Sensitivity of Dendritic Cells to Microenvironment Signals

Figure 3

Leukemic cell-derived proteins effect on monocytes differentiating into dendritic cells. Mononuclear cells were obtained after gradient density from buffy coats of healthy individuals. After 2 h of adhesion, lymphocytes were removed and monocyte cultures were stimulated with IL-4 and GM-CSF (50 ng/mL) to induce dendritic cell differentiation. K562 supernatants (SN K562) were obtained after 3 days of cell culture in RPMI plus 10% of fetal bovine serum (FBS) followed by filtering (0.22 μm). Part of tumor supernatant and part of IL-4 and GM-CSF were boiled for 10 minutes at 100°C in order to denature proteins. After cooling, some cells were incubated with these supernatants or cytokines at the same concentration described above for 5 days. Anti-CD14 FITC and anti-CD1a PE were used to stain the cells and data were assessed by a FACS Calibur. Graphs show mean ± SEM of the percentage of CD14⁺ (a) and CD1a⁺ (b) cells. ; ; . .

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