Isolation of DC-SIGN⁺ blood cells from PBMCs.

(A) Total blood was stained with anti-CD14–PE and anti–DC-SIGN–FITC antibodies or with IgG1 isotype control antibodies coupled to FITC. (B) PBMCs were immunomagnetically depleted of T, B, and NK cells, using anti-CD3, anti-CD20, and anti-CD56, respectively, followed by positive flow cytometric sorting of DC-SIGN⁺ cells stained with FITC-conjugated anti–DC-SIGN (AZN-D1) antibody. The number of cells at each isolation step is shown. (C) DC-SIGN⁺ blood cells were obtained with more than 90% purity, as shown by re-examination of the cell population isolated as described in panel B. A representative experiment is shown.