Fig 1: IDO1 methylation in distinct leukocytes, HPV-positive, and HPV-negative cell lines. IDO1 methylation at three sites (A: cg10262052, B: cg08465774, C: cg24188163) in isolated leukocytes (N = 52 monocytes, N = 60 B cells, N = 24 CD8+ T cells, N = 94 CD4+ T cells, N = 18 Tregs) from healthy donors, HPV-positive (cg08465774, cg24188163: N = 5; cg10262052 N = 3), and HPV-negative tumor cell lines (cg08465774, cg24188163: N = 34; cg10262052 N = 23). P-values refer to one-way ANOVA. Bars indicate median methylation levels. Results (P-values) from pairwise Bonferroni post-hoc comparisons are listed in Supplemental Table 1.
Fig 2: Kaplan–Meier analysis of overall survival in HNSCC patients stratified according to IDO1 methylation. Patient samples were dichotomized based on an optimized cutoff (69.14% methylation at CpG within the promoter flank targeted by bead cg08465774). Shown are results from N = 527 HNSCC patients from The Cancer Genome Atlas. P-value refers to log-rank test.
Fig 3: Genomic organization of the IDO1 gene. Shown is the IDO1 transcript ENST00000518237.6, CG-density, target sites of HumanMethylation450 BeadChip beads, and the target sequence of the quantitative real-time PCR assay. The modified illustration was exported from www.ensemble.org (Version 89.38) and is based on Genome Reference Consortium Human Build 38 patch release 10 (GRCh38.p10). cg10262052 (bead 1) targets the central promoter site, bead cg08465774 (bead 2) probes the intragenic promoter flank, and bead cg24188163 (bead 3) targets the gene body.
Fig 4: Comparison of overall survival (A) and recurrence-free survival (B) between patients with positive or negative IDO1 expression. Comparison of overall survival (C) and recurrence-free survival (D) between patients with positive or negative PD-L1 expression. Comparison of overall survival (E) and recurrence-free survival (F) between patients with high or low CD8 density.
Fig 5: IDO1 and PD-L1 expression and CD8+ tumor-infiltrating lymphocyte (TIL) status in esophageal squamous cell carcinoma. (A) Positive immunohistochemical staining pattern for IDO1; (B) Negative immunohistochemical staining pattern for IDO1; (C) Positive immunohistochemical staining pattern for PD-L1; (D) Negative immunohistochemical staining pattern for PD-L1; (E) Pattern for high CD8+ TIL density; (F) Pattern for low CD8+ TIL density.
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