The study protocol conformed to the ethical guidelines of the 1975 Declaration of Helsinki; this was reflected in a priori approval by the local ethics committee, and written, informed consent was obtained from each patient.17 As controls, healthy volunteers with normal aminotransferase activity, no history small molecule library screening of liver disease or alcohol abuse, and negative serology for hepatitis B virus, hepatitis C virus, and human immunodeficiency virus were recruited from the local blood transfusion institute. Samples from 30 liver biopsy

procedures and 33 explanted cirrhotic livers (staged by a blinded pathologist) were subjected to messenger RNA (mRNA) expression analysis. Fractalkine serum concentrations were measured with a cytometric bead array (#560265, BD Biosciences). C57BL/6 wild-type (WT), congenic Ly5.2 (CD45.1) C57BL/6, and CX3CR1−/− mice (backcrossed for more than eight generations to a C57BL/6 background) were

maintained in a pathogen-free environment. The CX3CR1-deficient mice had green fluorescent protein (gfp) inserted into the CX3CR1 genetic locus.10 Experiments were performed with age-matched and sex-matched animals at 6 to 8 weeks of age. selleck kinase inhibitor Mice were treated with carbon tetrachloride (CCl4; Merck; 0.6 mL/kg of body weight) Ureohydrolase mixed with corn oil or were treated with corn oil (for controls). Mice were sacrificed 48 hours after the last injection. Bile duct

ligation (BDL) was performed according to standard procedures. All animals received humane care, and the experiments were approved by appropriate German authorities. The isolation of blood cells and intrahepatic leukocytes was performed,5 and they were then subjected to fluorescence-activated cell sorting (FACS) with the following monoclonal antibodies: F4/80 (Serotec); CD45, Ly6G, natural killer 1.1, CD3, and CD8 (BD); and CD115, CD4, CD11b, CD45.1, CD45.2, and CD11c (eBiosciences). For the analysis of intrahepatic macrophage apoptosis, intrahepatic leukocytes were stained with CD45, CD11b, F4/80, and annexin V/allophycocyanin (APC; BD). CCR2 staining was performed with an unconjugated antibody (Epitomics) or with a corresponding isotype control (rabbit immunoglobulin G; BD), which was followed by secondary goat anti-rabbit APC (Invitrogen).