Comparison of mature MDDC derived from cryopreserved PBMC of healthy donors vs. cancer patients. A. Surface phenotype: Expression levels (MFI) of CD86, CD83, and HLA-DR on mature MDDC derived from healthy donors (healthy) were compared to those derived from cancer patients (cancer). B. Function: Mature MDDC from each group were cultured for additional 18–20 h in presence of BFA. Cells were processed and analyzed to evaluate the expression of intracellular IL-12 (%CD209+IL-12+) or COX-2 (%CD209+COX-2+) as described earlier. Quantities of secreted IL-8 (pg/ml) by mature MDDC from each of these two groups were detected by CBA assay of the culture supernatants collected on day 7. C. T cell stimulation: The top scatter plot shows proliferation of allogeneic CD4+ T cells using mature MDDC from PBMC of healthy donors and cancer patients. The lower two scatter plots demonstrate enhancement of MDDC mediated SEB-specific autologous CD4+ and CD8+ T cell proliferation. Both allogeneic and autologous antigen-specific T cell stimulation assays were set up and percent proliferation was measured as described earlier. Bars in all the scatter plots represent medians. *, statistically significant differences (P < 0.05); **, statistically significant differences (P < 0.01).