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. 2016 Dec 14;16(4):782–790. doi: 10.1038/mt.2008.1

Figure 2.

Figure 2

Induction of carcinoembryonic antigen (CEA)-specific cytotoxic T lymphocyte (CTL) by the treatments. (a) Tetramer tests. Representative results derived from triplicate samples were shown. Delayed-type hypersensitivity (DTH) sites–derived emigrated cells were stained with phycoerythrin-labeled HLA-A*0201 tetramers (CAP-1 or SSp-1-specific) and fluorescein isothiocyanate (FITC)-conjugated CD8 monoclonal antibody and finally analyzed by flow cytometry. Numbers indicated for percentages of cells with positive staining. (b) Cytotoxicity assay. DTH site–derived cultured cells were cocultured with 51Cr-labeled SW480 cells, LoVo cells, T2 cells pulsed with CAP-1 (T2/CAP-1), or SSp-1 (T2/SSp-1) peptide. Specific cytotoxicity was evaluated by 51Cr release assay. Results were presented as the mean percentage of specific lysis ± SD of triplicate samples. (c) Interferon-γ (IFN-γ) release assay. CD8+ T cells isolated from the DTH site–derived cultured cells were cocultured with native T2 cells or T2/CAP-1 or T2/SSp-1 for 24 hours. IFN-γ level in the culture supernatant was determined by enzyme-linked immunosorbent assay, and the results were presented as mean ± SD of triplicate samples. Data presented here were the results derived from patient G1 who was treated with 300 μg Aex plus 50 μg granulocyte–macrophage colony-stimulating factor.