bromochloroacetic-acid and Hepatitis--Chronic

One of the first stages of apoptosis is cytokeratin cleavage mediated by caspases, which is associated with the expression of a neoepitope, the cleavage site of cytokeratin 18, identifiable by the M30 monoclonal antibody. The aim of this study was to evaluate the timing of neoantigen expression and its modifications in the various morphologic stages of apoptosis on frozen and paraffin-embedded sections from liver biopsies of patients with chronic hepatitis or transplanted liver. The appearance of this neoepitope coincides with the gradual disappearance of cytokeratins, with the appearance of nuclear DNA fragmentation, and with the presence of Councilman bodies. The staining patterns on paraffin-embedded sections of liver specimens were similar to those found in frozen sections, with a reduced sensitivity. The M30 antibody is correlated with apoptosis, and its specificity for epithelial cells makes this method the first choice for routine evaluation of apoptosis in liver epithelial cells.

Topics: Antibodies, Monoclonal; Biomarkers; Cell Nucleus; Cytoplasm; DNA Fragmentation; Epitope Mapping; Fluorescent Antibody Technique, Indirect; Frozen Sections; Hepatitis, Chronic; Hepatocytes; Humans; In Situ Nick-End Labeling; Keratins; Liver; Liver Transplantation; Paraffin Embedding

Topics: Adult; Antigens, Differentiation; Bile Ducts, Intrahepatic; Cell Division; Hepatitis, Chronic; Hepatocytes; Humans; Immunohistochemistry; Keratin-7; Keratins; Liver Regeneration; Male; Middle Aged

Topics: Amino Acid Substitution; Biomarkers; fas Receptor; Hepatitis, Chronic; Humans; Keratin-8; Keratins; Liver Failure; Liver Transplantation; Mutation, Missense

Lymphocytic infiltration in the portal triads usually conceals the detection--in haematoxylin and eosin (H&E) stained sections--of bile ducts in two liver diseases: chronic hepatitis and primary biliary cirrhosis. The aim was to assess the number and the characteristics of the bile ducts in those diseases with the aid of an antibody to cytokeratin 7 (CK7).. Consecutive sections from 99 liver biopsies were stained with H&E and anti-CK7.. In H&E sections the total number of central bile ducts in the triads was 52 in primary biliary cirrhosis (n = 37), 69 in chronic hepatitis (n = 43), and 30 in miscellaneous cases (n = 19). Using anti-CK7, the number of central bile ducts was 276 in primary biliary cirrhosis, 348 in chronic hepatitis, and 96 in miscellaneous cases. Central bile ducts with lumen were found in 93.0% of chronic hepatitis cases and in 89.5% of the miscellaneous cases, but in only 13.5% of the primary biliary cirrhosis cases. Peripheral bile ducts in groups of > or = 4/triad were found in all cases of chronic hepatitis (100%) and in 75.7% primary biliary cirrhosis cases, but only in 10.5% of the miscellaneous cases. In 21.6% of primary biliary cirrhosis cases, no bile ducts (central and/or peripheral) were present.. Anti-CK7 detects bile ducts in the triads that are concealed by chronic inflammatory cells. Central and peripheral bile ducts in groups of > or = 4 were significantly more common in primary biliary cirrhosis and chronic hepatitis than in other liver diseases. The lack of lumen in central bile ducts, as well as the absence of central and/or peripheral bile ducts in CK7 stained liver sections, seem to be valuable additional parameters in the differential diagnosis between primary biliary cirrhosis and chronic hepatitis.

Topics: Adult; Aged; Antibodies, Monoclonal; Bile Ducts, Intrahepatic; Biopsy; Diagnosis, Differential; Female; Hepatitis, Chronic; Humans; Immunoenzyme Techniques; Keratin-7; Keratins; Liver Cirrhosis, Biliary; Liver Diseases; Male; Middle Aged

Extracellular matrix (ECM) may affect the function and phenotype of hepatocytes. Phenotypic changes of hepatocytes in diseased liver were investigated with reference to ECM composition.. Immunohistochemistry was performed on biopsied liver samples from chronic viral hepatitis (CVH), primary biliary cirrhosis (PBC), primary sclerosing cholangitis (PSC) and normal patients, using monoclonal antibodies for laminin, type IV collagen, cytokeratin 19 (CK19) and epithelial glycoprotein (EGP), a protein homologous to nidogen.. In normal controls, both EGP and CK 19 were expressed exclusively on biliary epithelia. Laminin and type IV collagen were expressed around portal bile ducts and blood vessels. Although type IV collagen was expressed in Disse's space, laminin was scarcely expressed. In all pathological livers, both EGP and CK 19 were expressed in proliferated bile ductules. In CVH with piecemeal necrosis, EGP was expressed on periportal hepatocytes, while CK19 expression was limited to a few hepatocytes. Laminin was expressed in Disse's space of periportal sinusoids, where EGP was expressed on hepatocytes. EGP expression on hepatocytes and laminin deposition in Disse's space were rare in PBC and PSC liver.. These results suggest that hepatocytes transform into a phenotype similar to biliary epithelia and, laminin deposition in Disse's space (capillarization of sinusoids) may play a role in this phenotypic change.

