摘要:

AbstractWe have previously identified four of the M2 antigens in primary biliary cirrhosis as the E2 components (dihydrolipoamide acyltransferases) of pyruvate dehy‐drogenase complex, branched‐chain 2‐oxo acid dehy‐drogenase complex and 2‐oxoglutarate dehydrogenase complex and the protein X component of pyruvate dehydrogenase complex (approximate molecular masses: 74, 50, 50 and 52 kD, respectively). In the present study, we have examined by immunoblotting the frequency of IgG and IgM autoantibodies to these four proteins in 129 patients with primary biliary cirrhosis (36 histological Stage I, 42 Stage II/III, 51 Stage IV) and 77 controls (49 non–primary biliary cirrhosis chronic liver disease, 16 primary Sjögren's syndrome, 12 healthy normal women). One hundred twenty‐seven of 129 (98%) primary biliary cirrhosis patients had antibodies against at least one of the four M2 polypeptides, compared to 2/77 controls (both had autoimmune chronic active hepatitis and were antimitochondrial antibody positive by indirect immunofluorescence). One hundred twenty‐one of 129 (94%) primary biliary cirrhosis sera reacted with the E2 component and protein X of pyruvate dehydrogenase complex, 69/129 (53%) primary biliary cirrhosis sera reacted with E2 of branched‐chain 2‐oxo acid dehydrogenase complex and 113/129 (88%) reacted with E2 of 2‐oxoglutarate dehydrogenase complex. Primary biliary cirrhosis patients with histological Stage I disease had a lower incidence of autoantibodies to each M2 protein, compared to more advanced disease (IgG, p<0.05) but only 2/36 Stage I patients had no anti‐M2 antibodies. There was no correlation between the presence of IgG or IgM antibodies to the M2 polypeptides and established prognostic markers in primary biliary cirrhosis (serum bilirubin and albumin levels). The specificity of autoantibodies to the E2 components of all three 2‐oxo acid dehydrogenase complexes for primary biliary cirrhosis suggests that a common mechanism or immunological defect underlies the development

ISSN:0270-9139    

DOI:10.1002/hep.1840100402

出版商:W.B. Saunders    

年代:1989

数据来源: WILEY

摘要:

AbstractTo determine whether abnormalities of lymphocyte function in primary biliary cirrhosis are due to altered function of immunoregulatory T cell subpopulations, phenotypic and functional characteristics of CD4+ T cells were examined. The proportion of CD4+ T cells expressing the Leu‐8 and CD45R antigens was normal in patients with primary biliary cirrhosis. The capacity of CD4+, Leu‐8‐T cells to provide helper function for pokeweed mitogen‐stimulated immunoglobulin synthesis by B cellsin vitrowas similar in patients and controls. However, in contrast to normal individuals and patients with other liver diseases, CD4+, Leu‐8+ T cells from six of 10 patients with primary biliary cirrhosis did not suppress, but enhanced immunoglobulin synthesis. Whereas treatment of CD4+ T cells from normal individuals with anti‐Leu‐8 monoclonal antibody enhanced their suppressor function, similar treatment of CD4+ T cells from patients with primary biliary cirrhosis did not increase their suppressor function. To determine whether the abnormal regulatory function of CD4+, Leu‐8+ T cells was due to a defect of cell activation, the proliferative response of CD4+ T cell subpopulations to mitogenic stimulation was examined. The proliferative responses of CD4+, Leu‐8‐T cells from patients with primary biliary cirrhosis and controls were similar, but the proliferative responses of CD4+, Leu‐8+ T cells from patients with primary biliary cirrhosis were lower than those of control cells. Since the CD4+, Leu‐8+ T cell population plays a role in suppressing immunoglobulin synthesis and is contained within the autoreactive T cell population, the abnormal function of this T cell subpopulation in some patients with primary biliary cirrhosis may play a role in defective immunoregulatio

ISSN:0270-9139    

DOI:10.1002/hep.1840100403

出版商:W.B. Saunders    

年代:1989

数据来源: WILEY

摘要:

