Recipient T cell TIM-3 and hepatocyte galectin-9 signalling protects mouse liver transplants against ischemia-reperfusion injury

J Hepatol. 2015 Mar;62(3):563-72. doi: 10.1016/j.jhep.2014.10.034. Epub 2014 Oct 31.


Background & aims: By binding to T cell immunoglobulin mucin-3 (TIM-3) on activated Th1 cells, galectin-9 (Gal-9) negatively regulates Th1-type alloimmunity. Although T cells contribute to hepatic ischemia-reperfusion injury (IRI), it is unknown whether negative T cell-dependent TIM-3 co-stimulation may rescue IR-stressed orthotopic liver transplants from innate immunity-driven inflammation.

Methods: We used wild type (WT) and TIM-3 transgenic (Tg) mice (C57BL/6) as liver donors and recipients in a clinically-relevant model of hepatic cold storage (20 h at 4°C in UW solution) and syngeneic orthotopic liver transplantation (OLT).

Results: Orthotopic liver transplants in WT or TIM-3Tg→TIM-3Tg groups were resistant against IR-stress, evidenced by preserved hepatocellular function (serum ALT levels) and liver architecture (Suzuki's score). In contrast, orthotopic liver transplants in WT or TIM-3Tg→WT groups were susceptible to IRI. TIM-3 induction in circulating CD4+ T cells of the recipient: (1) depressed T-bet/IFN-γ, while amplifying GATA3 and IL-4/IL-10 expression in orthotopic liver transplants; (2) promoted T cell exhaustion (PD-1, LAG-3) phenotype; and (3) depressed neutrophil and macrophage infiltration/function in orthotopic liver transplants. In parallel studies, we documented for the first time that Gal-9, a natural TIM-3 ligand, was produced primarily by and released from IR-stressed hepatocytes, both in vivo and in vitro. Moreover, exogenous recombinant Gal-9 (rGal-9) potentiated liver resistance against IRI by depressing T cell activation and promoting apoptosis of CD4+ T cells.

Conclusions: Harnessing TIM-3/Gal-9 signalling at the T cell-hepatocyte interface facilitates homeostasis in IR-stressed orthotopic liver transplants. Enhancing anti-oxidant hepatocyte Gal-9 potentiates liver IR-resistance. Negative regulation by recipient TIM-3+CD4+ cells provides evidence for cytoprotective functions of a discrete T cell subset, which should be spared when applying T cell-targeted immunosuppression in transplant recipients.

Keywords: Galectin-9; Innate immunity; Ischemia-reperfusion injury; Liver; Orthotopic liver transplantation; TIM-3.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine
  • Allopurinol
  • Animals
  • Apoptosis
  • Cell Differentiation
  • Disease Models, Animal
  • Galectins / immunology*
  • Glutathione
  • Hepatitis A Virus Cellular Receptor 2
  • Hepatocytes / immunology*
  • Hepatocytes / pathology
  • Immunity, Innate
  • In Vitro Techniques
  • Insulin
  • Liver Transplantation* / adverse effects
  • Lymphocyte Activation
  • Macrophages / immunology
  • Male
  • Mice
  • Mice, 129 Strain
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Mice, Transgenic
  • Neutrophils / immunology
  • Organ Preservation
  • Organ Preservation Solutions
  • Raffinose
  • Receptors, Virus / genetics
  • Receptors, Virus / immunology*
  • Reperfusion Injury / immunology
  • Reperfusion Injury / pathology
  • Reperfusion Injury / prevention & control*
  • Signal Transduction / immunology
  • T-Lymphocytes / immunology*
  • T-Lymphocytes / pathology


  • Galectins
  • Havcr2 protein, mouse
  • Hepatitis A Virus Cellular Receptor 2
  • Insulin
  • Organ Preservation Solutions
  • Receptors, Virus
  • University of Wisconsin-lactobionate solution
  • galectin 9, mouse
  • Allopurinol
  • Glutathione
  • Adenosine
  • Raffinose