Figure 2.

YSA redirects adenovirus to the EphA2 receptor. A: Transduction of CAR-expressing A549 cells with wild-type, ablated and YSA-retargeted adenoviral vectors with 1000 gc/cell: wt-Ad-Luc, Ad-Luc/∆F(FG)∆P and Ad-Luc/∆F(FG)∆P-YSA. Twenty four hours after infection cells were lysed to determine luciferase levels. Data are expressed as mean ± SD (n = 3). B: Incubation with bi-specific antibody restores infectivity of ablated adenoviral vectors in human fibroblasts. Cells were transduced with 500 gc/cell of Ad-/∆F(FG)∆P or Ad-/∆F(FG)∆P-YSA with (black bars) or without (white bars) EGF receptor targeted bi-specific scFV molecules. Luciferase activity was measured after 24 h and data are expressed as mean ± SD (n = 3). C: Insertion of YSA peptide in the HI loop of ablated adenoviral vector partially restores transduction of human pancreatic cancer cell lines Capan-1 and MiaPaca-2. Cells were transduced with 1000 gc/cell of wt-Ad-Luc (white bars), or Ad-Luc/∆F(FG)∆P (gray bars) or Ad-Luc/∆F(FG)∆P-YSA (black bars). Luciferase activity was measured after 24 h. Data are expressed as mean ± SD (n = 3). D: Pre-incubation with synthetic peptide blocks YSA-mediated targeting of human pancreatic cancer cell line MiaPaca-2. Cells were preincubated with 250 (grey bars) or 500 (black bars) μmol/L synthetic YSA peptide and transduced with 500 gc/cell of Ad-Luc/∆F(FG)∆P or Ad-Luc/∆F(FG)∆P-YSA. Luciferase was measured after 24 h. Data are expressed as mean ± SD (n = 3).