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. 2021 Oct 27;13(21):5390. doi: 10.3390/cancers13215390

Figure 3.

Figure 3

The HLA-A11-restricted dNPM1 TCR targets primary AML. CD8 T-cells transduced with the dNPM1 or EBV TCR were generated from two HLA-A11+ healthy individuals (donors 10095 and 10231). Results are shown for donor 10095. Results for donor 10231 are shown in Figure S4. (A) T-cells were incubated overnight with five HLA-A11+ primary AMLs at different E:S ratios and IFN-γ secretion was measured by ELISA. AML cells were also pulsed with a mix of dNPM1 and EBV peptides at a concentration of 500 nM per peptide. T-cells with the dNPM1 TCR (red bars) and clone 6F11 (green bars) reacted against dNPM1 AMLs, while wtNPM1 AMLs were not recognized. T-cells with the EBV TCR (blue bars) recognized all five AMLs after peptide loading. Bars represent mean + SD of duplicate wells; (B) A 9-h chromium-51 release assay was performed to test T-cell cytotoxicity against five HLA-A11+ primary AMLs at different effector:target (E:T) ratios. T-cells with the dNPM1 TCR (red line) and clone 6F11 (green line) lysed dNPM1 AMLs, while wtNPM1 AMLs were not killed. T-cells with the EBV TCR (blue line) did not show lysis of primary AMLs. Symbols represent mean ± SD of triplicate wells.