Fig. 3.

The mIHC analysis and T/B/NK-cell detection of responders and non-responders. a, b The mIHC results of 1 responded and 1 non-responded tumor. These structures could be visible by staining, including panel I (a): CD8⁺ T cells (green), CD4⁺ T cells (yellow), PD-1⁺ cells (magenta), PD-L1⁺ cells (orange), Foxp3⁺ regulatory T cells (red); and panel II (b): CD19⁺ B cells (green), CD56⁺ NK cells (red), CD68⁺ macrophages (yellow), CD163⁺ M2 macrophages (cyan), cytokeratin⁺ tumor cells (magenta). The proportion of CD8⁺ T cells (c), the ratio of CD8⁺/CD4⁺ T-cell (d), and the proportion of B cells (e) and NK cells (f) in the pre-treatment blood (T1) of responders (red, n = 5) and non-responders (blue, n = 31). The changing trend of immune cells over time (T1–T4) in responders (red, n = 5) and non-responders (blue, n = 31), including the proportion of CD8⁺ T cells (g), the ratio of CD8⁺/CD4⁺ T-cell (h), and the proportion of B cells (i) and NK cells (j). Data (c–f) are shown as means ± SD, with *P < 0.05, **P < 0.01, ***P < 0.001 and ‘ns’ for no significant difference