4.

DC quality control. Monocytes from healthy individuals were enriched from leucocyte apheresis products by different enrichment procedures, as indicated, differentiated into DCs followed by activation with LPS/IFN-γ. DC preparations using AIM-V/Octaplas or CellGro DC medium are combined. (A) The left-hand panel shows the ratio of increase in the expression density ± SEM of the indicated DC maturation markers measured 6 hrs (smDCs) and 48 hrs (mDC) after activation. DCs generated from elutriated monocytes (number of preparations, n= 35) are analysed. The right-hand panel compares the expression density mean fluorescence intensity (MFI) ± SEM of maturation markers measured on mDCs that are generated from monocytes isolated by the indicated enrichment procedures. MFIs of the isotype controls are below 5 (data not shown). (B) Secretion of IL-12 ± SEM secreted from mDCs analysed in (A) is illustrated. (C) Proliferation of allogeneic PBMCs in co-cultures with the DCs analysed in (A) is given relative to PBMCs stimulated with the super-antigen staphylococcal enterotoxin A/B (SEA/SEB) (normalized to 100%) and the background proliferation of unstimulated PBMCs (normalized to 0%). Number of preparations in (B) and (C): adherence, n= 10; selection, n= 8; depletion, n= 11, elutriation, n= 35. Comparisons of the enrichment procedures in (A) to (C) show no statistically significant differences.