(A) Treatment schema. B6 CD90.2 mice were injected intradermally with B16-TMG cells on day –13. On day 0, mice with tumors greater than 2 mm received total-body irradiation and CD90.1 TCX 1.0, TCX 2.0, or negative control cells, followed on day 1 with anti-CD40. Tumor size was followed and measured; mice were euthanized by day 18. (B and C) Both TCX 1.0 and TCX 2.0 cells reduced tumor volume (B) and weight (C). (D) Waterfall plots of change in tumor size from 3 independent experiments combined; mice that died prior to the end of the experiments were censured (see Supplemental Figure 8A). (E and F) TCX 2.0 CD8 cells were more numerous than TCX 1.0 and control cells in spleen (E), whereas there were similar numbers of each recovered from tumor, both greater than control cells (F). (G) More transferred TCX 1.0, TCX 2.0, and control cells were found in the draining LNs (DLNs) than in nondraining cervical lymph nodes (CLNs). (H–J) Dex^OVA MFIs of TCX 1.0 and 2.0 cells were similar in the spleen and DLNs but were greater in tumor-resident TCX 2.0 cells (top panels representative staining; lower panels MFIs from individual mice). (K and L) The dex^OVA MFIs were higher among dex^OVA+ TCX intratumor TCX2.0 cells (K) perhaps due to higher TCRβ expression (L). (M and N) A greater percentage of TCX1.0 cells were TOX⁺ in spleen and tumor (M), whereas a greater fraction of TCX 2.0 cells were CD127⁺ in spleen and DLN (N). Data in B and C are from 1 of 3 experiments with similar results. Differences between groups in B were calculated by determining the AUC of tumor volume versus posttreatment day data for each mouse at the end of the experiment and comparing the groups by ANOVA with Tukey’s post hoc test. P values in C were calculated using a Mann-Whitney rank-sum test. Data in D–N are combined from 3 independent experiments, and groups were compared using a Fisher’s exact test. Significance of cell numbers or MFIs in E–N were determined with Mann-Whitney rank-sum tests.