Extended Data Fig. 3. Orthogonal ACT expands CD8+ TILs in situ in a cell autonomous manner.
a General gating strategy to analyze the adoptively transferred OT1 TILs. Mice with B16.OVA tumors were treated with either engineered or untransduced OT1 cells on days 12 and 15 after tumor cell inoculation; then tumors were harvested on days 5 and 12 post ACT and cell quantification was performed by flow cytometry using precision counts beads. b Total number of CD8+ TILs at day 5 in mice treated with orthogonal ACT versus non-treated tumor bearing mice (17 days post tumor inoculation) evaluated in parallel. Data are from three independent experiments (n = 5 mice/group). Data are presented as mean values ± s.d. A two-tailed Student’s t-test with Welch’s correction was used for comparing both conditions****P < 0.0001. c Intratumoral persistence of OT1 TILs at day 12 post ACT in each studied group. Right: representative dot plots for each treatment condition. Data are from 4 independent experiments (n = 4−5 animals/group/experiment). Data are presented as mean values ± s.d. An ordinary one-way ANOVA with a Tukey’s test for multiple comparison was performed for comparing conditions differences, *P < 0.05, **P < 0.01 ***P < 0.001, ****P < 0.0001. d Pearson’s correlation between the numbers of intratumoral TCF1+ OT1 T cells and the total number of intratumoral OT1 T cells on day 12 post orthogonal ACT. Data are from two independent experiments (n = 6 animals/group/experiment).