AURKA_207-215–specific TCR-transduced CD8⁺ T cells can distinguish leukemia cells from normal cells on the basis of AURKA expression levels. (A) AURKA_207-215–specificTCR-transduced CD8⁺ T cells exhibit antileukemia reactivity in an HLA-A*0201-dependent fashion. The HLA-A*0201⁺ leukemia cell line GANMO-1 was lysed by AURKA_207-215–specificTCR-transduced CD8⁺ T cells as a function of effector:target (E/T) ratio; no significant lysis was observed with the HLA-A*0201⁻ leukemia cells lines MEG01, KAZZ and OUN-1. All of the tested leukemia cell lines overexpress AURKA mRNA; numbers in parentheses indicate AURKA mRNA expression relative to K562, and correlations with AURKA protein expression are shown in supplemental Figure 2. (B) The same AURKA_207-215–specificTCR-transduced CD8⁺ T cells used in panel A at the same E/T ratios were tested in ⁵¹Cr-release assays for potentially damaging effects against normal cells. No significant lysis was observed with HLA-A*0201⁺ PBMCs (n = 3), PHA-lymphoblasts representing normal mitotic cells (n = 3) or normal cord blood–derived CD34⁺ cells (CB-CD34⁺) encompassing normal hematopoietic progenitor cells (n = 2). AURKA mRNA expression relative to K562 was 0.02 ± 0.008 for PBMCs, 0.25 ± 0.005 for PHA-lymphoblasts and 0.21 ± 0.09 for CB-CD34⁺ cells (* indicates less than detectable). (C) Effects of HLA class I and class II blockade on the cytotoxic activity of AURKA_207-215–specificTCR-transduced CD8⁺ T cells against GANMO-1 leukemia cells. E/T, effector:target ratio. (D) As for panel C, showing the effects of HLA class I and class II blockade on the lysis of autologous B-LCLs loaded with AURKA_207-215 peptide (1μM). (E) Flow cytometric confirmation of HLA-A*0201 expression by MEG01-A2 cells. (F) Enhanced lysis of MEG01-A2 cells relative to parental MEG01 cells by AURKA_207-215–specificTCR-transduced CD8⁺ T cells confirms recognition of endogenously processed AURKA_207-215 peptide presented in the context of HLA-A*0201. E/T indicates effector:target ratio. Error bars represent SDs.