Extended Data Fig. 5: Dietary galactose promotes antitumour CD8+ T cell immunity via IGFBP-1.
a, Flow cytometric analysis of proportion of IFN-γ production in CD4+ T cells and CD8+ T cells and exhausted CD8+ T cells treated with 10 mM glucose, 10 mM galactose or 10 mM glucose plus 100 μM galactose (n = 4 biological replicates). CD4+ T cells and CD8+ T cells were isolated from C57BL/6 WT mice. b, Flow cytometric analysis of exhausted CD8+ T cells treated with or without IGF-1 or/and IGFBP-1. c, Flow cytometric analysis of exhausted Igf1r+/+Cd4-Cre and Igf1rfl/flCd4-Cre CD8+ T cells treated with or without IGF-1. d, ELISA analysis of serum IGF-1 from C57BL/6 WT mice fed a high-galactose diet or normal diet at the indicated time points after tumour challenge (n = 5 mice). d, day. e, Immunoblot analysis of the IGFBP-1 proteins in the liver tissue from C57BL/6 WT mice treated with either GalNAc–siIGFBP1 or GalNAc–NC. Liver samples were collected at day 3 after GalNAc–siIGFBP1 treatment. f, Flow cytometric analysis of exhausted CD8+ T cells cultured with medium of hepatocyte from mice with or without GalNAc–siIGFBP1 treatment supplied with or without 10 mM galactose (n = 5 biological replicates). g, Tumour size in C57BL/6 WT mice fed a high-galactose or normal diet and injected s.c. with B16-F10 cells at day 14 after high-galactose treatment. These mice were treated intraperitoneally with 0.03 mg/kg anti-IGFBP-1 or control antibodies (Ctrl Ab) every two days (n = 10 mice) at day 7 after high-galactose treatment. h, Flow cytometric analysis of PD-1 and TIM-3 expression (left, n = 10 mice for Ctrl, n = 8 mice for galactose plus Ctrl Ab and n = 9 mice for galactose plus anti-IGFBP-1) or IFN-γ expression (right, n = 9 mice) in CD8+ T cells from tumours of B16-F10 tumour-bearing C57BL/6 WT mice from g. Tumour-infiltrating CD8+ T cells were isolated at day 14 after tumour injection. The experiments were independently repeated two (a,d–h) or three (b,c) times. Data are presented as mean ± SEM. P values were calculated by one-way ANOVA with Tukey’s multiple comparisons (a,f,h), two-tailed Student’s t-test (d) and two-way ANOVA with Geisser–Greenhouse correction (g). ns, not statistically significant.
