Fig. 2.

Frequency of peptide-specific CTLs analyzed by HLA-A24/survivin-2B80-88 peptide tetramer analysis (a case 1, b case 2, c case 7, d case 8 and e case 9). Lymphocytes were collected from peripheral blood of the patients before and after the first, third and sixth vaccinations, stained with an FITC-labeled anti-CD8 mAb and PE-labeled HLA-A24/survivin-2B80-88 peptide tetramer, and analyzed by flow cytometry. As a negative control, a tetramer with an HIV peptide was used. The frequency of CTL precursors was calculated as the number of tetramer-positive cells over the number of CD8-positive cells. The frequencies of HLA-A24/survivin-2B80-88 peptide tetramer -positive CTLs were increased from 0.04 to 1.29%, and 0.07 to 1.34% after the sixth vaccination in cases 1 and 2, respectively (a and b). In case 7, 8 and 9 peptide-specific CTL frequency increased from 0.15 to 0.60%, 0.07 to 0.78% and 0.13 to 1.42%, respectively, after the third vaccination (c–e)