Figure 3. Correlation between TCR functional avidity and effector functions in the TCR-transduced CD8⁺ T cells.

(A) Peptide concentration-response curves in 2D3 cells transduced with four different mWT1₂₃₅-specific TCRs (B10-, TM-H2-, FSK1-, and TM-L1-TCRs) for the stimulation with titrated concentration of mWT1₂₃₅. Y-axis represents the frequency (% of max) of GFP-positive cells in 2D3 cells. (B) Cytokine production of mWT1₂₃₅-specific TCR-transduced CD8⁺ T cells stimulated with 1 μg/ml of mWT1₂₃₅. Frequencies of IFN-γ-single-, TNF-α-single-, and IFN-γ and TNF-α-double-producing CD8⁺ T cells are shown. (C) Proliferation of WT1-specific TCR-transduced CD8⁺ T cells. CD8⁺ T cells (5 × 10⁵ cells) were weekly stimulated with mWT1₂₃₅, and the estimated number of the WT1-specific TCR-transduced CD8⁺ T cells was calculated every week. All data are mean values ± SEM (n = 3). (D) Peptide concentration-response curves in 2D3 cells transduced with mWT1₂₃₅-specific B10- or TM-H2-TCR for the stimulation with titrated concentration of nWT1_235. It should be noted that peptide used for making the peptide concentration-response curve was mWT1₂₃₅ for (A) and nWT1₂₃₅ for (D). (E) Cytotoxic activity of B10-TCR or TM-H2-TCR-transduced CD8⁺ T cells against WT1-expressing leukemic cells. Assay for cytotoxic activity was performed repeatedly and the representative results are shown. All data are replicate measurements and represent mean values ± SEM.