ABSTRACT
BACKGROUND: Pancreatic stellate cells (PSCs) play a critical role in the pathogenesis of pancreatic fibrosis and have emerging functions as progenitor cells, immune cells or intermediaries in pancreatic exocrine secretion. Increasing evidence has shown that desmin as an exclusive cytoskeleton marker of PSC is only expressed in part of these cells. This study was to establish a desmin-positive PSC cell line and evaluate its actions on pancreatic fibrosis, inflammation and immunity. METHODS: The presence of cytoskeletal proteins, integrin α5ß1 or TLR4, was determined by immunocytochemistry while the production of desmin, collagen I, MMP-1, MMP-2, TIMP-2, or CD14 was evaluated by Western blotting. The levels of desmin, collagen I, IL-1 and IL-6 mRNA were determined by real-time quantitative PCR. The secretion of cytokines was detected by ELISA. Cell function was assessed using adhesion, migration, or proliferation assays. RESULTS: A stable activated rat PSC cell line (designated as RP-2) was established by RSV promoter/enhancer-driven SV40 large T antigen expression. RP-2 cells retained typical PSC properties, exhibited a myofibroblast-like phenotype and persistently produced desmin. The cells produced collagen I protein, matrix metalloproteinases and inhibitors thereof. RP-2 cells demonstrated typical PSC functions, including proliferation, adherence, and migration, the latter two of which occurred in response to fibronectin and were mediated by integrin α5ß1. TLR4 and its response genes including proinflammatory cytokines (IL-1, IL-6, TNF-alpha) and chemotactic cytokines (MCP-1, MIP-1α, Rantes) were produced by RP-2 cells and activated by LPS. LPS-induced IL-1 or IL-6 mRNA expression in this cell line was fully blocked with MyD88 inhibitor. CONCLUSION: RP-2 cells provide a novel tool for analyzing the properties and functions of PSCs in the pathogenesis of fibrosis, inflammation and immunity in the pancreas.
Subject(s)
Pancreatic Stellate Cells/metabolism , Pancreatitis/metabolism , Animals , Antigens, Polyomavirus Transforming/genetics , Cell Adhesion , Cell Line, Transformed , Cell Movement , Cell Proliferation , Collagen Type I/genetics , Collagen Type I/metabolism , Desmin/genetics , Desmin/metabolism , Fibrosis , Gene Expression Regulation , Integrin alpha5beta1/metabolism , Interleukin-1/genetics , Interleukin-1/metabolism , Interleukin-6/genetics , Interleukin-6/metabolism , Lipopolysaccharide Receptors/metabolism , Matrix Metalloproteinase 1/metabolism , Matrix Metalloproteinase 2/metabolism , Pancreatic Stellate Cells/immunology , Pancreatic Stellate Cells/pathology , Pancreatitis/immunology , Pancreatitis/pathology , Phenotype , Rats , Respiratory Syncytial Viruses/genetics , Signal Transduction , Tissue Inhibitor of Metalloproteinase-2/metabolism , Toll-Like Receptor 4/metabolismABSTRACT
Type 1 autoimmune pancreatitis (AIP) or chronic sclerosing sialadenitis (Küttner's tumour) is an uncommon disorder that has recently been confirmed as an IgG4-related disease. Here, we describe a rare case of a 53-year-old male patient who primarily presented with pancreatic body mass, left neck mass and several lumps in his lower lip mimicking pancreatic cancer (PC) and neck metastasis. The patient underwent pancreatic body mass and labial gland lumps resection as well as an ultrasound-guided biopsy of the left neck mass. He was diagnosed with IgG4-related focal type of AIP (f-AIP) and Küttner's tumour by immunohistochemistry. The patient responded well to corticosteroid therapy and remains healthy with no signs of recurrence at one year follow-up. The differentiation of f-AIP from PC is very important to avoid unnecessary pancreatic resection.
