Cell-surface phenotypes of CD4⁺/HLA-DR^hi and CD4⁺/HLA-DR^lo human dendritic cell precursors before and after in vitro culture in the presence of human GM-CSF and TNF-α.

(A) BM cells obtained from NOD/SCID mice that had been inoculated with human CBMNCs were stained with fluorescein isothiocyanate (FITC)-labeled irrelevant murine IgG₁, anti-CD3, anti-CD11a, anti-CD14, anti-CD34, or anti–HLA-DQ antibody, and phycoerythrin (PE)-labeled anti–HLA-DR antibody and biotinylated anti-CD4 antibody plus streptavidin-RED 670, or, for analysis of CD11c or CD83 expression, with PE-labeled irrelevant murine IgG₁, anti-CD11c, or anti-CD83 antibody, and FITC-labeled anti–HLA-DR antibody and biotinylated anti-CD4 antibody plus streptavidin-RED 670. Data acquisition and analysis were performed with Cellquest software (Becton Dickinson). The solid and dotted lines show specific antigen expression and the isotype-matched controls, respectively. The positive rates (mean ± standard error) obtained from 3 independent experiments are represented in the panels. (B) Cells in the CD4⁺/HLA-DR^hi fraction and the CD4⁺/HLA-DR^lo fraction were sorted and cultured for 7 days in the presence of 100 ng/mL GM-CSF and 10 ng/mL TNF-α, and the cultured cells were stained with FITC-labeled CD14 antibody and PE-labeled CD1a antibody. A replicate experiment yielded similar results.