Fig. 5.
Restored production of IL-12 in monocytes obtained from PBMCs mobilized with G-CSF by culturing with IL-4 and GM-CSF. To assess the ability to produce Th-1-type cytokines from DCs generated from monocytes obtained after G-CSF mobilization, DCs were stimulated with 10 μg/ml LPS. DCs were purified with the use of Human Dendritic Cell Enrichment Cocktail (Stem Cell Technologies), which is a depletion cocktail containing several mAbs (anti-CD3, anti-CD14, anti-CD16, anti-CD19, anti-CD56, anti-CD66b, and Glycophorin A), according to the manufacturer’s instruction. Cells were then cultured at the concentration of 1 × 105/ml for 48 h. The concentrations of IL-10 and IL-12 were measured by ELISA using the harvested cell-free supernatants. Data represent the means ± SD, derived from two independent experiments using PBMCs obtained from patient nos. 1 to 3 and 5. There were no significant differences with regard to the production levels of IL-10 and IL-12 between PBMCs obtained before and after G-CSF mobilization