lipoteichoic-acid and Liver-Cirrhosis--Biliary

Previous studies have shown differential TIPE2 expression in several autoimmune diseases. However, the expression levels of TIPE2 in primary biliary cirrhosis (PBC) remained unclear. The purposes of this study were to evaluate TIPE2 expression levels in patients with PBC and further investigate its role in PBC pathogenesis.. A total of 40 PBC patients and 44 healthy controls were included in the present study. Quantitative reverse-transcription polymerase chain reaction and western blotting were used to determine the differences in mRNA and protein expression levels of TIPE2. The correlations of TIPE2 expression levels and clinical characteristics, inflammatory cytokines, and ursodeoxycholic acid treatment were also assessed. Besides, the influence of TIPE2 on the reactivity of monocyte to Toll-like receptor ligands was further analyzed.. The expression levels of TIPE2 were significantly decreased in PBC patients compared with normal controls (P < 0.01). The expression levels of TIPE2 were negatively correlated with alanine aminotransferase (r = -0.40, P = 0.01), alkaline phosphatase (r = -0.36, P = 0.02), gamma glutamyl transpeptidase (r = -0.53, P < 0.01), tumor necrosis factor (TNF)-α (r = -0.332, P = 0.03), interleukin (IL)-1β (r = -0.386, P = 0.01), and IL-8 (r = -0.366, P = 0.02) levels in sera from PBC patients. TIPE2 expression level could be significantly increased after ursodeoxycholic acid treatment (P < 0.01). The production of TNF-α, IL-1β, and IL-8 by monocytes from PBC patients after stimulation with lipopolysaccharide and lipoteichoic acid was significantly increased when TIPE2 was knocked down. Furthermore, TIPE2 knockdown could promote activation of nuclear factor-κB pathways through increasing phosphorylation and degradation of IκB in peripheral blood monocytes from PBC patients.. The present study reported that insufficient expression of TIPE2 might be involved in the hyperreactivity of monocyte to Toll-like receptor ligands in PBC.

Topics: Adult; Case-Control Studies; Cells, Cultured; Cholagogues and Choleretics; Cytokines; Down-Regulation; Female; Humans; Inflammation Mediators; Intracellular Signaling Peptides and Proteins; Ligands; Lipopolysaccharides; Liver Cirrhosis, Biliary; Male; Middle Aged; Monocytes; NF-kappa B; RNA Interference; RNA, Messenger; Signal Transduction; Teichoic Acids; Toll-Like Receptors; Transfection; Ursodeoxycholic Acid

Chronic colitis-harboring TCRalpha(- / - ) x AIM(- / - ) mice showed PBC-like bile duct damage in the liver. Bacterial infection is one of the candidates for the pathogenesis of PBC. We demonstrated that the bacterial cell wall component lipotheicoic acid (LTA) was detected at sites of inflammation around damaged bile ducts in PBC patients. The aim of this study was to investigate the pathophysiology of the liver and other organs in TCRalpha(- / - ) x AIM(- / - ) mice.. Thirteen female TCRalpha(- / - ) x AIM(- / - ) mice were sacrificed at 24 weeks of age. The liver, stomach, small intestine, colon, pancreas, kidney and spleen were studied for pathological examination. Using anti-LTA antibody as the primary antibody, an immunohistochemical study was carried out.. In the liver, LTA was mainly detected in the portal area with inflammation, and some of the cytoplasm of hepatocytes. Inflammations were also observed in the stomach, intestine, pancreas and kidney. Throughout the gastrointestinal tract, from the stomach to the colon, LTA was detected in the epithelium at sites of inflammation. Furthermore, LTA was detected around both pancreatic ducts with inflammation and distal renal tubules with inflammation.. The development of inflammations in the liver as well as extensive organs, strongly suggests a close relationship between bile duct damage and systemic multifocal epithelial inflammations, perhaps involving bacterial LTA, in TCRalpha(- / - ) x AIM(- / - ) mice.

