Fig 2: Clinical outcomes associated with TIM-3 and OX40 expression. TCGA ovarian cancer cohort analysis of mRNA expression (−log10) in (A) OX40 and (B) TIM-3 stratified according to living vs. deceased patients. Transcript expression (−log10) of (C) OX40 and (D) TIM-3 in patients with a low progression-free survival (5–10 months) and high (>45 months) from the TCGA ovarian cancer cohort. Kaplan-Meier survival curve analysis comparing (E) overall and (F) progression-free survival of upper and lower quartile expression of CD4+ OX40+ T cells from Women and Infants Hospital's immunohistochemical data in 29 patients with HGSOC. Log-rank hazard ratios and P-values are reported, with 95% confidence intervals in parentheses. mRNA expression (−log10) of (G) TIM-3 and (H) OX40 grouped according to no macroscopic disease and >20 mm of disease from the TCGA ovarian cancer dataset. (I) % OX40+ CD8+ T cells stratified according to median CA125 preoperative levels obtained from immunohistochemical staining in 29 HGSOC tumors. *P<0.005 as indicated. CA125, cancer antigen 125; NS, not significant; TCGA, The Cancer Genome Atlas.

Fig 3: Comparative ligand expression and pan-cancer analysis for TIM-3 and OX40. TCGA Pan Cancer Atlas Cohort was employed to examine mRNA expression [log2(value+1)] of (A) TIM-3 (B) OX40 and (C) PD-1 across select cancer subtypes. (D) Galectin-9, OX40L and PD-L1 average transcript expression (FKPM) from the TCGA ovarian cancer cohort. TCGA ovarian cancer cohort analysis of (E) OX40L (F) PD-L1 and (G) Galectin-9 mRNA expression (−log10) stratified by median overall survival. *P<0.005 as indicated. NS, not significant; TCGA, The Cancer Genome Atlas; ccRCC, clear cell renal cell carcinoma; pRCC, papillary renal cell carcinoma; PD-1, programmed cell death protein-1.

Fig 4: TIM-3 and OX40 expression in cytotoxic and helper T cell populations. (A) Representative confocal images of the double immunofluorescence staining of TIM-3 or OX40 with CD8 or CD4 from a 29-patient cohort (magnification, ×40). Arrows indicate double-positive cells. (B) Number of CD8+ and CD4+ T cells per field. (C) Number of TIM-3+ and OX40+ cells per field. (D) Average percentages of TIM-3 and OX40 positive CD8+ and CD4+ T cells.