Fig. 4.

The pLegu-H-2Kd minigene vaccine induces IFN-γ release by Legumain specific T-cells (a) Legumain⁺ 4T1 cells freshly harvested from tumor tissues were used as stimulator cells. Flow Cytometry was performed to indicate the extent of Legumain expression on those cells. (b) Production of IFN-γ was verified at the single cell level by ELISPOT as lymphocytes from immunized mice were restimulated with either Legumain⁺ 4T1 tumor cells freshly harvested from tumor tissue or Legumain⁻ 4T1 tissue culture cells. IFN-γ release is indicated by the number of immunospots formed per well. *, P < 0.05, compared with groups of mice whose lymphocytes were not stimulated by Legumain + tumor cells. ^# P < 0.05, compared with control groups. Experiments were repeated three times with similar results