Targeting TIM-1 on CD4 T Cells Depresses Macrophage Activation and Overcomes Ischemia-Reperfusion Injury in Mouse Orthotopic Liver Transplantation

Abstract

Hepatic injury due to cold storage followed by reperfusion remains a major cause of morbidity and mortality after orthotopic liver transplantation (OLT). CD4 T cell TIM-1 signaling co-stimulates a variety of immune responses in allograft recipients. This study analyzes mechanisms by which TIM-1 affects liver ischemia-reperfusion injury (IRI) in a murine model of prolonged cold storage followed by OLT. Livers from C57BL/6 mice, preserved at 4°C in UW solution for 20h, were transplanted to syngeneic recipients. There was an early (1h) increased accumulation of TIM-1+ activated CD4 T cells in the ischemic OLTs. Disruption of TIM-1 signaling with a blocking mAb (RMT1-10) ameliorated liver damage, evidenced by reduced sALT levels and well-preserved architecture. Unlike in controls, TIM-1 blockade diminished OLT expression of Tbet/IFN-γ, but amplified IL-4/IL-10/IL-22; abolished neutrophil and macrophage infiltration/activation; and inhibited NF-κB while enhancing Bcl-2/Bcl-xl. Although adoptive transfer of CD4 T cells triggered liver damage in otherwise IR-resistant RAG−/− mice, adjunctive TIM-1 blockade reduced Tbet transcription, and abolished macrophage activation, restoring homeostasis in IR-stressed livers. Further, transfer of TIM-1HiCD4+, but not TIM-1LoCD4+ T cells, recreated liver IRI in RAG−/− mice. Thus, TIM-1 expressing CD4 T cells are required in the mechanism of innate immune-mediated hepatic IRI in OLTs.

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

“…Furthermore, the expression of IFN-γ was depressed in TIM-3Tg recipients, along with enhanced GATA-3, IL-4, and IL-10 mRNA levels. A similar Th2-type polarization, with inhibition of Th1/Th17 cytokine programs was detected in liver IRI after TIM-1 blockade [19]. Thus, our findings are in agreement with the immunomodulatory role of TIM-targeted T cell differentiation/function in the early IRI phase in OLT recipients.…”

“…To further elucidate the role of CD4+ T cell-dependent TIM-3 immunomodulation, we used a model of liver IRI in RAG −/− mice [19]. Indeed, adoptive transfer of spleen CD4+ T cells (5×10 6 i.v.)…”

“…into severely T-, B-, and NK T- cell deficient RAG −/− mice (n=3/group). One hour later, RAG −/− test recipients were subjected to 90min of partial local warm ischemia, and liver/sera samples were collected at 6h of reperfusion [19]. …”

“…Standard curves were performed with serial two-fold dilutions; the optical density (OD) was measured with an ELISA reader. Primary mouse hepatocytes, isolated by a two-stage collagenase perfusion method (viability 95–99%), were cultured in complete L-15 medium ± hydrogen peroxide (H 2 O 2 ; 0.4 mM; Sigma-Aldrich) for 4h, and processed by immunofluorescence [19,22]. …”

Associations Between <i>TIM1</i> Polymorphisms and Dilated Cardiomyopathy in a Han Chinese Population

“…Human TIM gene family, which contains TIM1, TIM3 and TIM4, is located on chromosome 5q23-35, a region that has been repeatedly linked with immune dysfunction, which may subsequently lead to autoimmune diseases, allergic diseases, chronic viral infections, immune rejections, and tumors. 10,11,15) The TIM1 gene, also known as hepatitis A virus cellular receptor 1 (HAVCR1) or kidney injury molecule 1 (KIM1), is a vital causative agent of immunological injuries and inflammatory disorders. Previous studies have demonstrated that transient over-expression of TIM1 or cross-linking TIM-1 with agonistic antibodies, in conjunction with T cell receptor (TCR) signal complex and CD28 stimulation, enhances T cell proliferation and cytokine production, which associates with various inflammatory disorders.…”