K. Gmelin

Synthesis of the X-protein of hepatitis B virus in vitro and detection of anti-X antibodies in human sera.

A protein of 154 amino acids, predicted to be encoded by the X-open reading frame of the hepatitis B virus (HBV) genome, was synthesized in an in vitro translation system from SP6 transcripts containing the X-coding sequence. As characterized by SDS-PAGE and immunoprecipitation this X-protein possesses the expected molecular weight of 17 kDa and reacts specifically with rabbit antisera directed against a fusion protein from Escherichia coli that contained 145 of the 154 amino acids from the X-sequence. The X-protein, radiolabeled with [35S]methionine, provided a sensitive and specific antigen to screen for anti-X antibodies in sera from HBV patients. Positive signals were obtained preferentially in subjects suffering from HBV-induced liver cirrhosis or primary hepatocellular carcinoma (PHC), i.e., individuals that had been exposed for an extended time period to HBV gene products. Carefully controlled experiments failed to reveal the presence of X-related proteins specific to liver specimens from HBV patients.

Epidemiological studies on the prevalence of hepatitis delta virus infections in the Federal Republic of Germany

This study evaluated the prevalence of hepatitis Delta virus (HDV) infections in various groups of HBsAg carriers including drug addicts and patients with hemophilia in the Federal Republic of Germany. HDV was found only occasionally (less than 1%) in individuals found HBsAg positive during an examination as potential blood donors or in hemodialysis patients, but in 3% in patients with chronic hepatitis and up to 50% in drug addicts and hemophilia patients. These findings are in agreement with data reported from other European countries. Presence of antibodies to HDV in two hemodialysis patients indicates the presence of HDV in this group and screening for HDV infections in hemodialysis units is indicated to prevent outbreaks of this disease in HBsAg-positive patients with possibly serious consequences.

Detection of antibodies against pre-S1 proteins in sera of patients with hepatitis B virus (HBV) infection

Pre-S1 (large S) proteins are components of the envelope of HBV. The presence of pre-S1 proteins is correlated with viral replication. To test sera of patients with HBV infection for the presence of antibodies against pre-S1 proteins (anti-pre-S1), an E. coli extract containing a pre-S fusion protein covering the greater part of the pre-S1 region was subjected to Western blotting and probed with sera of patients. Anti-pre-S1 was present in the sera of all 4 patients with acute self-limited HBV infection and in the sera of 3 patients with a fulminant course. The antibodies could not be detected in the sera of 6 patients with acute HBV infection entering chronicity, in the sera from 10 HBsAg carriers or in the sera of 25 patients with chronic liver disease, positive for HBsAg and antibodies to hepatitis Delta virus. In an additional study, anti-pre-S1 could not be found in 11 sera from 12 patients with previous HBV infection, positive for anti-HBs and anti-HBc. Antibodies to pre-S1 proteins appear at the early stage of acute resolving HBV infection and seem to play a role in the elimination of the virus. The antibodies are absent in the sera of patients with acute HBV infection entering a chronic course and in the sera of chronic HBsAg-carriers.

Biliary Secretion Of Bile Acids, Lipids, And Bilirubin By The Transplanted Liver: A Quantitative Study In Patients On Cyclosporine

To investigate biliary secretion in liver-transplanted patients on cyclosporine treatment we used duodenal perfusion with a nonabsorpable marker. After an overnight fast, 4 women and 5 men were studied for 6 hr at least 6 weeks after orthotopic liver transplantation. The data were compared with those obtained in 6 healthy controls. All transplanted patients received immunosuppressive therapy (corticosteroids, azathioprine, and cyclosporine). Biliary secretion rates of healthy controls were: bile acids 1.58±0.67 mmol/hr, phospholipids 0.27±0.11 mmol/hr, cholesterol 0.11±0.01 mmol/hr, and bilirubin 18±0.7 μmol/hr (mean ± SEM). Liver-transplanted patients excreted 2.6010.4 mmol/hr bile acids, 0.56±0.08 mmol/hr phospholipids, 0.18±0.04 mmol/hr cholesterol, and 22.0±4.5 μmol/hr bilirubin. Analysis of individual bile acids revealed that in the bile of liver transplant patients the percentage of cholic acid was elevated, whereas that of deoxycholic acid was reduced as compared with controls. These findings indicate that in the transplanted liver under immunosuppressive therapy with cyclosporine biliary secretion of bile acids, lipids, and bilirubin is not reduced.

Detection of hepatitis B virus core gene products in sera and liver of HBV-infected individuals

The C gene of hepatitis B virus (HBV) codes for at least two different proteins (p 21c and p 17e). To investigate the expression of C-gene-encoded proteins in vivo, serum and liver samples from HBsAg-positive patients as well as serial serum samples from an HBV-transfected chimpanzee were studied. Antibodies directed against bacterially synthesized C-fusion proteins were used in Western blots to test for the presence of p 21c and p 17e. In serial serum samples from the chimpanzee, p 21c and p 17e were detected concomitantly during the acute phase of the infection. When sera of patients with chronic HBV infection were studied, all sera containing p 17e were found to be positive also for p 21c. Sera positive for HBV DNA but negative for HBeAg were only positive for p 21c, indicating that HBeAg/p 17e is not an absolutely reliable marker for infectivity. In liver tissue specimens from 20 patients with HBV-related liver diseases, p 21c was detected in five cases, indicating viral replication. The p 17e antigen, however, was present only in low amounts in three of these five, suggesting that synthesis of p 21c and p 17e is not strictly coupled. C/Pol-gene-encoded fusion proteins were found in the liver tissue of only one patient with cirrhosis, supporting our previous finding that detectable levels of these proteins are expressed rarely.

Follow-up of patients with hepatitis non-A, non-B: incidence and persistence of anti-HCV depend on route of transmission

Of 32 patients with non-A, non-B hepatitis, 10 (31%) were still anti-HCV-positive 12.8 years after the acute phase of the disease. Seven of the patients (21.9%) still had elevated ALT levels, and among these, 5 out of 5 patients who had been subject to parenteral risk were anti-HCV-positive. In contrast, none of the patients who had not been subject to parenteral risks were positive.

Häufigkeit von Delta-Infektionen in Heidelberg

SummaryThe frequency of delta infection was studied in sera of 203 patients with acute hepatitis B, further 461 hepatitis B virus surface antigen-(HBsAg)-positive patients and 117 HBsAg-negative controls by determination of anti-delta by a competitive enzyme immunoassay. Sera have been collected since 1974. None of the sera of acute hepatitis B was anti-delta-positive whereas seven of the HBsAg-positive carriers were anti-delta-positive. Two of the anti-delta-positive patients had chronic hepatitis, four had liver cirrhosis. One of the anti-delta-positive patients with liver cirrhosis died of liver failure. Risk factors included Italian origin and parenteral routes of infection. All sera of 19 relatives of three anti-delta-positive index cases remained anti-delta-negative.