Extended Data Fig. 7: Mechanistic studies of NR4A2/IL-12 OT-I T cells and murine anti-hHer2 CAR T cells.

a-e. NR4A2/IL-12 OT-I T cells (CD45.2⁺) were adoptively transferred into congenic CD45.1⁺ mice bearing orthotopic AT-3-ova tumors and ex vivo analyses were performed 8 days post transfer. Transferred OT-I T cells were identified as live CD45.2⁺ Thy1.2⁺ CD8⁺ cells. Endogenous CD8⁺ T cells were identified as live CD45.1⁺ Thy1.2⁺ CD8⁺ cells. a. Quantification of OT-I T cells in the tumor (left) and spleen (right). b. Proportion of splenic OT-I T cells with effector memory (T_EM), central memory (T_CM) or stem cell memory (T_SCM) phenotype, based on CD62L and CD44 expression. Data represent mean proportions from n = 5 mice/group. c. Quantification of PD-1 and Tim3 expression in OT-I T cells in the spleen. d. Concatenated flow cytometry plots (left) and quantification (right) of TNF and IFNγ expression in host CD8⁺ T cells in the dLN. e. Concatenated flow cytometry plots (left) and quantification (right) of Ki67 expression in host CD8⁺ T cells in the tumor. (d-e) Samples were ex vivo stimulated with SIINFEKL peptide for 3 h. (a, c-e) Data represent mean ± SEM from n = 8 (NR4A2/IL-12), 9 (Mock) and 10 (NR4A2 KO) mice/group (tumor), or n = 10 mice/group (spleen) (a), n = 10 mice/group (c), n = 7 (NR4A2 KO), 9 (Mock) and 10 (NR4A2/IL-12) mice/group (d), or n = 8 (NR4A2/IL-12), 9 (Mock) and 10 (NR4A2 KO) mice/group (e) pooled from n = 2 experiments. (d-e) Flow cytometry plots from concatenated samples of n = 5 mice/group. f-g. Mice with previously cured E0771-hHer2 (f) or MC38-hHer2 (g) tumors following treatment with NR4A2/IL-12 murine anti-hHer2 CAR T cells were rechallenged with parental E0771 or MC38 tumors in the opposite mammary fat pad or flank, respectively. Tumor growth curves represented as mean ± SEM from n = 6 mice/group (E0771) or n = 5 (NR4A2/IL-12) and 11 (Naïve) mice/group (MC38). (a, c-e) One-way ANOVA. (f-g) Two-way ANOVA. n.s. not significant, *p < 0.05, **p < 0.01, ****p < 0.0001.

Source Data