Long term immune responses of patient 3 showing durable disease stabilitazion. a. Lymphoproliferation assay in response to antigen stimulation of PBMCs from pt3 collected at T21m, compared with results obtained from the previous tests. b. Lymphocyte cultures (the same as in panel a) analyzed for cytokine production by Bio-Plex® Multiplex System. Only cytokine-positive antigen-specific cultures are shown. Negative controls showed no detectable cytokine levels except for IL-8-specific samples (2.6 × 103 pg/ml and 3 × 103 pg/ml for, respectively, time points T0 and T21). c. Immunofluorescence of biopsy from pt3 metastatic lesion. The tumor sample was collected 1 month after the last evaluation of patient clinical status by PET (T21m). The immunofluorescence microscopy was done using antibodies specific for CD45, CD45RO, and CD68 stained with 4′,6-diamidino-2-phenylindole (DAPI). Images show that tumor lesion is abundantly infiltrated both by T lymphocytes (a) which are characterized by a memory phenotype (b) and a monocyte/macrophage population (c).