AbstractWe have compared the effect of ursodeoxycholic acid with placebo on the clinical state, blood liver chemistries and serum and urinary bile acids in four patients with primary biliary cirrhosis. All parameters were evaluated monthly, and bile acid composition was measured by capillary gas‐liquid chromatography. At the time of admission, all patients showed intense pruritus, and their serum alkaline phosphatase, AST and ALT levels were elevated 4.3, 2.7 and 2.3 times over control values. Serum bile acids were elevated almost 38‐fold with 2.5 times more cholic acid than chenodeoxycholic acid. Urinary bile acid output was elevated 28 times the control values, and 36% were 1β‐hydroxycholic acid, 1β‐hydroxydeoxycholic acid and hyocholic acid (3α,6α,7α‐trihydroxy‐5β‐cholanoic acid). Three months of placebo administration did not significantly affect the clinical or biochemical presentations, and the serum and urinary bile acid composition did not change. In contrast, ursodeoxycholic acid feeding (12 to 15 mg per kg per day) for 6 months abolished pruritus in two and lessened itching in two subjects and reduced serum alkaline phosphatase, AST and ALT levels by 21, 35 and 47%, respectively. The mean values for the total serum bile acid concentrations in these patients declined 26% from the pretreatment value, but the proportion of ursodeoxycholic acid increased from 3 to 40% of the total bile acids; thus, total fasting serum endogenous bile acid levels decreased almost 50%. Similar changes were noted in the urinary bile acids, in which ursodeoxycholic acid became the major bile acid, and approximately 18% were hydroxylated at C‐1, C‐6 and C‐21. These results demonstrate that, compared with placebo, ursodeoxycholic acid significantly lowers serum and urinary bile acids in primary biliary cirrhosis and produces considerable clinical an

ISSN:0270-9139    

DOI:10.1002/hep.1840100404

出版商:W.B. Saunders    

年代:1989

数据来源: WILEY

摘要:

AbstractA randomized, double‐blind, 1‐year pilot study of prednisolone treatment for primary biliary cirrhosis was undertaken. Nineteen patients received 30 mg prednisolone per day initially, with a maintenance dose of 10 mg per day. Seventeen patients received placebo. The groups were matched for age, menopausal status, hepatic histological stage and bilirubin.Treatment was well tolerated without dropouts. Two patients receiving prednisolone developed diabetes, one a duodenal ulcer and one depression. One patient receiving placebo died of liver failure after 3 months.Cholestatic symptoms (itch and fatigue) improved on prednisolone. There was significant (prednisolone vs. placebo) improvement in transaminase (p = 0.0214), alkaline phosphatase (p = 0.0032), procollagen III peptide (p = 0.0103), immunoglobulin G (p = 0.0012) and liver histology (p = 0.016); these changes were greatest among noncirrhotic patients.No patient developed skeletal symptoms. Fifty‐seven per cent had abnormal triolein breath tests prior to treatment, and 65% had abnormally low calcium absorption tests. Calcium absorption increased significantly in the treated group vs. placebo at 2 weeks (p<0.02), but not at 1 year. Femoral photon absorptiometry fell in the prednisolone group after 1 year (‐3.5% vs. placebo +0.5%, p<0.05), as did trabecular bone volume (‐6% vs. ‐2.8%, p<0.005) and resorption surface (‐11% vs. +2%, p<0.02) on serial bone biopsy.Prednisolone seems to exert a favorable hepatic effect in primary biliary cirrhosis but at the expense of increased bone loss to approximately twice the expected rate. Prednisolone treatment merits further assessment in primary biliary cirrhosis over a longer period, with attention to selection of patients most likely to benefit and continuing observation of bone mass to better establish the “cost

ISSN:0270-9139    

DOI:10.1002/hep.1840100405

出版商:W.B. Saunders    

年代:1989

数据来源: WILEY

摘要:

AbstractThe natural history of primary sclerosing cholangitis was assessed in 174 patients; 37 were asymptomatic and 137 had symptoms related to underlying liver disease. At the time of diagnosis, the mean age was 39.9 years, 66% of the primary sclerosing cholangitis patients were male and 71% had associated inflammatory bowel disease, most commonly chronic ulcerative colitis. Longterm follow‐up (mean: 6.0 years; range: 2.7 to 15.5 years) was available in all patients. During follow‐up, 59 (34%) of the patients died: 55 in the symptomatic group and four in the asymptomatic group. Median survival from the time of diagnosis of primary sclerosing cholangitis at the Mayo Clinic was 11.9 years. Survival in the asymptomatic group was significantly decreased compared with that in a control population matched for age, race and sex. Multivariate analysis (Cox proportional hazards regression modeling) revealed that age, serum bilirubin concentration, blood hemoglobin concentration, presence or absence of inflammatory bowel disease and histologic stage on liver biopsy were independent predictors of high risk of dying.The development of a multivariate statistical survival model is a major step in identifying individual primary sclerosing cholangitis patients at low, moderate and high risk of dying. Such models will be useful for stratifying patients in therapeutic trials, in patient counseling and in patient selection and timing of liver transplantat

ISSN:0270-9139    

DOI:10.1002/hep.1840100406

出版商:W.B. Saunders    

年代:1989

数据来源: WILEY

摘要:

