摘要:

AbstractRecent studies have demonstrated the presence of hepadnavirus‐related nucleic acids in extrahepatic tissues in various animal models. The prevalence and biological significance of extrahepatic infection in humans remains undetermined. To characterize the tissue distribution and physical state of extrahepatic hepatitis B virus nucleic acids in acute hepatitis infection, we examined serum, liver and multiple extrahepatic tissues obtained at autopsy from two patients with fulminant hepatitis and one patient with resolving hepatitis who died of an unrelated cause. Southern‐blot hybridization analysis was used to analyze the physical state of hepatitis B virus‐related DNA. Hepatitis B virus‐related RNA sequences were examined by slot‐blotting total RNA extracted from corresponding tissues. Hepatitis B virus nucleic acids were demonstrated in lymph nodes, spleen, gonads, thyroid gland, kidneys, pancreas and adrenal glands. The most intense signal of hybridization was obtained with DNA extracted from lymph nodes. In general, the levels of hepatitis B virus RNA correlated with the amount of viral DNA. Fast‐migrating DNA sequences resembling replicative intermediates and ranging in size from 1 to 3.2 kb were detected inEcoRI digests. Faint high‐molecular‐weight smears suggesting random integration also were observed. Remarkably, little or no hepatitis B virus nucleic acid was detected in the serum or liver. In control specimens obtained from hepatitis B virus carriers, most hybridizable hepatitis B virus nucleic acid was present in liver, but hepatitis B virus DNA was also detected in extrahepatic tissues. Finally, no specific histological changes were observed in extrahepatic tissues harboring hepatitis B virus. These findings demonstrate hepatitis B virus nucleic acids in extrahepatic sites in humans during acute infection and provide new insights into the pathogenesis of fulminant hepatitis. (HEPATOLOGY1

ISSN:0270-9139    

DOI:10.1002/hep.1840120202

出版商:W.B. Saunders    

年代:1990

数据来源: WILEY

摘要:

AbstractWe have studied antibodies (anti‐pol antibody) against the polymerase gene product of hepatitis B virus by solid‐phase enzyme immunoassay using synthetic peptides coded for by this gene. Sera from six patients with acute hepatitis B, 112 chronic hepatitis B virus carriers and six healthy individuals with naturally acquired immunity to hepatitis B virus were tested for anti‐pol antibody. In acute hepatitis B virus infection, anti‐pol antibody was detected in three of six patients. In chronic hepatitis B virus infection, antipol antibody was detected in 17 of 29 (59%), in 23 of 33 (70%) of cirrhotic patients and in 18 of 24 (75%) patients with cirrhosis complicated by hepatocellular carcinoma, compared with 4 of 19 (21%) asymptomatic carriers and 2 of 7 (29%) patients with chronic persistent hepatitis. Titers of anti‐pol antibody were higher in cirrhotic patients with and without hepatocellular carcinoma than in patients with chronic active hepatitis. The presence of anti‐pol antibody, however, had no relationship with hepatitis B virus‐associated DNA polymerase activities and other viral replicative markers. As for sera from six healthy individuals with naturally acquired immunity to hepatitis B virus, two (33%) were positive for anti‐pol antibody. These results indicate that the immune response toward the polymerase gene product is induced during acute and chronic hepatitis B virus infection. In chronic hepatitis B virus infection, anti‐pol antibody may serve as a new marker indicative of a long period of hepatitis B virus‐induced hepatitis. (HEPATOL

ISSN:0270-9139    

DOI:10.1002/hep.1840120203

出版商:W.B. Saunders    

年代:1990

数据来源: WILEY

摘要:

