Fig. 3: Systemic anti-tumor immune response induced by localized in vivo T-cell gene-editing.

a Schematic illustration of in vivo T-cell engineering treatment in mice. Tumor volume (b), body weights (c) curves for mice received different treatments as indicated. d Kaplan−Meier plots of the overall survival for animals in different groups. Log-rank tests were performed to analyse the overall survival. For (b−d), n = 5 mice in each group. e Tumor weight of excised tumors at day-19, n = 5 or 10 mice in each group (10 for PD1-CRISPR-Ele and untreated groups, 5 for other groups). Fluorescence microscopy images (f) and fluorescence quantitation (g, h) of the excised tumors after different treatments. Scale bar, 100 µm. Scale bar, 25 µm in the enlarged view, n = 4 independent samples. i Evaluation of tumor-infiltration of CD3⁺, CD4⁺, and CD8⁺ T-cells 19 days after various treatments as indicated, n = 4 mice in each group. The data are presented as mean ± s.e.m. Statistical significance was calculated by two-sided Student’s t-test for two group comparison, one-way ANOVA with Tukey’s test for multiple comparisons. Experimental group information: mice received no treatment (untreated), mice received PD1-CRISPR-Cas9 plasmid in TdLN without electroporation (10 µg, PD1-CRISPR-Diff), mice received intramuscular injection of PD1-CRISPR-Cas9 plasmid (100 µg, IM-DNA-Injection), mice received intravenous injection of anti-PD1 antibodies (20 μg/injection, 3 injections, anti-PD1-Injection), mice received TdLN electroporation of CRISPR/Cas9 plasmid minus gRNA (Null-plasmid-Ele), mice received non-TdLN electroporation of PD1-CRISPR-Cas9 plasmid (10 µg, nonTdLN-PD1-Ele), and mice received TdLN electroporation of PD1-CRISPR-Cas9 plasmid (10 µg, PD1-CRISPR-Ele). Source data are provided as a Source Data file.