CD33/CD123 NANOBODY TCE mediates killing of AML blasts and expansion of T cells. Primary AML samples from relapsed patients (n = 18) were treated with indicated TCE for 7 days and specific apoptosis was calculated. (A) Violin plots displaying the percentage of SP in bulk (n = 18) and CD34+ AML cells (n = 14) and the fold change of CD3 cells to untreated control (n = 15). Lines in the violin plots indicating median and quartiles. (B) Treatment effects in 2 representative AML samples. Top row: baseline expression of CD33 and CD123 in histograms. Bar graphs displaying the ratio of mean florescent intensity (MFI) to isotype control and the baseline percentage positivity of CD33, CD123 and CD3; bottom row: bar graphs displaying SP apoptosis in bulk and CD34+ AML cells, and fold change of CD3+ T cells in day 7 samples. (C) Bar graph displaying the number of colonies formed in samples under different treatment, CD33/CD123-TCE at 25 nM, triplicate/treatment, mean ± standard deviation (SD). L: 2.5 nM and H: 25 nM. Plus signs (+) indicates the comparison between untargeted TCE and the indicated treatment with P < .001 (one-way analysis of variance [ANOVA] followed by Tukey test with adjust P values). CD3 fold change is not significant difference among the treatments.