+) was differentiated in DC medium for 5–7 days. (B) Greater than 95% of high forward versus side scatter cells in the DC differentiation culture expressed high levels of CD11c and MHCII. DC derived from adherent monocytes (see Materials and Methods section) were defined as large cells (relative high forward and side scatter profiles by flow cytometry) expressing high levels of surface CD11c and MHCII. (C) Cells expressing high surface CD11c readily phagocytosed soluble ovalbumin (OVA) antigen. Fluorescence microscopy showing cells with high surface expression of CD11c (arrow), and “OVA” arrow indicates APC-conjugated OVA that has been endocytosed by CD11chi cells. Similar results were obtained in >10 additional experiments. PBMC, peripheral blood mononuclear cell.