Functional analyses of the various HA-2 TCR-modified T-cell clones. (A) HA-2 TCR-modified T-cell clones were tested in a cytotoxicity assay against HLA-A2-positive HA-2-positive target cells (EBV-RZ; ▪) and HLA-A2-positive HA-2-negative target cells (EBV-Z; □) at an E/T ratio of 10:1. The cytotoxicity assay was performed for 4 hours. (B) HA-2/HLA-A2 tetramer staining of the HA-2 TCR-modified T-cell clones, shown as MFI, correlated with the lytic activity against HLA-A2-positive HA-2-positive target cells (EBV-RZ). (C) HA-2/HLA-A2 tetramer staining and CD8 expression of 5 representative HA-2 TCR-transferred T-cell clones^1-5  and one GFP/NGFR transduced T-cell clone is shown. (D) IFN-γ production of the HA-2 TCR-modified T-cell clones upon stimulation with HLA-A2-positive HA-2-positive stimulator cells (EBV-RZ; ▪) and HLA-A2-positive HA-2-negative stimulator cells (EBV-Z; □) was measured after 24 hours of stimulation. The original HA-2-specific T-cell clone HA2.27 was used as positive control in these experiments. The HA-2/HLA-A2 tetramer staining of HA2.27 was also determined (MFI = 474).