Fig. 6: NY-ESO-1-specific dTCR-T cells exhibit enhanced in vivo anti-tumor activity in the decitabine-treated AML xenograft mouse model.

A In vivo experimental layout. A total of 3 × 10⁶ fluorescence-expressing U937-A2-luciferase⁺ cells in PBS were s.c. transferred to NCG mice. Tumor growth was monitored by BLI with the depicted regimen. After engraftment was confirmed on day 7, the animals were organized into eight groups according to tumor size (average ~80–100 mm³), including four groups that were treated with PBS (n = 5), NT-T cells (n = 5), TCR-T (n = 5), and dTCR-T cells (n = 5), and another four groups that were treated with DAC alone (n = 5) or in combination with adoptive T cells (NT-T plus DAC, n = 5); TCR-T plus DAC, n = 5); and dTCR-T plus DAC, n = 5). DAC was given on days 7–11 via i.p. injection at a dose of 1.0 mg/kg body weight in 100 μL of PBS. On day 13, mice (except for PBS and DAC groups) were treated by i.v. injection of 1.0 × 10⁷ NT-T cells, TCR-T cells, or dTCR-T cells. Tumor volume was assessed every 3 days using calipers with the formula (length × width²)/2) and weekly by BLI with the IVIS Lumina III System. B Western blot analysis of the expression of NY-ESO-1 protein in tumor tissues 3 days after the last administration of DAC, with the multiple myeloma cell line U266 as the positive control (n = 3). C Mean tumor growth curves of the eight groups (n = 5). D Qualification of the BLI signal from each treatment group over the time (n = 5). Data are represented as the total fux (photons/second). E BLI images showing tumor burdens in all mice at the indicated time points (n = 5). F Kaplan–Meier survival curves presenting the overall survival of each group (n = 5). The log-rank Mantel–Cox test was used to analyze the survival of each group.