AbstractPerivenular hepatocytes are the first cells within the liver lobule to display signs of toxicity following long‐term alcohol use. In an attempt to explain this phenomenon, we have examined the hepatic intralobular distribution in rats and man of P450IIE1, a P‐450 isozyme that not only oxidizes ethanol but is also inducible by this agent. Frozen liver sections and microsomes were prepared from male Sprague‐Dawley rats pair‐fed liquid diets containing 36% of total calories as either ethanol or carbohydrate (control) for 10 to 21 days. Frozen sections or microsomes were also prepared from liver biopsy samples obtained from 17 male patients with diverse drinking histories. Immunohistochemical staining was performed using the peroxidase‐antiper‐oxidase method after liver sections were reacted with monospecific antibody (IgG) directed against human P450IIE1. Immunoreaction intensity was blindly rated in order to provide a semiquantitative assessment of P450IIE1 levels in perivenular, midzonal and periportal hepatocytes. At low applied anti‐P450IIE1 IgG concentrations (2.5 μg per ml), P450IIE1 immunostaining was observed exclusively within the perivenular area in sections from all ethanol‐treated rats, whereas no visible immunoreaction was found in sections from their pairfed controls. At higher applied antibody concentrations (15 μg per ml), panlobular antigen immunostaining was observed in five of the six ethanol‐treated animals, and P450IIE1 could now also be detected in perivenular hepatocytes from the control rats. In accordance with these immunohistochemical findings, protein blotting with anti‐P450IIE1 IgG revealed a 7.5‐fold increase in liver microsomal P450IIE1 content in ethanol‐treated animals when compared to their pair‐fed controls. With human liver, perivenular P450IIE1 immunostaining was observed only in biopsy sections obtained from recently drinking alcoholics (abstinence period of 1 day) when limiting concentrations (5 μg per ml) of the primary antibody were used. Increasing the applied anti‐P450IIE1 IgG concentration to 15 μg per ml resulted in perivenular staining of the immunogen in liver sections from abstinent alcoholics (abstinence period of 4 to 8 days) and nondrinkers as well. Immunoblot analysis of human liver microsomes disclosed that the hepatic microsomal P450IIE1 content in recently drinking alcoholics was 4‐fold higher than that found in nondrinkers. Our results show that, in both rats and in man, P450IIE1 is normally localized within the perivenular region, or zone 3, of the liver lobule, and that induction of P‐450IIE1 by prolonged alcohol consumption occurs primarily within the same acinar region. Since P450IIE1‐catalyzed ethanol oxidation results in the formation of acetaldehyde, an established cytotoxin, and free radicals capable of peroxidizing cell membranes, the elevated P450IIE1 levels found in perivenular hepatocytes after chronic alcohol intake suggest that increased production of ethanol‐derived toxic metabolites occurs in zone 3 cells, which, in turn, may explain their enhanced su

ISSN:0270-9139    

DOI:10.1002/hep.1840100407

出版商:W.B. Saunders    

年代:1989

数据来源: WILEY

摘要:

AbstractTransport of taurocholate into the hepatocyte against unfavorable chemical and electrical gradients occurs via a sodium‐dependent, carrier‐mediated transport system. Although this cotransporter has been characterized in the rodent, it has not been demonstrated in man. Therefore, we utilized human liver, obtained via multiorgan donation but not used for transplantation, to prepare basolateral (sinusoidal) liver plasma membrane vesicles by a Percoll gradient method. Na+,K+‐ATPase, a marker enzyme for the basolateral domain, was enriched 28.9‐fold in the final membrane fraction compared with homogenate, whereas the bile canalicular membrane enzymes Mg++‐ATPase and alkaline phosphatase were enriched only 3.4‐ and 6.4‐fold, respectively. Marker enzyme activities for endoplasmic reticulum, lysosomes and mitochondria were not enriched compared with homogenate. Integrity of the membrane vesicles was confirmed by the demonstration of Na+‐dependent concentrative uptake of the amino acid L‐alanine (estimated intravesicular volume of 0.59 μl per mg protein). An inwardly directed 100 mMNa+gradient stimulated the initial rate of 2.5 μMtaurocholate uptake and energized a transient 2‐fold accumulation of the bile acid above equilibrium (“overshoot”). In contrast, uptake was slower and no overshoot occurred with a K+gradient. A negative intravesicular potential, created by altering accompanying anions or by valinomycin‐induced K+diffusion potentials, did not enhance taurocholate uptake, suggesting an electroneutral cotransport mechanism. Chloride as the accompanying anion stimulated the initial rate of uptake compared with anions of lesser or greater lipid permeability. Na+‐dependent taurocholate (4 μM) uptake was significantly inhibited by 250 μMcholate, taurocholate, glycocholate, taurochenodeoxycholate and bromsulfophthalein. Conversely, in the presence of a K+gradient, only taurochenodeoxycholate and bromsulfophthalein significantly inhibited taurocholate uptake. The initial rate (5 sec) of Na+‐dependent taurocholate uptake, measured as a function of extravesicular taurocholate concentration (1 to 200 μM), demonstrated Michaelis‐Menten kinetics with a Kmof 33.8 ± 4.4 μMand a Vmaxof 1.25 ± 0.99 nmoles per mg protein per min. We conclude from these studies that taurocholate uptake by basolateral plasma membrane vesicles from human liver is sodium dependent, electroneutral, inhibitable by other bile acids and saturable, with an apparent Kmnear the concentration of t

