Schematic of acDC-based T-cell stimulation. Either unfractionated PBMCs (fresh or frozen) or undiluted heparinized whole blood were incubated with GM-CSF and IL-4 for 48 hours in the presence of protein Ags. Maturation stimuli were then added during the last 24 hours, after which amplified T-cell responses were measured by a variety of readouts. Top panels list T-cell readouts with PBMCs; bottom panels show readouts with whole blood. The acDC amplification technique is also compatible with peptide Ags, which were added at 24 hours along with maturation stimuli.