Phenotypical and functional characteristics of IFN-DC in comparison to IL-4/TNF-DC. (a) Morphology of IFN-DC (left) and IL-4/TNF-DC (right). Cells were stained according to Pappenheim and viewed at x630 magnification. (b) Immunophenotypic analysis of IFN-DC and IL-4/TNF-DC. DC were gated according to their FSC/SSC characteristics. The results are summarized as mean ± SD of % PE positive CD45+ cells. (c) Stimulatory potential of DC in an allogeneic MLR. Different numbers of DC or monocytes were cocultured with 1 × 105 PBMC for 4 days in the presence of anti CD28 and CD49d antibodies. 3 [H]-Thymidin was added, and incorporated radioactivity was measured by β-scintillation. Results are expressed as the mean ± SD of the stimulation index. (d) Cytokine production of allogeneic T cells. Allogeneic PBMC were cocultured in a 2:1 ratio with monocytes, IFN-DC or IL-4/TNF-DC or without any stimulus for three days. IFN-γ (left) and IL-4 (right) expression was detected by intracellular flow cytometry. The results are summarized as mean ± SD of % cytokine positive CD3+ cells after subtraction of background levels. (e) Migratory capacity of DC. Migration was measured in 24-well transwell culture chambers. Each well was loaded with 2.5 × 105 cells. The proportion of migrated cells towards Mip-3β as measured by flow cytometry is summarized as mean ± SD. Significant differences (p < 0.05) are marked by an asterisk. The brackets and asteriscs in c refer to the highest effector cell number.