Figure 3.

Functional characterization of the N^361-369-specific TCR 1 and TCR 4. (A) Flow cytometric analysis of the transfection efficiency of TCR 1 and TCR 4 in CD4⁺ and CD8⁺ T cells, by N^361-369-HLA-A∗11:01 tetramer. (B) Binding assay of TCR-transduced CD8^- and CD8⁺ Jurkat cells with N^361-369-HLA-A∗11:01 tetramer. (C) Flow cytometric analysis of T cell activation evaluated by CD137 expression. TCR-transduced CD8⁺ T cells and CD4⁺ T cells were individually co-cultured with HLA-A∗11⁺ K-562 cells pulsed with N^361-369, or an equimolar amount of DMSO, for 24 h. Expression of CD137 on CD8⁺ T cells (left), CD4⁺ T cells (middle) was evaluated and did further statistical analysis (right). (D) N^361-369 loaded HLA-A∗11:01⁺ K-562 cells lysis by TCR 1-CD8⁺ T cells after 8 h at different E:T ratios. DMSO was used as negative control. Reported values are the mean of triplicates, with error bars indicating. E, Effector T cells. T, Target cells. (E) Detection of IFN-γ, IL-2 and TNF-α secretion from TCR 1 and TCR 4-transduced CD4⁺ T cells by ELISA, after co-cultured with HLA-A∗11:01⁺ K-562 cells loaded with N^361-369 for 24 h. Data were presented as mean ± SD, n = 3, ∗∗P < 0.01, ∗∗∗P < 0.001. Representative data of two independent experiments were shown.