Topics: Animals; Bile Ducts, Intrahepatic; Chronic Disease; Colitis; Epithelium; Female; Gram-Positive Bacteria; Inflammation; Intestine, Small; Kidney; Lipopolysaccharides; Liver; Liver Cirrhosis, Biliary; Mice; Pancreas; Spleen; Teichoic Acids

Autoimmune disorder and associated multifocal organ inflammations such as dry gland syndrome are occasionally observed; however, their etiologies are not clearly understood. We previously reported that chronic colitis-harboring TCR alpha(-/-) x AIM(-/-) mice show primary biliary cirrhosis (PBC)-like bile duct damage in the liver. Gram-positive bacterial infection is one of the candidates for the pathogenesis of PBC. We also reported that the bacterial cell wall component lipoteichoic acid (LTA) was detected at the sites of inflammation around damaged bile ducts in PBC patients. On the basis of these facts, we hypothesized that LTA might affect the pathogenesis of bile duct damage in the livers of TCR alpha(-/-) x AIM(-/-) mice. LTA was detected not only in the portal area with inflammation in the liver but also throughout the gastrointestinal tract, from the stomach to the colon, and especially in the epithelium at sites of inflammation. In addition, LTA was detected around both pancreatic ducts with inflammation and at the distal renal tubules with inflammation in TCR alpha(-/-) x AIM(-/-) mice. Furthermore, in the liver, pancreas, kidney, and colon, fibrous stroma were detected at the sites of LTA-positive inflammation foci. Bacterial LTA might affect the pathogenesis of epithelial inflammation followed by fibrosis in systemic multifocal epithelial inflammations in chronic colitis-harboring TCR alpha(-/-) x AIM(-/-) mice with PBC-like bile duct damage.

Topics: Animals; Apoptosis Regulatory Proteins; Colitis; Epithelium; Fibrosis; Inflammation; Lipopolysaccharides; Liver Cirrhosis, Biliary; Mice; Mice, Knockout; Receptors, Antigen, T-Cell, alpha-beta; Receptors, Immunologic; Receptors, Scavenger; Teichoic Acids

Intrahepatic bile ducts are the targets for inflammation in primary biliary cirrhosis (PBC), but their pathogenesis is not known. Gram-positive bacterial DNA was detected recently in gallbladder bile of PBC patients. In the present study, we assessed the possible pathological role of lipoteichoic acid (LTA), the gram-positive bacterial cell wall component, in PBC.. Liver samples, obtained from 20 patients with PBC (stage 1-2 with CNSDC: stage 3-4 with loss of bile ducts = 10:10) and from 13 patients with chronic hepatitis due to hepatitis C virus (CH-C) with lymphocytic cholangitis, were subjected to immunohistochemical staining with polyclonal rabbit anti-LTA as the primary antibody. Serum reactivities to LTA were studied by ELISA. After 1 microg of purified LTA was placed in a 96-well microplate as an antigen, an antibody capture assay was carried out using serum samples from PBC (n = 20), CH-C (n = 13) and healthy subjects (n = 11).. LTA was localized around the sites of chronic non-suppurative destructive cholangitis (CNSDC) in the portal area in stage 1-2 PBC but was not detected in the portal area in CH-C. In stage 3-4 PBC, LTA was localized around sites of ductular proliferation at the periphery of portal tracts. IgM class anti-LTA serum titers were significantly higher in PBC than in CH-C. IgA class anti-LTA serum titers were significantly higher in PBC than in healthy subjects.. In the PBC livers, the profile of immunoreactivity to LTA changed markedly as the disease progressed. Sera from PBC showed higher levels of anti-LTA titers than CH-C (IgM) or from healthy subjects (IgA). The LTA-mediated immune system might affect the initiation and/or progression of PBC.

