Figure 4.

Flow cytometric analysis of spleen T cells. The splenocytes were isolated from mice and stimulated with Ags as described in Figure 3. CD4+ and CD8+ T-cell responses were evaluated by using intracellular staining (ICS) for IFN-γ both ex vivo and in vitro by stimulating fresh lymphocytes for 6 h or 72 hours, respectively. Spleen cells from mice of therapeutic vaccination groups challenged with tumor cells and immunized with FSVL and FSWT on day 5, were used for ex vivo (A) as well as for 72 hours (B) ICS assays. Splenocytes from mice used in prophylactic study (as described in Figure 3 B) were assessed for the presence of IFN-γ producing T cells ex vivo (C) or upon stimulation with variant epitopes (D). Data from one representative experiment out of 2 are shown.