Figure 9.
Functional activation of lymphocytes migrated to alternatively matured DC. a, b 5 × 105 mature DC were seeded in 12 well plate for 4 h, after which 3 × 106 autologous PBMC were added above a 3 μm insert. After 18 h incubation the cells that had migrated into the bottom chamber were taken and either immediately evaluated for IFNγ secretion (a) or incubated for 4 h in the presence of Daudi cells and evaluated for CD107a degranulation (b). c 5 × 105 mature DC were pulsed with SEB for 30 min at 37°C, washed and then seeded in 12 well plate for 4 h, after which 3 × 106 autologous PBMC were added above a 3 μm trans-well insert. After 4 h incubation the cells in the bottom chamber were taken and evaluated for IFNγ secretion. Shown is one representative experiment out of two (c) or three (a, b) with similar results.