WT1-TCB* vs WT1-specific T cells. (A) Specific lysis of unpulsed and RMF peptide–pulsed primary AML cells from HLA-A*02⁻ and HLA-A*02⁺ patients by WT1_RMF-specific T-cell clone after 3 days in feeder layer–based long-term cocultures. Specific lysis was calculated by subtraction of unspecific lysis by HD T cells under the same conditions (n = 6-9). (B) Specific lysis of primary AML cells after 3 days in feeder layer–based long-term cocultures with WT1_RMF-specific T-cell clone and treatment with WT1-TCB* compared with HD T cells (n = 9). (C) Specific lysis of primary AML cells (all NPM1 wild-type) after 3 days in feeder layer–based long-term cocultures with WT1_RMF-specific T-cell clone and treatment with WT1-TCB*. Specific lysis was calculated by subtraction of unspecific lysis by a T-cell clone targeting an unrelated epitope of mutated NPM1 (n = 6 ). (D) Representative dot plots from flow cytometry analysis showing CD2 and CD33 expression in lysis experiments. Bars represent mean ± SEM. Statistical analysis: Mann-Whitney U test. *P < .05, **P < .01. n.s, not significant.