Fig. 5
Non-canonical MAP-specific T cells in mice rejecting Mlh1-/- tumors. A Immunocompetent mice (BALB/c) were injected with 5X105 Mlh1-/- tumor cells per mouse. Upon rejection, mice were re-challenged twice with Mlh1-/- tumor cells (30 days after the previous injection). Nine days after the last injection, mice were sacrificed, and the spleen were surgically resected. Naïve mice were used as control. B Representative events of viable CD4+ and CD8+ T cells are shown in naïve mice and tumor rejected mice. C Single cells were stained with anti-CD4, CD8, CD44, and CD62L mAb and analyzed by FACS. Data depicts naïve and memory markers of viable CD8+T cells. D Splenocytes were cultured with or without synthesized MAP peptides for 4 days. Viable cells were separated on Ficoll gradients and counted. Total counts are depicted. E After an overnight incubation in IL-2, T cells, from naive and tumor-vaccinated mice, were either restimulated or not with the indicated peptide pools. A scramble peptide (Ctrl) served as control. The supernatant was collected after 48 h to quantify IFN-γ. The data show the release of IFN-γ from splenocytes of individual mice pulsed with scramble or peptide pools. The values of IFN-γ from unpulsed splenocytes were subtracted from the values represented in the figure. p-values were calculated by Mann–Whitney non-parametric test