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. 2024 Mar 20;15:2484. doi: 10.1038/s41467-024-46785-9

Fig. 5. Repression of TP63 results in efficient T cell killing and enhances the efficacy of PD-1 mAb therapy in murine SCC models.

Fig. 5

A Flowchart of ex vivo co-culture of murine SCC and OT-I CD8+ T cells. B, C Relative cell viability analysis (B) and crystal violet staining (C) of SCC cells incubation with or without OT-I CD8+ T cells at the indicated effector: target (E: T) ratios. P value for each comparison from left to right in B: 0.1080 (N.S.), 0.0531 (N.S.), 0.0013 (**), 0.0005 (***), 0.0001 (***), 1.88E-05 (***), 0.0988 (N.S.), 0.0846 (N.S.), 0.0059 (**), 0.0249 (*), 5.43E-05 (***), 1.17E-05 (***). D Bright field images showing representative co-cultured MOC22 and OT-I CD8+ T cells. Red arrows indicate a cancer cell killed by activated OT-I CD8+ T cells following 48 hr co-culture (Scale bar, 10 μm). E, F Percent of GZMB and IFNγ production in Scramble or shTrp63 MOC22 (E) and AKR (F) co-cultures. P value for each comparison from left to right in E and F: 0.0467/0.0038, 0.0092/0.0020, 0.0086/0.0072, 0.0281/0.0057. The gating strategy are provided in Supplementary Fig. 6D. G, H Plots of AKR (G) and HNM007 (H) tumor volumes measured every 3 days. C57BL/6J mice were implanted with shTrp63 SCC or Scramble cells and received PD-1 mAb treatment or IgG isotype control (IgG2a). n = 5 for each group. For the tumor volume comparison of Scramble + IgG2a vs. Scramble + α-PD1, Scramble + IgG2a vs. shTrp63 + IgG2a, shTrp63 + IgG2a vs. shTrp63 + α-PD1 and Scramble + α-PD1 vs. shTrp63 + α-PD1 in AKR and HNM007-derived tumor allografts: P = 0.0176/0.0500, P = 0.0014/0.0002, 2.1E-05/.00002, 0.0002/0.0013. (I) Representative IF staining of CD8α of Scramble and shTrp63 AKR allografts, mice were treated with either PD-1 mAb or IgG isotype control (Scale bar, 50 μm). The results were repeated in three biologically independent samples. J, K Tumor growth curves of shTrp63 AKR-bearing (J; P = 0.0001) or HNM007-bearing (K; P = 0.0003) mice treated with PD-1 mAb in combination with CD8 mAb (CD8 T-cell-depletion antibody; n = 5) or IgG isotype control (IgG2b; n = 5). Bars of B–F represent mean ± SD of three biologically independent experiments. P values were determined using a two-sided t -test. The data of (G, H, J, K) were analyzed by a one-sided t -test. N.S. not significant; *P < 0.05, **P < 0.01, ***P < 0.001. Source data are provided as a Source Data file.