CTL clone, PHYI8/63, generated by DCs treated with CHP/HER2-147, is specific for HER2p63.
(A) To assay the cytotoxicity of PHYI8/63 cells, 1 × 104HER2-expressing ovarian cancer cells, SKOV3, transfected with HLA-A2402 cDNA (SKOV3-A24), SKOV3 transfected with HLA-A0201 cDNA (SKOV3-A2), and HLA-A2402− HLA-A0201− parental SKOV3 were incubated with PHYI8/63 cells at serial effector-target ratios for 5 hours. Cytotoxic activity was measured using 51Cr release assay. (B) HLA-A2402+HER2− T2-A24 cells (1 × 104) were pulsed with 10 μM HER2p63 (HER2p63/T2-A24) or HER2p780 (HER2p780/T2-A24) for 1 hour at room temperature and for an additional hour at 37°C. PHYI8/63 cells were incubated with HER2p63/T2-A24, HER2p780/T2-A24, or T2-A24 at serial effector-target ratios for 5 hours. Cytotoxic activity was measured using 51Cr release assay.
