Figure 4.

NY-ESO-1 regions recognized by CD4 and CD8 T cells in patient GO. MACS beads-purified CD4 and CD8 T cells (2 x 10⁶) from PBMCs obtained in August 2005 were cultured with irradiated (40 Gy) autologous CD4- and CD8-depleted PBMCs (2 x 10⁶) as APCs in the presence of 28 overlapping 18-mer peptides and a 30-mer C-terminal peptide (OLPs) at a concentration of 1 µg/ml of each peptide in a 24-well culture plate for 14 days. On the twenty-sixth day after two stimulations, CD4 and CD8 T cells (3 x 10⁴) were assayed for IFNγ secretion against PFA-treated autologous CD4- and CD8-depleted PBMCs (3 x 10⁴) pre-pulsed with the individual peptide after stimulation for 4 h. The peptide number corresponds to the individual overlapping peptides. Abbreviations: P, positive control stimulated with a mixture of OLPs; N, negative control without peptides.