Fig. 5: The combination of an ALKi and anti-PD-1 therapy leads to macrophage polarization reprogramming and enhancement of immune effector functions.

a Schematic of the protocol for combination treatment with αPD-1 and crizotinib using an orthotopic HCC model. Figure was created with BioRender.com. b Magnetic resonance images of orthotopic liver tumors in the indicated groups of mice on day 5 (D5) and day 25 (D25) after tumor-cell implantation. c Mouse tumor weights measured at the treatment endpoint (n = 5 per group). d–f Percentages (d) and absolute numbers (e, f) of CD206⁺ and CD206⁻ TAM subsets in the tumors. g, h Absolute numbers of CD8⁺ T cells (g) and IFN-γ⁺granzyme B⁺ CD8⁺ T cells (h) in the tumors. i, j Absolute numbers of Th1-cell (i) and Treg (j) subsets in the tumors. For e–j n = 5 for control group; n = 6 for anti-PD-1 or Crizotinib treatment group; and n = 7 for combination group. k OS durations in mice bearing HCA-1-RNase1 tumors following treatment, as indicated (n = 6 mice per group). Log-rank test. l IHC staining from tumors of indicated group. Scale bar, 50 µm. m Quantification of IHC score of CD206, Granzyme B and PD-L1 in indicated groups (n = 5 per group). n, o The tumor growth of Hepa1-6 cells in C57BL/6 mice treated with Alectinib, anti-PD-1, or the combination. Representative images of tumors are shown on (n) and quantification of tumor volume is shown on (o). n = 6 mice per group. p–r Absolute numbers of iNOS⁺ CD206⁻ TAM (p), CD8⁺ T cells (q) and IFN-γ⁺ GranB⁺ CD8⁺ T (r) cells in the tumors (n = 6 per group). a–k Representative results from 2 independent experiments. n–f Representative results from 3 independent experiments. The error bars represent mean ± SD values. Statistical analysis: c, e–j, m, and p–r, two-sided Unpaired Student’s t-test. o a log-rank test. Source data are provided as a Source Data file.