ISSN:0270-9139    

DOI:10.1002/hep.1840100408

出版商:W.B. Saunders    

年代:1989

数据来源: WILEY

摘要:

AbstractEndotoxins play an important role in the pathogenesis of complications of surgery in obstructive jaundice. Preoperative treatment with orally administered deoxycholic acid prevented endotoxin‐related complications, such as renal malfunction. Other bile acids, however, were less effective, and the mechanism of action is not known.Endotoxin toxicity is considered to be largely mediated by tumor necrosis factor/cachectin, a cytokine release by mononucler phagocytes. Therefore, we studied the influence of different bile acids on endotoxin‐induced tumor necrosis factor production by monocytesin vitro.Bile acids inhibit tumor necrosis factor production through a direct inhibitory effect on the monocytes. Deoxycholic acid was the most effective, chenodeoxy‐cholic acid was less effective and ursodeoxycholic acid was ineffective in the concentrations used. Bile acids did not inactivate endotoxin as measured in a chromogenicLimulusamebocyte lysate assay.The therapeutic effect of bile acids in obstructive jaundice can be explained by an inhibition of endotoxin‐induced tumor necrosis factor release by mononuclear pha

ISSN:0270-9139    

DOI:10.1002/hep.1840100409

出版商:W.B. Saunders    

年代:1989

数据来源: WILEY

摘要:

AbstractPrimary duck hepatocytes obtained from Pekin ducks congenitally infected with duck hepatitis B virus were used to monitor expression of viral proteins and replication of viral DNA in cell culture. Duck hepatitis B virus core antigen, duck hepatitis B virus pre‐surface antigen and duck hepatitis B virus DNA were detectable for at least 12 days after cell plating. Whereas expression of duck hepatitis B pre‐surface antigen was constant during this time, expression of duck hepatitis B core antigen and of viral DNA rapidly declined. This diminished production of viral components invitrowas paralleled by a change of the hepatocytes toward a fibroblast‐like morphology. Supplementation of cell culture medium with 2% dimethyl sulfoxide, a solvent known to maintain the differentiated state of cultured cells, retained competence of the cultured hepatocytes to express duck hepatitis B core antigen and duck hepatitis B virus DNA at high levels. In a second set of experiments, duck hepatitis B virus negative hepatocytes were infected with duck hepatitis B virus from serum of congenitally infected ducks. Dimethyl sulfoxide remarkably improved the competence of cultured duck hepatocytes to become productively infected. This function was maintained for at least 12 days postpl

ISSN:0270-9139    

DOI:10.1002/hep.1840100410

出版商:W.B. Saunders    

年代:1989

数据来源: WILEY

摘要:

AbstractAn epidemic of viral hepatitis, serologically characterized as due to non‐A, non‐B hepatitis, occurred in a village of South Delhi, India, in December, 1986, through January, 1987. Water contaminated with fecal matter was the apparent source of infection. Disease‐associated virus‐like particles were detected by immune electron microscopy in the feces of three patients within 5 days of illness. The virus‐like particles were agglutinated by autologous acute‐phase serum but not by convalescent serum. Rhesus monkeys inoculated with particle‐containing fecal suspensions developed biochemical and morphologic features of acute, self‐limited hepatitis. The findings in the present study and in earlier investigations of sporadic non‐A, non‐B hepatitis suggest that (i) the epidemic form and a proportion of sporadic cases of this infection in India may be related, both being enterically transmitted and associated with infection by a 27‐to 32‐nm virus‐like particle, (ii) antibody responses to this virus occur early in disease and are transient and (iii) the rhesus monkey may prove to be a suitable model for studies of epidem

ISSN:0270-9139    

DOI:10.1002/hep.1840100411

出版商:W.B. Saunders    

年代:1989

数据来源: WILEY