Extended Data Figure 9: ICB therapy promotes T_H1 differentiation of CD4⁺-TLs and induces further immune reprogramming.

a, Schematic of the experimental design. b, ICB leads to CD4⁺-TL dependent tumour growth inhibition, measured by tumour weight at Day 15 after E0771 injection. c, d, Total number of immune cells (c) and T cells (d) in tumours from different groups. Although the number of pan tumour-infiltrating immune cells (CD45⁺) is not changed, the number of CD4⁺-TLs increased after immune checkpoint blockade therapy. e, A heat map summarizing changes to tumour-infiltrating immune components after ICB therapy. The number of different immune cells (rows) is shown for each tumour (columns) after control or checkpoint blockade treatment. The weight of each tumour is shown (top panel). Row-side annotations show P values comparing between CD8^KO (anti-IgG) and CD8^KO (anti-PD1 and anti-CTLA4) groups (far left column), and between CD8^KO (anti-PD1 and anti-CTLA4) and TCR^KO (anti-PD1 and anti-CTLA4) (far right column) (EM T, effector memory T cells). f, Quantification of different subsets among CD45⁺CD11b⁺ cells showing the effect of ICB on innate immune microenvironment (eosinophil: CD45⁺CD11b⁺SiglecF⁺). g, h, Quantification of the percentage of T_REG cells among total CD4⁺-TLs, and the ratio of effector memory CD4⁺-TLs to naive CD4⁺-TLs after ICB in CD8 knockout mice. i, Quantification of the percentage of different CD4⁺ T helper cells. j, Percentage of IFNγ⁺ cells in CD4⁺ or CD4⁻ cells among all the CD45⁺ tumour-associated immune cells, indicating CD4⁺-TLs make up the majority of IFNγ⁺ cells. Data are presented as means ± s.e.m. Animal numbers used in (b–j) are denoted in a. P values were calculated using two-tailed unpaired (b–i) or paired (j) Student’s t-test. NS, not significant.