FIGURE 2.

Differential APM component up-regulation in DC matured with different cytokine combinations. A, representative flow cytometry histograms for APM component expression are shown, as performed for 12-healthy donor DC preparations. Separate experiments were repeated for 2 or 3 times per donor. After 6 days of culture in interleukin-4 and granulocyte macrophage colony stimulating factor supplemented media, human monocyte-derived immature DC were treated with medium alone or medium cultured with 1 of the cytokine combinations listed in Table 1 for 48 hour at 37°C. DC were then harvested and stained intracellularly with mAb specific for δ, LMP2, TAP1, TAP2 or tapasin (as described in Materials and Methods section). Stained cells were analyzed by flow cytometry, adjusting the MFI for control isotype matched mAb-stained cells to 5, to permit comparison between different DC culture conditions. B, MFI values summarized from 12 sets of healthy, HLA-A*0201⁺ donor DC that were matured with different cytokine combinations listed in Table 1. Mean MFI values (± SEM) obtained by flow cytometry are shown, where the isotype control staining was set to MFI = 5, to permit comparison between different DC culture conditions (*P<0.05). APM indicates antigen processing machinery; DC, dendritic cell; mAb, monoclonal antibody; MFI, mean fluorescence intensity.