Extended Data Fig. 7: Characterization of all intra-tumoural T cells at the therapeutic endpoint.

a, Treatment regimen for the analysis of intra-tumoural T cells at the therapeutic endpoint (mRNA-H 0.25 mg kg⁻¹ and VV-H 2 × 10⁷ pfu per mouse). b, Flow cytometry gating strategy for intra-tumoural exhausted T cells (PD-1⁺TIM-3⁺, PD-1⁺LAG-3⁺ and TIM-3⁺LAG-3⁺). HBsAg specific memory T cells were determined as IFN-γ⁺CD44⁺ cell populations under the stimulation of HBsAg peptides (gate: CD3⁺CD8⁺). c–e. Quantification of intra-tumoural PD-1⁺LAG-3⁺(c), PD-1⁺TIM-3⁺(d) and TIM-3⁺LAG-3⁺ T cells (e) (n = 7 biologically independent samples). f–h. Representative flow cytometry plots of the exhausted T cells, including PD-1⁺TIM-3⁺(f), PD-1⁺LAG-3⁺(g) and TIM-3⁺LAG-3⁺ T cells (h). i,j, Immunohistochemical staining (i) and quantification (j) of intra-tumoural granzyme B secretion (n = 3 biologically independent samples). k,l, Immunohistochemical staining (k) and quantification (l) of intra-tumoural perforin secretion (n = 3 biologically independent samples). For c–e, j and l, data are presented as mean ± s.e.m., with statistical significance assessed using two-tailed Student’s t-test (c–e), and one-way ANOVA (j and l) followed by Tukey’s multiple comparison test.

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