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. 2014 Aug 20;25(5):277–287. doi: 10.1089/hgtb.2014.004

FIG. 4.

FIG. 4.

Intracellular cytokine production and cytotoxic activity of TCR-transduced T-cells in the GMP-grade cell products. After thawing and overnight recovery, TCR-transduced cells were incubated with HLA-A2+MART-1+ (mel624 and mel526), HLA-A2−MART-1+ (mel938) melanoma cell lines, or cultured alone (−). Intracellular cytokine production was determined after 5 hr of incubation. (A) For T-cells from each validation run, the percentage of IFNγ-, IL-2-, and TNFα-positive CD8+ cells (top row) and CD4+ cells (bottom row) is shown. (B) For the T-cell products incubated with mel624, Boolean gating is shown for CD8+ cells (left panel) and CD4+ cells (right panel). (C) T-cells were taken into culture 1 day before the assay. HLA-A2+MART-1+ (mel526 and mel624) and HLA-A2−MART-1+ (mel938) melanoma lines were loaded with 51Cr and subsequently added to TCR-transduced cells at the indicated effector to target ratios. Lysis was determined after 5 hr of incubation. The percentage of tetramer+ cells of CD3+ cells measured on the day of the assay was 29%, 57%, and 58% for NKITCR001, NKITCR002, and NKITCR003, respectively.