Figure 3

In vitro induction of CTL responses in cancer patient PBMCs. T cells were stimulated with autologous DCs treated as described under Methods. Briefly, stimulations were performed once every 8 days for a total of 4 weeks with cytokine additions- IL-2 and IL-7 (10 ng/ml each) and expanded after the third stimulation using anti-CD3/anti-CD28 beads. Expanded cells were harvested on day 14 and restimulated with DC+ vaccine (B11-hCGβ, 10 μg/ml) followed by activation with different stimuli (3a-c donor #s 1–3) or R-848 (0.1 μM or 0.3 μg/ml.) and Poly IC (50 μg/ml) added alone or in combination (3d, donor #s 4–6). T cells were assayed for effector function on days 5 and 6 post stimulation using the Granzyme B ELISpot assay kit.