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Fig. 8. Mature IL-4-DC and GM-IL-4-DC induce Th1 cytokines during primary allogeneic MLR. Mature IL-4-DCs and GM-IL-4-DCs (maturation was induced by TNF-
) were co-cultured with allogeneic NPBMC depleted of HLA-DR+ cells, NK cells and B cells as described in Materials and Methods at 1:10 ratio for 6 days, activated 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 washed, permeabilized and stained for detection of intracellular cytokines using FITC- or PE-conjugated specific antibodies. 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 four experiments with similar results is shown.
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