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IJC_25955_sm_suppinfofig1.tif847KSupplemental Fig. 1. Injection site reaction in patient TK-f02. The reaction after the 3rd (A) and 5th (B) vaccinations of the NY-ESO-1f peptide with Picibanil and Montanide are shown. See text in detail.
IJC_25955_sm_suppinfofig2.tif447KSupplemental Fig. 2. Determination of NY-ESO-1 peptides recognized by CD4 and CD8 T cells in patients. MACS beads-purified CD4 and CD8 T cells (2 × 106) from PBMC obtained at indicated week (w) after vaccination were stimulated once (1°IVS) for 12 days or twice (2°IVS) for 24 days with irradiated autologous CD4- and CD8-depleted PBMC (2 × 106) in the presence of a mixture of 28 18-mer overlapping peptides and a 30-mer C-terminal peptide spanning the entire NY-ESO-1 protein. The cells (4 × 104) from each stimulation culture were then assayed for IFN? secretion by stimulating for 4 hrs with autologous EBV-B cells (4 × 104) in the presence of individual overlapping peptides (peptide 1-29) using FACS. Stimulations with a mixture of OLP and without peptide were included as positive and negative controls, respectively.
IJC_25955_sm_suppinfofig3.tif2925KSupplemental Fig. 3. Clinical responses in patients. A, lung cancer patient TK-f01. A right hilar lymph node (white arrow) and a small metastatic nodule in the left lobe (red arrow) were observed on CT images. The sum of diameters of these target lesions and serum CEA levels were plotted. Vaccination (green triangle) and CT scan (blue triangle) are indicated on a time scale. B, lung cancer patient TK-f02. The sum of target lesion diameters and serum CEA levels were plotted. Metastatic nodule (white arrow) in the right middle lobe was observed on CT images. A small metastatic nodule (red arrow) in the left lower lobe was first noticed in November, 2008. C, esophageal cancer patient TK-f05. Enlarged para-aortic lymph nodes were observed on FDG-PET and CT scan (circumscribed by yellow line). The black arrows indicate the site of vaccination. The red arrow indicates abnormal FDG-uptake in the sternum.

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