Growth regression of melanoma in NOD/SCID mice treated with TCR gp100-transduced T cells after primary and secondary tumor challenges. (A-B) NOD/SCID mice were engrafted with the MeWo melanoma cell line intradermally (2.5 × 106 cells in 50 μL of matrigel) at the right flank (first tumor cell inoculation, day 0) and adoptively transferred intravenously with T cells transduced with different TCR gp100 constructs (as indicated by symbols) (mean 2.4 × 106 cells in 100 μL of PBS; ADI day 7). (Ai) Effect of ADI on primary tumor outgrowth of animals treated with PBS, Mock, Hu WT TCR gp100, or matrigel. Curves show the cumulative mean and standard error of the mean (SEM) tumor volume of 5 independent experiments (□, ○, ●, ◇). Animals with an intradermal injection of matrigel (50 μL) without tumor cells and animals adoptively transferred intravenously with 100 μL of PBS without T cells were used as controls. P values in brackets indicate the statistical significance of differences in tumor progression over time with the use of a linear regression model with mixed effects (fixed and random) against PBS, Mock. (Bi) Effect of ADI on primary tumor outgrowth of animal treated with PBS, Chim scTCR gp100, Chim scTCR gp100/Mu Cα, and Chim scTCR gp100 S79C/Mu Cα T84C. Curves show the cumulative mean and SEM tumor volume of 5 (□, ▿, ♦) or 2 (▴) independent experiments. The tumor volume of individual mice, their mean, and SEM from experimental groups on day 25 after primary tumor challenge is depicted in panels Aii or Bii. (C-D) Primary local tumors from mice treated with a single injection of TCR gp100-transduced T cells (from A-B) were safely removed after day 25 from the right flank, and 2 days later a secondary tumor inoculation (reset to day 0) with MeWo cell line (2.5 × 106 cells in 50 μL of matrigel) was performed intradermally at the left flank of the same animals. (Ci) Effect of a long-term T-cell response of animal treated with PBS, Mock, Hu WT TCR gp100, or matrigel. Curves show the cumulative mean and SEM tumor volume of 3 (□,○, ◇) or 2 (●) independent experiments. (Di) Effect of a long-term T-cell response of animals treated with PBS, Chim scTCR gp100, Chim scTCR gp100/Mu Cα, and Chim scTCR gp100 S79C/Mu Cα T84C. Curves show the cumulative mean and SEM tumor volume of 3 (□) or 2 (▴, ▿, ♦) independent experiments. The tumor volume of individual mice, their mean, and SEM from experimental groups on day 25 after secondary tumor challenge is given in panels Cii or Dii.
