Fig. 2

In vitro safety profile of TEG001. (a) Comparable recognition profile of Daudi tumor cells versus healthy hematopoietic cells. Effector cells TEG001 and target cells (E:T ratio 1:3) were incubated for 24 h in the presence of 10 μM PAM. IFNγ secretion was measured by ELISPOT. IFNγ spots per 15,000 T cells are shown as mean ± SD of at least 3 independent replicates for each target. Fifty spots/15,000 cells were considered as a positive recognition response and indicated by the black horizontal line. (b) RhoB distribution for healthy hematopoietic cells upon irradiation as analyzed by confocal microscopy in the presence of 10 μM PAM. Data is shown as fold-changed of RhoB distribution between irradiated cells (cellular stress condition) compared to non-irradiated cells from average ratio of at least ten different cells; (c) CD34⁺ progenitor cells from a healthy donor were treated with either 50 IU/ml IL-2, 1000 IU/ml IFNγ, 5 mM Cy, 20 μM Flu or combination of Cy/Flu. As positive control primary AML blast from donor p25 was treated with PAM. All cells were analyzed for intracellular distribution of RhoB using confocal microscopy. White bars represent healthy CD34⁺ progenitor cells, while black bar indicate primary AML blast (p25 AML). The RhoB signal ratio between nuclear and extranuclear cellular compartments was measured using ImageJ image analysis software. Graphs show average ratios of at least ten different cells ±SEM. Statistical significance compared to untreated CD34⁺ progenitor cells was determined by using Kruskal-Wallis test and Dunn’s multiple comparison test; (d) Comparable recognition profile of healthy hematopoietic cells in non-stressed (left panel) and stressed (irradiated, right panel) conditions. Effector cells TEG001 and target cells (E:T ratio 1:3) were incubated for 24 h in the presence of 100 μM PAM. IFNγ secretion was measured by ELISPOT. IFNγ spots per 15,000 T cells are shown as mean ± SD of at least 3 independent replicates for each target