Negative CD4 + TIM-3 Signaling Confers Resistance Against Cold Preservation Damage in Mouse Liver Transplantation

Abstract

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 20h 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 (1h). By 6h 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.

TIM‐3: An emerging target in the liver diseases

“…TIM‐3/Galectin‐9 signalling plays an important role in combating IR‐hepatocyte injury and maintaining OLT homoeostasis. Importantly, overexpression of TIM‐3 by CD4+ T cells can prevent hepatocyte injury in IR stress OLT . Collectively, these data strongly support the CD4+ T cell‐dependent negative TIM‐3 signalling, which is important for maintaining hepatic homoeostasis in related liver damage.…”

“…Isolation of hepatic lymphocytes in an animal model of GVHD, the dysregulation of the TIM‐3 pathway exacerbates the response of GVHD in the liver . Clinical studies suggest that a decrease in the concentration of TIM‐3 in plasma may provide an effective therapeutic measure for future inhibition of GVHD activity and increased tolerance . For note, the TIM‐3/Galectin‐9 pathway negatively regulates T cells in GVHD, and intervention or inhibition of the TIM‐3/Galectin‐9 pathway increased mortality of GVHD.…”

Mesenchymal stem cells improve mouse non-heart-beating liver graft survival by inhibiting Kupffer cell apoptosis via TLR4-ERK1/2-Fas/FasL-caspase3 pathway regulation

“…Afterwards, the liver graft initiates certain mechanisms to repair IRI and hepatocellular damage, and serum ALT/AST levels decrease gradually. However, if IRI is too severe for the graft to repair itself, liver dysfunction occurs within several days, ultimately resulting in recipient death [24–26]. To evaluate liver graft injury, serum and liver specimens were collected and examined at 6 h post-transplantation when inflammation peaked.…”

T Cell Cosignaling Molecules in Transplantation

“…Early following transplantation of cold-stored livers in mice, TIM-3 + CD4 + T cells are observed to infiltrate the graft, and treatment with an anti-TIM-3 blocking antibody resulted in increased histopathologic injury and enhanced hepatocellular damage (Liu et al, 2015b; Liu et al, 2015c). In contrast, adoptive transfer of CD4 + T cells from TIM-3 transgenic donors or pre-treatment of mice with rGal-9 promoted hepatoprotection against preservation-association liver damage in this model (Liu et al, 2015b; Liu et al, 2015c).…”

“…Early following transplantation of cold-stored livers in mice, TIM-3 + CD4 + T cells are observed to infiltrate the graft, and treatment with an anti-TIM-3 blocking antibody resulted in increased histopathologic injury and enhanced hepatocellular damage (Liu et al, 2015b; Liu et al, 2015c). In contrast, adoptive transfer of CD4 + T cells from TIM-3 transgenic donors or pre-treatment of mice with rGal-9 promoted hepatoprotection against preservation-association liver damage in this model (Liu et al, 2015b; Liu et al, 2015c). Interestingly, anti-TIM-3 mAb did not increase hepatocellular damage in TLR-4 deficient animals, suggesting that TIM-3 may function to dampen inflammation that occurs as a result of TLR ligation during the process of tissue preparation, storage, and surgical implantation (Uchida et al, 2010).…”