Figure 2

Induction of HER-3-reactive CD4 T cells.

(A) HER-3-reactive CD4 T cells were elicited in 4 healthy individuals (H16 is from donor1: DR4/9, DR53; T11 is from donor2: DR9/12, DR53; O3 is from donor3: DR9/13, DR53; n24 is from donor4: DR9/13, DR53) and then tested for their ability to produce IFN-γ in response to HER-3 peptides. Peptides (0–30 μg/ml) were loaded on irradiated autologous PBMCs. Points: means of triplicate measurements, bars: SEM. The results shown are representative of 3 experiments that were performed on the same samples. (B) We used an IFN-γ assay to assess the responses of HER-3-reactive CD4 T cells to HER-3 peptide (3 μg/ml)-loaded irradiated autologous PBMCs. L243 (10 μg/ml, an anti-HLA-DR antibody) or W6/32 (10 μg/ml, an anti-HLA Class I antibody) was added to confirm the HLA-DR restriction of the T-cell responses. (C) L-DR4, L-DR9 or L-DR53 cells with or without HER-3 peptide (3 μg/ml) were used as antigen-presenting cells (APCs) to evaluate the specific HLA-DR restriction of each HER-3-reactive CD4 lines by means of an IFN-γ assay. Columns: means of triplicate measurements, bars: SEM. The results shown are representative of 3 separate experiments.