Topics: Adult; Aged; Antibodies, Bacterial; Bile Ducts, Intrahepatic; Female; Gram-Positive Bacteria; Hepatitis C, Chronic; Humans; Lipopolysaccharides; Liver Cirrhosis, Biliary; Middle Aged; Teichoic Acids

The role of the adaptive immune response, with regard to the development of autoantibodies, has been extensively studied in primary biliary cirrhosis (PBC). However, the importance of innate immunity has been noted only recently. Based on the proposed role of microorganisms in the pathogenesis of the disease, we hypothesize that patients with PBC possess a hyper-responsive innate immune system to pathogen-associated stimuli that may facilitate the loss of tolerance. To address this issue, we isolated peripheral blood monocytes from 33 patients with PBC and 26 age-matched healthy controls and stimulated such cells in vitro with defined ligands for toll-like receptor (TLR) 2 (lipoteichoic acid; LTA), TLR3 (polyIC), TLR4 (lipopolysaccharide; LPS), TLR5 (flagellin), and TLR9 (CpG-B). Supernatant fluids from the cultures were analyzed for levels of 5 different pro-inflammatory cytokines, interleukin (IL)-1beta, IL-6, IL-8, IL-12p70, and TNF-alpha. After in vitro challenge with TLR ligands, PBC monocytes produced higher relative levels of pro-inflammatory cytokines, particularly IL-1beta, IL-6, IL-8, and TNF-alpha, compared with controls. In conclusion, monocytes from patients with PBC appear more sensitive to signaling via select TLRs, resulting in secretion of selective pro-inflammatory cytokines integral to the inflammatory response that may be critical in the breakdown of self-tolerance.

Topics: Adult; Aged; Cytokines; Female; Flagellin; Humans; In Vitro Techniques; Ligands; Lipopolysaccharide Receptors; Lipopolysaccharides; Liver Cirrhosis, Biliary; Male; Membrane Glycoproteins; Middle Aged; Monocytes; Oligodeoxyribonucleotides; Poly I-C; Receptors, Cell Surface; Teichoic Acids; Toll-Like Receptor 2; Toll-Like Receptor 3; Toll-Like Receptor 4; Toll-Like Receptor 5; Toll-Like Receptor 9; Toll-Like Receptors

Gram-positive bacterial DNA is frequently detectable in gallbladder bile of primary biliary cirrhosis (PBC) patients. To advance these findings, lipoteichoic acid (LTA) of gram-positive bacteria with high antigenicity was examined in liver specimens and bile from PBC patients and controls.. LTA was examined by Western blotting in the gallbladder bile from 15 PBC, 11 cholecystolithiasis and six normal subjects, and by immunohistochemistry in liver specimens from 16 PBC, six primary sclerosing cholangitis (PSC), eight chronic viral hepatitis C (CVH-C) and five normal subjects.. In the gallbladder bile, there was no significant difference in the positive rate of LTA between PBC and controls. LTA-containing mononuclear cells were frequently detected in the portal tracts, particularly around the bile ducts and in hepatic sinusoids in PBC, while they were infrequent or occasional in control livers. These LTA-containing cells were sinusoidal endothelial cells and Kupffer cells, and portal monocytes, which frequently expressed scavenger receptor class B type 1.. LTA derived from bacterial fragments may reach the bile, not only in the diseased state but also under normal conditions. Such LTA may be involved in the development and progression of portal tract lesions, particularly bile duct lesions, in PBC.

Topics: Adult; Aged; Bile; Bile Ducts, Intrahepatic; Case-Control Studies; CD36 Antigens; Female; Gram-Positive Bacteria; Granuloma; Humans; Immunohistochemistry; Lipopolysaccharides; Liver; Liver Cirrhosis, Biliary; Male; Membrane Proteins; Microscopy, Immunoelectron; Middle Aged; Receptors, Immunologic; Receptors, Lipoprotein; Receptors, Scavenger; Scavenger Receptors, Class B; Teichoic Acids