AbstractThe preS1 domain of hepatitis B virus envelope proteins contains a site of attachment to the hepatocyte membrane that has been shown to evoke virusneutralizing antibodies. Using synthetic peptides, we have examined kinetics and specificity of the antibody response to preS1 during acute and chronic HBV infection. Antibodies against two continuous B cell epitopes, p (21–32) and p (32–47), which overlap with the virus receptor for hepatocytes, were detected in 17 (28%) and 28 (47%) patients, respectively, of 60 patients who were tested during acute hepatitis B. Serial testing demonstrated these anti‐preS1 specificities in more than 50% of patients who became virus free. By contrast, five patients with chronic evolution of hepatitis B and 61 of 66 patients with an established chronic HBV infection were negative, independent of serological profile and liver disease activity. Fifteen (22.7%) patients with chronic hepatitis B were positive for antibody to the C‐terminus p (94–117) preS1 sequence that, unlike the acute‐phase anti‐(21–32) and anti‐(32–47) reactivities, did not behave as a virusprecipitating antibody. Acute‐phase sera were found to also contain virus‐precipitating antibodies directed against conformational preS1 epitopes. These results indicate that the preS1 site, which contains the binding activity for the hepatocyte membrane, elicits an early antibody response during acute hepatitis B. A defect in such antibody repertoire may participate in the chronicity process as a result of continuing reinfection of hepatocytes by circulating virions. (HEP

ISSN:0270-9139    

DOI:10.1002/hep.1840120204

出版商:W.B. Saunders    

年代:1990

数据来源: WILEY

摘要:

AbstractWe have developed a rapid method to characterize genomic diversity of low‐level hepatitis B and related viral agents after their identification in serum by high‐affinity HBsAg‐antibody monoclonal antibody capture and subsequent polymerase chain reaction amplification. Serum from an individual with chronic liver disease and without hepatitis B virus serological markers but reactive by monoclonal antibody capture/polymerase chain reaction amplification was inoculated into a chimpanzee. After inoculation, an acute hepatitis B virus‐like hepatitis developed in the chimpanzee. Analysis of serial liver biopsy samples showed the persistence of hepatitis B virus DNA for more than 17 mo after resolution of acute hepatitis and seroconversion. Applying the technique of restriction enzyme fragment analysis to serial chimpanzee liver biopsy samples and acute‐phase sera, along with the serum inoculum, we established that all hepatitis B virus DNA sequences are derived from the same viral agent. We present evidence that the viral DNA persisted as a nonreplicating episomal form in the nucleus of hepatocytes. This study demonstrates that after clinical and serological recovery from an acute hepatitis, there may be persistence of low‐level hepatitis B virus‐related genome in the liver despite the presence of antibodies to HBsAg. Such persistence of viral genome may be a natural sequela of infection and may serve as a source of viral latency and possible reactivation. Finally, cloning and complete nucleic‐acid sequencing of this virus have demonstrated multiple nucleotide and amino acid changes compared with all known hepatitis B virus subtypes. These changes may have contributed in part to a different antigenic composition or immunological reactivity of the host to this hepatitis B virus isolate. (HEPATOLOGY1

ISSN:0270-9139    

DOI:10.1002/hep.1840120205

出版商:W.B. Saunders    

年代:1990

数据来源: WILEY

摘要:

AbstractThe plasma disappearance rate of sulfobromophthalein (VBSP; μmol/kg/min) was measured in 15 Gilbert's syndrome patients and 12 control subjects after intravenous injection of two different doses (0.59 and 5.90 μmol/kg) of the dye. Plasma disappearance rate was significantly reduced in Gilbert's syndrome patients after administration of 0.59 μmol sulfobromophthalein/kg (0.119 ± 0.016 vs. 0.146 ± 0.018 μmol/kg/min; mean ± S.D.; p<0.001), whereas no difference was found with the higher dose (0.754 ± 0.040 vs. 0.767 ± 0.072 μmol/kg/min). Significant reduction was also found after administration to four Gilbert's syndrome patients and four control subjects of 0.29 and 2.95 μmol sulfobromophthalein (0.060 ± 0.005 μmol/kg/min vs. 0.077 ± 0.07 μmol/kg/min and 0.480 ± 0.012 μmol/kg/min vs. 0.591 ± 0.015 μmol/kg/min, respectively; p<0.01). Competition studies with combined administration of sulfobromophthalein (0.59 μmol/kg) and different doses of rifamycin SV (0.59, 1.47 and 2.95 μmol/kg) showed a significant (p<0.001) reduction in plasma disappearance rate in Gilbert's syndrome patients but not in controls. The rifamycin SV dose at which a 50% inhibition in plasma disappearance rate was observed was 0.8 μmol/kg. The apparent affinity (Km) of the hepatic transport was higher in Gilbert's syndrome patients than in control subjects (3.61 ± 0.37 μmol sulfobromophthalein/kg vs. 2.76 ± 0.29 μmol sulfobromophthalein/kg, mean ± S.D.; p<0.01), whereas no difference was found in Vmax(0.95 ± 0.11 μmol sulfobromophthalein/kg vs. 0.93 ± 0.10 μmol sulfobromophthalein/kg/min, mean ± S.D.; N.S.). We conclude that a defective sulfobromophthalein hepatic transport is present in Gilbert's syndrome, and it may be revealed by lowering the does of the dye. The data sugest that the defect in Gilbert's syndrome is an impaired affinity for sulfobromophthalein of one or more of the putative transport proteins in the basolateral plasma membrane of the hepatocy

