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Figure 5 | Journal of Experimental & Clinical Cancer Research

Figure 5

From: Characterization of an engineered human purine nucleoside phosphorylase fused to an anti-her2/neu single chain Fv for use in ADEPT

Figure 5

hDM-αH-C6.5 MH3B1 specifically associates with HER2/ neu expressing cells and causes cytotoxicty in the presence of F-dAdo irrespective of expression of tumor antigen or cell growth rate. (A), hDM-αH-C6.5 MH3B1 associates with HER2/neu expressing cells resulting in concentration dependent cytotoxicity upon addition of 1.5 or 6 μM F-dAdo to CT26HER2/neu or MCF-7HER2 cells respectively. Different concentrations of hDM-αH-C6.5 MH3B1 were incubated with cells, unbound enzyme washed away, F-dAdo added and 72 hours later cellular proliferation was determined by MTS assay. (B), CT26HER2/neu and CT26 cells were seeded at different ratios and grown overnight. hDM-αH-C6.5 MH3B1 was incubated with cells for 45 minutes, and washed away. Cells were then grown in the presence of 1.5 μM F-dAdo for 72 hours and cell proliferation determined by MTS assay. (C), MCF-7HER2 cells were grown overnight (O/N) in the presence of 10% serum, washed and growth continued for 72 hours in the presence of varying amounts of serum. The column labeled overnight (O/N) represents the number of cells prior to switching to different amounts of serum. The number of cells was determined by visually counting the cells. (D), Viability of MCF-7HER2 cells in the presence of different amounts of fetal bovine serum and 1.5 μM F-Ade was determined after 72 hours of incubation by MTS assay. Error bars for each graph represent standard deviation within each set of values.

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