Functional and phenotypic analysis of T cells at tumor site from partial responding or nonresponding patients. (A) T cells, isolated from involved LNs before and after vaccination, were evaluated by IFN-γ ELISPOT assay when cultured alone (□), with the autologous tumor (■), or with an allogeneic tumor (). Results obtained for 2 of 3 partial responders and for a nonresponder patient are reported. Error bars indicate standard deviation of the mean. Statistically significant differences, by 2-sided Student t test, are reported (*P ≤ .05; **P ≤ .01; ***P ≤ .001). (B) Maturation profile of T cells at tumor site before and after vaccination was assessed by flow cytometry analysis of CCR7 and CD45RA expression in the CD3⁺CD8⁺ gated population. Data were acquired using BD CellQuest software version 3.3 on the 4-color analysis flow cytometer BD FACSCalibur. Analyses were performed using FlowJo 8.7.1 software version for Macintosh.