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. 2012 Nov 23;20(1):57–64. doi: 10.1038/cgt.2012.85

Figure 7.

Figure 7

Long-term functionality of cTCR–CTL and dTCR–CTL in vivo. CTL were purified from pmel-1 or C57BL/6 mice and transduced with anti-flk1 cTCR or anti-flk1 scFv-expressing retroviral vectors. Each CTL was i.v. injected into B16BL6 tumor-bearing mice. As a control, PBS was i.v. injected into some tumor-bearing mice. (a) CTL (2.5 × 106 cells) labeled with PKH26 dyes were i.v. injected into B16BL6 tumor-bearing C57BL/6 mice. The tumor and lymph nodes were removed for preparation of a cell suspension for flow cytometric analysis on day 3 (open square), day 7 (dotted square), day 11 (hatched square) or day 21 (filled square) after CTL injection. Data are presented as mean±s.d. of results from three mice. (b) Three months after injection of 5 × 106 dTCR–CTLs into C57BL/6 mice bearing well-established (∼6 to 6.5 mm in diameter) B16BL6 tumors, CD8-positive T cells were prepared from mice that achieved complete regression of the primary tumor, and then were restimulated with gp10025-33-pulsed dendritic cells for 4 days. The flk1 and gp100-specific cytotoxic activities of these CTL (•) or CTL isolated from C57BL6 (♦) were evaluated by a 51Cr-release assay using MS1 cells, B16BL6 cells, and E.G7-OVA as target cells. Each point represents the mean±s.d. of three independent cultures.