Identification of blood DCs in cultured PBMCs.

DCs were defined as cells that were negative for CD3, CD14, CD16, CD19, and CD34 (Lin⁻), and express HLA-DR (HLA-DR⁺); they represented a distinct cell population in cultured PBMCs for up to 3 days (right lower quadrants in A). The expression of CD40, CD80, CD86 costimulatory molecules (B); CMRF-44, CMRF-56, and CD83 activation markers (C); and CD11c⁺ and CD123⁺subset markers (D) on gated Lin⁻ HLA-DR⁺ DCs was assessed by using 3-color flow cytometry. Isotype control mAb staining is shown (broken line).