Subject(s)
Autoimmune Diseases/diagnosis , Head and Neck Neoplasms/secondary , Pancreatic Neoplasms/pathology , Pancreatitis/diagnosis , Sclerosis/diagnosis , Sialadenitis/diagnosis , Submandibular Gland Diseases/diagnosis , Adrenal Cortex Hormones/therapeutic use , Autoimmune Diseases/immunology , Autoimmune Diseases/therapy , Biomarkers/analysis , Biopsy , Diagnosis, Differential , Humans , Immunoglobulin G/analysis , Immunohistochemistry , Male , Middle Aged , Pancreatectomy , Pancreatitis/immunology , Pancreatitis/therapy , Plasma Cells/immunology , Predictive Value of Tests , Sclerosis/immunology , Sclerosis/therapy , Sialadenitis/immunology , Sialadenitis/therapy , Submandibular Gland Diseases/immunology , Submandibular Gland Diseases/therapy , Tomography, X-Ray Computed , Treatment OutcomeABSTRACT
AIM: To determine the utility of connective tissue growth factor (CCN2/CTGF) for assessing hepatic fibrosis in hepatitis B virus (HBV)-induced chronic liver diseases (CLD-B). METHODS: Enzyme-linked immunosorbent assay was used to measure CCN2 in sera from 107 patients with chronic hepatitis B (CHB) and 39 patients with HBV-induced active liver cirrhosis and 30 healthy individuals. Liver samples from 31 patients with CHB, 8 patients with HBV-induced liver cirrhosis and 8 HBV carriers with normal liver histology were examined for transforming growth factor ß-1 (TGF-ß1) or CCN2 mRNA levels by in situ hybridization, and computer image analysis was performed to measure integrated optimal density (IOD) of CCN2 mRNA-positive cells in liver tissues. Histological inflammation grading and fibrosis staging were evaluated by H and E staining and Van Gieson's method. RESULTS: Serum CCN2 concentrations were, respectively, 4.0- or 4.9-fold higher in patients with CHB or active liver cirrhosis as compared to healthy individuals (P < 0.01). There was good consistency between the levels of CCN2 in sera and CCN2 mRNA expression in liver tissues (r = 0.87, P < 0.01). The levels of CCN2 in sera were increased with the enhancement of histological fibrosis staging in patients with CLD-B (r = 0.85, P < 0.01). Serum CCN2 was a reliable marker for the assessment of liver fibrosis, with areas under the receiver operating characteristic (ROC) curves (AUC) of 0.94 or 0.85 for, respectively, distinguishing normal liver controls from patients with F1 stage liver fibrosis or discriminating between mild and significant fibrosis. CONCLUSION: Detection of serum CCN2 in patients with CLD-B may have clinical significance for assessment of severity of hepatic fibrosis.
Subject(s)
Connective Tissue Growth Factor/blood , Hepatitis B, Chronic/complications , Liver Cirrhosis/blood , Liver/chemistry , Adolescent , Adult , Aged , Area Under Curve , Biomarkers/blood , Biopsy , Case-Control Studies , China , Connective Tissue Growth Factor/genetics , Enzyme-Linked Immunosorbent Assay , Female , Humans , In Situ Hybridization , Liver/virology , Liver Cirrhosis/genetics , Liver Cirrhosis/virology , Male , Middle Aged , Predictive Value of Tests , RNA, Messenger/analysis , ROC Curve , Severity of Illness Index , Transforming Growth Factor beta1/genetics , Young AdultABSTRACT
The impact of adefovir dipivoxil (ADV) treatment on the immune system in patients with chronic hepatitis B (CHB) is unknown. The present study was designed to determine the expression of six cytokines, IL-2, IFN-γ, TNF-α, IL-4, IL-6 and IL-10, and their correlation with liver functions and clinical responses to ADV treatment. A total of 22 CHB patients were treated with ADV at a daily oral dose of 10 mg. Six cytokines, as well as AST, ALT and HBV DNA levels in blood samples were quantified prior to and following the treatment. A total of 10 healthy volunteers were enrolled as the control group. The six cytokines in CHB patients were significantly lower than in healthy individuals, and were increased significantly following 4, 12 and 24 weeks of ADV treatment. Although ALT, AST and HBV DNA were reduced following treatment, no correlation was found between these six cytokines and liver function or HBV DNA levels. The levels of the six cytokines in the group of patients with a complete clinical response were significantly higher than those in the group with a partial clinical response. ADV treatment increases the immunity of Th1/Th2 cells in CHB patients, and the increases in cytokines partly reflect the efficacy of the antiviral treatment.
