Purification of DC1 and DC2.

Mononuclear cells were enriched for DC by the depletion of CD14⁺ monocytes and CD19⁺ B cells, followed by the positive selection of HLA-DR-expressing cells using immunomagnetic beads, as described in “Materials and methods.” HLA-DR⁺ cells were labeled with anti-CD11c PE, anti-IL-3Rα biotin plus streptavidin TC, and FITC-conjugated antibodies to lineage markers CD3, CD14, CD16, CD19, CD20, CD34, CD56, and IgM. CD11c⁺, FITC⁻ DC1 (left) and IL-3Rα^bright, FITC⁻ DC2 (right) were sorted on a FACSVantage using the gates shown in (A). DC1 and DC2 were reanalyzed after separation for purity assessment (B). Results are representative of more than 10 experiments.