GLEA2-specific CTLs were tested for their ability to lyse U251 cells expressing GLEA2. a GLEA2-specific cytotoxic T lymphocytes (CTLs) were generated in vitro with 5 μg/ml of anti-CD3 antibody (clone 17A2; Biolegend) pre-bound to the culture plate and 2 μg/ml of soluble anti-CD28 antibody (clone 37.51; Biolegend) at 37 °C for 3ds. GLEA2-specific CTLs were tested against U251 cells. b GLEA2-specific cytotoxic T lymphocytes (CTLs) were generated by weekly stimulation of nonadherent peripheral blood lymphocytes with irradiated, autologous DCs transduced with Ad-GLEA2. GLEA2-specific CTLs were tested against U251 cells lacking GLEA2 expression. CTLs generated from Ad-LacZ transduced DCs and effector cells generated from T lymphocytes stimulated by PBS were used as controls. Experiments performed in triplicate showed consistent results.