(A) Primary data from an exemplary IFN-γ ELISPOT assay. Peripheral blood T cells of 120 patients and 32 healthy donors were stimulated with polypeptides derived from negative control antigen (human IgG), normal self antigens, or the respective test tumor antigens. Data represent mean spot number of 3 wells per antigen ± SEM. **P < 0.05, spot numbers in test wells compared with pooled spot numbers of negative control antigen (2-tailed Student’s t test). TC, T cells. (B) Cumulative total IFN-γ spots from Treg-undepleted T cells for control antigen (IgG) in responding and nonresponding patients and responding and nonresponding TAAs. Mean ± SEM; *P < 0.05 (2-tailed Student’s t test); IgG, n = 21–28; TAAs, n = 29–127. (C) Primary data of an IFN-γ ELISPOT assay with CD45RO⁺ Tmems or CD45RO^– naive T cells. Mean ± SEM; asterisks indicate significant differences between those groups that are connected by horizontal lines. *P < 0.05. (D) Frequency of TAA-specific T cells from responding CRC patients for single TAAs. Mean ± SEM; asterisks indicate significant differences between those groups that are connected by horizontal lines; *P < 0.05; **P < 0.01; n = 2–8. (E) Proportions of CRC patients and healthy donors among all individuals tested that exerted significant T cell reactivity against TAAs from autologous tumor or against polypeptides derived from the respective tumor antigens. Data represent the percentage of patients or healthy donors with TAA-specific T cells; n = 10–36. (F) Number of different TAAs recognized in individual responding patients; n = 25. (G) Frequency of TAA-specific T cells from individual CRC patients and healthy donors (HD). Mean ± SEM. ***P < 0.0001; n = 238–269.