Immunologic and clinical responses of patient no. 1 with acute myeloid leukemia to the course of RHAMM-R3 peptide vaccination. (A) The FACS dot plots show the percentage of HLA-A2/R3-tetramer⁺/CD8⁺ T lymphocytes. The frequency of CD8⁺/HLA-A2/R3-tetramer⁺ T lymphocytes increased after 4 vaccinations (i represents before; ii, after second; iii, after third; and iv, 2 weeks after fourth/last vaccination). The HLA-A2/R3-tetramer*PE–positive CD8⁺ T cells were further analyzed for their expression of CCR7 and CD45RA. Most of the cells (70%-85%) revealed to be CD8⁺/HLA-A2/R3 tetramer⁺/CCR7⁻/CD45RA⁺ effector T cells. (B) The frequency of R3-specific CD8⁺ T lymphocytes increased during the course of vaccination as assessed by ELISpot assays for IFN-γ and granzyme B release. All assays were performed in triplicate. Error bars indicate the SD. (C) Monitoring of malignant cells correlated with the other findings showing a decrease of CD33⁺/HLA-DR⁺ malignant cells during vaccination. Numbers on plots are percentages of all lymphocytes (panel A), or of cells in the blast gate (panel C), respectively.