Figure 2
Flow cytometry analysis to determine HPV16 E7-specific CD8 + T cell responses in tumor-bearing mice treated with bortezomib and/or SAHA. Groups of C57BL/6 mice (5 per group) were challenged with TC-1 tumor cells and treated with bortezomib and/or SAHA according to Figure 1A. Splenocytes from tumor-bearing mice were harvested one week after the last treatment and were stimulated with HPV16 E7aa49-57 peptide (1 μg/ml). The cells were then stained with PE-conjugated anti-mouse CD8a and FITC-conjugated anti-mouse IFN-γ and analyzed by flow cytometry. A. Bar graph depicting the number of E7-specific IFN-γ-secreting CD8+ T cells per 2 × 105 pooled splenocytes (mean + SE; p < 0.01). B. Representative data of flow cytometry E7 peptide-loaded H-2Db tetramer staining in the various groups. C. Peripheral blood mononuclear cells (PBMCs) were harvested from the tail vein one week after the last treatment. Cells were stained with FITC-conjugated anti-mouse CD8a and PE-conjugated HPV16 E7aa49-57 peptide loaded H-2Db tetramer and acquired with FACSCalibur. D. Bar graph depicting the percentage of E7 tetramer CD8+ T cells in PBMCs (p < 0.05).