Identification of Two Novel HLA-A∗0201-Restricted CTL Epitopes Derived from MAGE-A4
Figure 2
Induction of peptide-specific CTLs from human PBMCs. PBMCs from healthy HLA-A*0201 donors were first stimulated with peptide-pulsed autologous DCs and then restimulated with peptide-pulsed autologous PBMCs. After three to five days of the final stimulation, the stimulated PBMCs were used as effector cells to detect their cytotoxic activity against tumor cells at the indicated E/T ratios in a cytotoxicity assay. The irrelevant peptide HBcAg18–27 was taken as negative control. Each sample was measured in three replicates and the experiment was repeated three times. (a) P1, P2, P3, and P4-stimulated PBMCs mediated lysis of LB1751-MEL cells. (b) P1- and P3-stimulated PBMCs mediated lysis of LB1751-MEL cells (P1 and P3) and LB1751-MEL cells which surface HLA-A*0201 molecules were blocked with ant- HLA-A*0201 mAb (P1+HLA-A2 mAb and P3+HLA-A2 mAb). (c) P1- and P3-stimulated PBMCs mediated lysis of MCF-7A4 cells. (d) P1- and P3-stimulated PBMCs mediated lysis of MCF-7 cells.