Topics: Cholangitis, Sclerosing; Collagen; Extracellular Matrix; Hepatitis, Chronic; Hepatitis, Viral, Human; Humans; Immunohistochemistry; Keratins; Liver; Liver Cirrhosis, Biliary; Liver Diseases; Membrane Glycoproteins; Phenotype

An ABC immunohistochemical study of the expression of cytokeratin (CK19 and CK18) was carried out in 315 cases of HBV-caused hepatitis, early-cirrhotic and cirrhotic livers. It was shown that hepatocytes in 73% of chronic active hepatitis (CAH) (80/110) and 81% of early-cirrhotic and cirrhotic livers (117/144) expressed CK19 (the abnormal CK expression), which could be of help in differentiating CAH from chronic persistent hepatitis, subtype CAH (mild, moderate to severe type) and in determining the activity of early-cirrhotic and cirrhotic livers. The abnormal CK expression was shown to be closely related to the activity of liver disorders. The CK19 expression in hepatocytes had the closest relations with the piece-meal necrosis of hepatocytes, isolation of hepatocytes into groups by connective tissue, and fibrosis. It is suggested that CK19 expression may be one of the local reactions to the piece-meal necrosis of hepatocytes.

Topics: Chronic Disease; Hepatitis B; Hepatitis, Chronic; Humans; Immunohistochemistry; Keratins; Liver; Liver Cirrhosis

64 cases of chronic active hepatitis (CAH) and 84 cases of non-CAH liver diseases were studied with keratin stain. The capillary-like bile ductules (CLBD) were proliferating and their morphology was identical to that with Type V collagen stain reported before. CLBD proliferation were more marked in CAH than in other liver diseases, and it was considered to be one of the characteristics of CAH and could be used for differential diagnosis. The ultrastructure of CLBD was specific in morphology. The HBV-DNA in CLBD shown by the technique of in situ hybridization suggested that HBV might infect the cells of CLBD.

Topics: Bile Canaliculi; Hepatitis B; Hepatitis B Core Antigens; Hepatitis B Surface Antigens; Hepatitis, Chronic; Humans; Keratins; Liver

Cholestatic and hepatitic liver cell rosettes, gland-like formations found respectively in chronic cholestasis and in chronic active hepatitis, represent structural modifications of liver cell plates in response to injury. Differences in cytokeratin expression, ultrastructure and three-dimensional (3-D) configuration have been investigated. Cholestatic rosettes are considered to be a form of biliary metaplasia of hepatocytes, linking with newly-formed bile ductules in adjacent septa and probably providing some protection from injury caused by abnormal bile constituents. Hepatitis rosettes, by contrast, are a form of liver cell regeneration developing in isolated surviving hepatocytes or small groups of hepatocytes within areas of collapse.

Topics: Bile Canaliculi; Biopsy; Cholestasis; Hepatitis C; Hepatitis, Chronic; Humans; Immunoenzyme Techniques; Keratins; Liver; Liver Cirrhosis, Biliary; Liver Regeneration; Microscopy, Electron

64 cases of chronic active hepatitis (CAH) and 84 cases of other chronic liver diseases were studied with keratin antibody immunohistochemistry. Proliferation of capillary-like bile ductules (CLBD) was more remarkable in the former cases than that in the latter cases with positive keratin reaction in the cytoplasm. Therefore, proliferation of CLBD was considered to be a characteristic lesion of CAH, which might be used as one of the criteria for differential diagnosis of CAH. This paper also reports the ultrastructural appearance of CLBD in detail and the result of HBV DNA in situ hybridization for CLBD, suggesting that HBV might infect CLBD.

Topics: Animals; Bile Ducts, Intrahepatic; DNA, Viral; Hepatitis B virus; Hepatitis, Chronic; Hyperplasia; Immunohistochemistry; Keratins; Liver; Nucleic Acid Hybridization

Antibodies against thymus epithelial cells (anti-TEC) and the basal cell layer (BCLA) of squamous epithelia have been described in association with HDV-related chronic liver disease (CLD). Data are lacking on their presence during nAnB virus infection. Sera from 51 patients with nAnB post-transfusion hepatitis, including acute and chronic cases diagnosed during a prospective study on candidates for cardiac surgery, and 167 with various forms of CLD were tested for the presence of anti-TEC and BCLA using indirect immunofluorescence on human thymus and rat forestomach sections. Both antibodies mainly occurred in nAnB, HDV and cryptogenic CLD (anti-TEC: 51%, 47% and 42%; BCLA: 29%, 38% and 31%, respectively). The prevalence of anti-TEC in nAnB CLD turned out to be higher than that recorded in alcoholic, HBV-related, autoimmune, liver and kidney microsomal antibody positive CLD and primary biliary cirrhosis (p ranging from less than 0.03 to less than 0.0004). Two monoclonal antibodies (Mabs) to cytokeratins gave a pattern superimposable on that of spontaneous anti-TEC (both Mabs) and BCLA (only one). Antibodies against epithelial constituents, presumably targeting cytokeratin-associated antigens, occur not only in HDV CLD, as previously reported, but also in nAnB CLD, where they might represent a diagnostic aid, due to the unavailability of reliable serological markers of nAnB infection. The close similarity of anti-TEC and BCLA status between nAnB and cryptogenic CLD suggests a nAnB etiology of at least a proportion of chronic liver patients at present scored as cryptogenic.

Topics: Acute Disease; Antibodies, Monoclonal; Antibody Specificity; Autoantibodies; Child; Epithelium; Fluorescent Antibody Technique; Hepatitis C; Hepatitis, Chronic; Hepatitis, Viral, Human; Humans; Keratins; Liver Diseases; Prevalence; Prospective Studies; Thymus Gland; Transfusion Reaction