Fig. 9. Immature IL-4-DC and GM-IL-4-DC induce IL-10 but suppress IL-2 and IFN- from allogeneic bulk T cells after repeated stimulations. IL-4-DCs and GM-IL-4-DCs were co-cultured with allogeneic NPBMC depleted of HLA-DR⁺ cells, NK cells and B cells for 6 days at a ratio of 1:10. Stimulation with APCs was repeated weekly from the same donor under the same conditions for three times as described in Materials and Methods. Six days after the last stimulation with APCs, cells were washed, re-stimulated with 2.4 µg/ml PHA plus 1.0 ng/ml PMA for 6 hours. Brefeldin A was added for the last 4 hours. Cells were permeabilized, stained for intracellular cytokines using FITC- or PE-conjugated specific mAbs. Lymphocytes were gated and intracellular cytokines were analysed by flow cytometry. Numbers within histograms represent percentages within marker bounds. Markers were set employing isotype control staining. One of three experiments is shown.