Efficient SAD in haploidentical and single HLA allele mismatch settings. (Left) CD8 T cells of healthy donor 308 were stimulated with haploidentical MZ431-AML blasts. (Right) CD8 T cells of HLA-A*0302^pos healthy donor 410 were stimulated with BA85-RCC cells expressing A*0301 as the only HLA class I mismatch allele (for complete HLA types see Table 1). (A) IFN-γ ELISPOT results of day 18 MLLC/MLTC responders tested against AML/RCC cells and against K562 transfectants expressing the recipients HLA mismatch alleles. (B) Following 24 hours of incubation with K562-A*0201 or K562-A*0301, day 22 MLLC/MLTC responders were depleted of mismatch-reactive CD8 T cells using the CD137/CD65 approach. The resulting CD137/CD65^neg fractions (white bars), the CD137/CD65^pos fractions (gray bars), and the MLLC/MLTC controls stimulated exclusively with AML/RCC cells (black bars) were tested in IFN-γ spot assay. (C) The CD137/CD65^neg fractions (white bars) and the MLLC/MLTC controls (black bars) were reevaluated on day 33 after stimulations with leukemia or tumor cells on day 22 and day 29.