Ischemia‐reperfusion injury (IRI), an innate immunity‐driven local inflammation, remains the major problem in clinical organ transplantation. T cell immunoglobulin and mucin domain (TIM‐3)–Galectin‐9 (Gal‐9) signaling regulates CD4+ Th1 immune responses. Here, we explored TIM‐3–Gal‐9 function in a clinically relevant murine model of hepatic cold storage and orthotopic liver transplantation (OLT). C57BL/6 livers, preserved for 20 h at 4°C in UW solution, were transplanted to syngeneic mouse recipients. Up‐regulation of TIM‐3 on OLT‐infiltrating activated CD4+ T cells was observed in the early IRI phase (1 h). By 6 h of reperfusion, OLTs in recipients treated with a blocking anti‐TIM‐3 Ab were characterized by: (1) enhanced hepatocellular damage (sALT levels, liver Suzuki's histological score); (2) polarized cell infiltrate towards Th1/Th17‐type phenotype; (3) depressed T cell exhaustion markers (PD‐1, LAG3); and (4) elevated neutrophil and macrophage infiltration/activation. In parallel studies, adoptive transfer of CD4+ T cells from naïve WT, but not from TIM‐3 Tg donors, readily recreated OLT damage in otherwise IR‐resistant RAG−/− test recipients. Furthermore, pre‐treatment of mice with rGal‐9 promoted hepatoprotection against preservation‐association liver damage, accompanied by enhanced TIM‐3 expression in OLTs. Thus, CD4+ T cell‐dependent “negative” TIM‐3 costimulation is essential for hepatic homeostasis and resistance against IR stress in OLTs.