Figure 6. CD8 T cells expanded by vaccines following temozolomide have inferior functional characteristics.

(A) Nur77^GFP TCR signal strength reporter mice (n=4 per group) were treated with the TMZ as indicated and vaccinated daily for 4 days with Ova and poly ICLC. 7 days later splenocytes were plated with the indicated amount of SIINFEKL peptide for 8 hours. Antigen specific T cells were identified by staining for K^b-Ova and CD8, and GFP intensity of antigen specific cells was assessed by flow cytometry. (B) Aggregate data of experiments performed as described in (A). Error bars indicate SEM. *, p<0.05; **, p<0.01; ***, p<0.001. Data shown are pooled from two independent experiments with similar results. (C) C57BL/6 mice (n=3–4 per group) were treated with the TMZ as indicated and vaccinated daily for 4 days with ovalbumin and poly ICLC. 7 days later mice splenocytes were plated for 24 hours with the indicated amount of SIINFEKL peptide. Elaborated IFN-γ was measured by cytokine bead array and normalized to antigen specific T cells as indicated in Materials and Methods. Error bars indicate SEM. *, p<0.05; **, p<0.01. Data shown are pooled data from two independent experiments with similar results.