ISSN:0270-9139    

DOI:10.1002/hep.1840120206

出版商:W.B. Saunders    

年代:1990

数据来源: WILEY

摘要:

AbstractChronic liver disease affects up to 20% of children with α‐antitrypsin deficiency owing to the PiZZ genotype. Previous observations of a familial occurrence and abnormal immune responses to liver antigens in these patients suggests that immunoregulatory genes may be involved in the pathogenesis of liver damage. We have identified HLA phenotypes and class II (HLA‐DR) gene polymorphisms in 140 white PiZZ subjects, of whom 92 (83 index patients) had liver disease, and 206 first‐degree relatives. DR3* was present in 35 of 75 (46.7%) unrelated patients with liver disease compared with 5 of 28 (17.8%) patients without (p<0.01) and 23 of 100 controls (p<0.001). DR4 was increased in patients without liver disease; it was present in 17 of 28 (60.7%) compared with 29 of 75 (38.7%) patients with liver disease (p<0.05) and 36 of 100 controls (p<0.025). Using Southern blot analysis with HLA‐DRB and DQB DNA probes, we identified two polymorphisms of DR3, only one (Dw25) of which is raised in PiZZ individuals with liver disease (9 of 55: 16.4%) compared with 1 of 23 (4.4%) without and 2 of 52 (3.9%) controls (p<0.05). Analysis of the segregation of HLA haplotypes in 77 families revealed no concordance for liver disease with HLA in those with affected sibships, indicating that, although DR3‐Dw25 is associated with liver disease in α1‐antitrypsin deficiency, other factors must play a pathogenic role. (HEPATOLOGY199

ISSN:0270-9139    

DOI:10.1002/hep.1840120207

出版商:W.B. Saunders    

年代:1990

数据来源: WILEY

摘要:

AbstractControversy exists regarding major histocompatibility complex antigen expression on hepatocytes. In this study, hepatocyte expression of class I and II major histocompatibility complex antigens was investigated in diseased and normal livers, using indirect immunofluorescent staining of mechanically isolated, viable hepatocytes. Hepatocytes were obtained from 76 children: 10 with autoimmune chronic active hepatitis, nine with primary sclerosing cholangitis, nine with chronic hepatitis B virus infection, five after liver transplantation, 19 with extrahepatic biliary atresia, 11 with α1‐antitrypsin deficiency, four with idiopathic neonatal hepatitis and nine with histologically normal liver. Immunohistochemistry was performed in all cases; flow cytofluorimetry was performed for class I antigens in 38 cases and performed for class II antigens in 18 cases. From three children with autoimmune chronic active hepatitis and two with chronic hepatitis B virus infection, isolated hepatocytes were also incubated with γ‐interferon before staining and analysis. By fluorescence microscopy, class I antigens were detected on hepatocytes from all children, the highest percentage (100%) of positive cells and the most intense staining were observed in untreated patients with autoimmune chronic active hepatitis or primary sclerosing cholangitis and in those with acute rejection of a liver transplant. Reduced class I antigen expression occurred in chronic hepatitis B virus infection. Class II antigens were only detected on hepatocytes from eight patients: three with autoimmune chronic active hepatitis and five with primary sclerosingcholangitis, all untreated. Flow cytofluorimetric analysis confirmed the results obtained by fluorescence microscopy, but it also demonstrated a weak class II antigen expression during liver allograft rejection. After culture with γ‐interferon, the hepatocytes from all five children showed an increase in staining intensity for class I and the acquisition of staining for class II antigens. This study shows that, whereas class I antigens are invariably expressed on liver cells, class II antigens are only found on hepatocytes from untreated patients with immune‐mediated liver disorders in whom class I antigen display is also enhanced. (HEPATOLOGY1990;1

