FIG. 5.

Ad-ISF35 IDI-mediated induction of late T-cell infiltration and humoral–cellular immune response. (A) Analysis of tumor-infiltrating lymphocytes and (B) their expression of CD25. (C–F) Serum and splenocytes from mice cured by Ad-ISF35 treatment were tested in vitro. (C) Sera from Ad-ISF35-treated mice, but not control animals, contained antibodies that recognize A20-derived tumor proteins detected by Western blot. A single-well electrophoresis was performed with A20 protein lysate. (D) Complement-dependent cytotoxicity (CDC) of A20 cells was determined using sera from A20-bearing mice treated with the vehicle, Ad5, or Ad-ISF35. As a control, serum was inactivated at 56°C for 1 hr. (E) A20 cell apoptosis induced by splenocytes from vehicle-, Ad5-, or Ad-ISF35-treated mice at different effector (E)-to-target (T) ratios was determined by flow cytometry. (F) Splenocytes from vehicle-, Ad5-, and Ad-ISF35-treated mice were cultured with A20 cells, and IFN-γ production was determined by ELISPOT. Statistical analysis was performed using t-test with Welch's correction.