FIGURE 1.

Characterization of the antigen reactivity of TIL 2035 T cells. A, TIL 2035 cells were co-cultured with either autologous melanoma line 2035mel (HLA-A2, A23) or HLA-mismatched control melanoma line 888mel (HLA-A1, A24) for 6 hours, followed by incubation with an anti-CD107a monoclonal antibody and FACS analysis. Stimulations carried out with 2035 mel are indicated by the bold lines, and those carried out with control 888 mel are indicated by the filled histograms. Gating was carried out on the cell populations that expressed the indicated BV gene products. B, The autologous melanoma line and HLA-A2-positive melanoma cell line, 624 mel, were incubated with either no antibody (Control), anti-HLA-A*02 antibody (Anti-A2) or anti-HLA-A23 antibody (Anti-A23). TIL 2035-F6/8 was then added, and 18 hours later IFNγ release was measured by ELISA. C, Autologous EBVB cells were pulsed with either a control peptide (upper left) or the HLA-A2-restricted NY-ESO-1: 157-165 peptide (upper right) for 2 hours, and then co-cultured with TIL 2035-F6/8 for 2 hours. An IFNγ capture assay was then carried out, cells were co-stained with an anti-BV17 antibody, and FACS analysis was performed. Two hundred ninety-three cells were co-transfected HLA-A23 with either a control vector (lower left) or a vector encoding MAGE-6 (lower right) for 24 hours, followed by the addition of TIL 2035-F6/8. Eighteen hours later, an IFNγ capture assay was carried out; cells were co-stained with an anti-BV1 antibody and analyzed using FACS analysis. The data represent one of three individual experiments.