ISSN:0270-9139    

DOI:10.1002/hep.1840120208

出版商:W.B. Saunders    

年代:1990

数据来源: WILEY

摘要:

AbstractCollagen type I and procollagen type III were localized at the ultrastructural level on ultrathin frozen sections of rat liver by the protein A–gold technique using affinity‐purified primary antibodies. Both collagen type I and procollagen type III were localized on nearly all solitary and bundled fibrils in the space of Disse. Simultaneous localization of collagen type I and procollagen type III by a double‐labeling procedure using protein A–gold probes of different sizes unequivocally demonstrated the presence of both collagens in the same fibrils.Measurement of the diameter of large numbers of collagen fibrils in the space of Disse of the rat liver showed a unimodal distribution of the fibril diameters around an average value of 62.4 nm (S.D. = 12.8 nm), and 91% of the collagen bundles contained less than 30 fibrils. Additional measurements on epoxy resin–embedded material of five biopsy specimens of normal human liver showed a comparable unimodal distribution of the fibril diameters around an average value of 57.2 nm (S.D. = 9.6 nm), and 74% of the bundles contained less than 60 fibrils. The latter observation demonstrates that human liver contains broader interstitial collagen bundles than rat liver.From these results, we conclude that the space of Disse of normal rat and human liver contains a uniform population of striated interstitial collagen fibrils. In the rat liver, these fibrils contain both collagen type I and procollagen type III. Therefore the concept that procollagen type III is predominantly localized in small diameter fibrils or bundles, whereas collagen type I is preferentially localized in thick ones, does not hold. (HEPATOLOGY1990;12

ISSN:0270-9139    

DOI:10.1002/hep.1840120209

出版商:W.B. Saunders    

年代:1990

数据来源: WILEY

摘要:

AbstractAdenosine administration was tested in rats with carbon tetrachloride–induced hepatic fibrosis and was able to partially prevent the enlargement of liver and spleen induced by the toxin. This amelioration of the hepatomegaly was accompanied by a 50% reduction of the liver collagen deposition and preservation of content of glycosaminoglycans. A stimulated hepatic collagenase activity is apparently the mechanism for reduction of collagen accumulation. These effects were associated with a striking improvement in liver function. Adenosine treatment did not modify the late hepatotoxic effect of the carbon tetrachloride; however, the stimulatory effect of the nucleoside on energy state appeared to counteract the drastic decreases in adenine nucleotides, ATP, ATP/ADP ratio and energy charge elicited by the hepatotoxin. Moreover, a possible beneficial action of enhanced hepatic oxygenation caused by the vasodilator properties of adenosine cannot be ruled out. Regardless of the mechanism, adenosine seems to change the cellular response to the injury induced by the hepatotoxin. (HEPATOLOGY1990;12:242–2

ISSN:0270-9139    

DOI:10.1002/hep.1840120210

出版商:W.B. Saunders    

年代:1990

数据来源: WILEY

摘要:

AbstractThe distribution of several extracellular matrix components in the liver of patients with acute viral hepatitis was studied by light and electron microscopy using indirect immunoperoxidase methods.Light microscopy revealed type III and type V collagen and fibronectin in the portal tracts and the area of focal necrosis, showing cell infiltration. Type III and type V collagen were more strongly stained in the periphery of focal necrosis. Type IV collagen was seen around the vessels and hepatocytes near the focal necrosis. Electron microscopy showed many transitional Ito cells in the area of focal necrosis and fibroblasts were observed in the portal tracts, showing collagen fiber deposition. Numerous collagen fibrils were observed around fibroblasts, Ito cells and hepatocytes. Using immunoelectron microscopy, type III and type IV collagen and fibronectin were observed in the rough endoplasmic reticulum of Ito cells and hepatocytes localized near the area of focal necrosis or fiber deposition. In addition, type IV collagen was seen in the rough endoplasmic reticulum of endothelial cells forming capillary vessels.These results suggest that several extracellular matrix components such as types III, IV and V collagen and fibronectin, produced by Ito cells, hepatocytes or endothelial cells, play important roles in the healing of liver damage in acute viral hepatitis. (HEPATOLOGY1990; 12:249–256

ISSN:0270-9139    

DOI:10.1002/hep.1840120211

出版商:W.B. Saunders    

年代:1990

数据来源: WILEY