Induction of the DC-associated differentiation/activation antigens CMRF-44, CMRF-56, and CD83 on cultured Lin⁻ cells.

Sort-purified CD11c⁺ and CD11c⁻ populations were cultured for 18 hours in the presence of GM-CSF and IL-3 with or without allogeneic T lymphocytes. Following culture, cells were harvested and stained with biotinylated CMRF-44 or CMRF-56, or purified CD83 mAb followed by biotinylated anti–mouse IgG. Biotinylated antibody was detected with PE- or PE-Cy5–conjugated streptavidin, in conjunction with PE-Cy5– or PE-conjugated CD16 or CD34 for the (A) CD11c⁺ and (B) CD11c⁻ populations, respectively. CD11c⁺CD16⁻, CD11c⁺CD16⁺, CD11c⁻CD34⁺, and CD11c⁻CD34⁻ populations were gated and examined for their expression of activation antigens (solid lines). Dashed lines represent negative control antibody staining.