Figure 5

Induction of HLA-DPB1^*0501-restricted MAGE-A4-specific CD4⁺ T cells from donor 4 using a synthetic M41 peptide. (A) Donor 4-derived CD4⁺ T cells were primed with autologous DC pulsed with synthetic M41 peptide (10 μg ml⁻¹), but not rMAGE-A4 protein. The primed CD4⁺ T cells were stimulated with autologous PBMC pulsed with irrelevant peptide (open bar) or cognate peptide (filled bar) in the presence of 5 μg ml⁻¹ of mAbs against HLA-DP, HLA-DQ or HLA-DR. (B) The M41-specific Th cells were stimulated with allogeneic EBV-B-cell lines pulsed with irrelevant (open bar) or cognate (filled bar) peptide. All T-cell cultures were incubated with the MMC-treated target cells in duplicate for 20–24 h, then the culture supernatants were assessed for IFN-γ levels in ELISA. These experiments were carried out independently at least three times. Data denote mean±s.e.m. and are representative of the experiments.