Figure 1

DC differentiation from monocytes induced by column activation. CTCL cells and DC were isolated from a leukapheresis by CD4 or CD3-antibody conjugated to magnetic beads. The cells were separated by passage through a column placed in a magnetic field and the purified CTCL cells were re-added to the column activated monocytes and cultured overnight. Binding of fluorochromes was analyzed using flow cytometry and 2-color quadstats were gated on the monocyte population. The results demonstrate membrane CD14-FITC and CD86-PE co-expression on cells obtained a: Day 0, primary isolation; and after overnight culture of b: leukapheresis cells; c: cells obtained by CD4-magnetic bead isolation and re-cultured overnight with column activated monocytes; and d: cells obtained from CD3-magnetic bead isolation and re-cultured overnight with column activated monocytes. e: Bar graph showing the reduction in mean fluorescent intensity (MFI) of CD14 expression on primary isolation (Day 0) and after overnight incubation of the leukapheresis (leuk) or column passaged and recombined cell populations using CD4 or CD3-magnetic bead isolation (negative control isotype staining is presented in the first bar). f: Bar graph showing the increase in MFI of CD86 expression (as described in e).