Arnold W

Liver-cell-membrane autoantibody specific for inflammatory liver diseases.

With an immunofluorescence technique using rabbit hepatocytes isolated by a non-enzymatic method an autoantibody directed against liver-cell-membrance was identified. Sera from 361 patients with various liver diseases and 274 patients with primary non-hepatic diseases-many associated with non-organ-specific auto-antibodies-were examined. The antibody (LMA) was found in 27 out of 72 patients with hepatitis-B-surgace antigen (HBsAg)-negative chronic active hepatitis and in 17 out of 28 patients with HBsAg-negative non-alcoholic cirrhosis. Only two patients had LMA and HBsAg, and both had chronic active hepatitis. One patient with extrhepatic disease was found to have LMA, and this patient had biochemical evidence of liver disease. Hence there is a close correlation between the presence of LMA and HBsAg-negative chronic inflammatory liver diseases and its detection may help in diagnosis.

Anti-HBc, HBeAg and DNApolymerase activity in healthy HBsAg carriers and patients with inflammatory liver diseases

ZusammenfassungIn der vorliegenden Arbeit wird über anti-HBc-Titer-Bestimmungen, HBeAg, DNApolymerase-Aktivität im Serum sowie über intrazellulär nachweisbares HBcAg bei gesunden HBsAg-Trägern und Patienten mit HBsAg-positiven entzündlichen Lebererkrankungen berichtet.32/44 Patienten mit akuter Virus-B-Hepatitis hatten keine anti-HBc-Titer-Erhöhung in der ersten Krankheitswoche. Bei gesunden HBsAg-Trägern schwanken die anti-HBc-Titer zwischen 1:10 und 1:32000 (mittlerer Titer 1:4000). Bei HBsAg-positiven chronisch-aktiven Hepatitiden fand sich ein mittlerer Titer zwischen 1:32000 bis 1:64000, bei HBsAg-positiver CPH ein mittlerer Titer von 1:16000. Alle Patienten mit autoimmuner CAH (HBsAg-negativ) hatten anti-HBc-Titer unter 1:10.In einer Gruppe von 71 asymptomatischen HBsAg-Trägern konnten immunhistologisch bei keinem der gesunden HBsAG-Träger HBcAg im Leberzellkern nachgewiesen werden. Im Gegensatz dazu wurde HBcAg bei 4/5 HBsAg-positiven CAH- und 6/9 CPH-Patienten gefunden.Eine Erhöhung der DNApolymerase-Aktivität wurde bei keinem der gesunden HBsAg-Träger nachgewiesen. Von den 44 HBsAg-positiven akuten Virus-B-Hepatitiden hatten nur 3 Fälle eine Erhöhung der DNApolymerase-Aktivität. Andererseits fand sich eine DNApolymerase-Aktivität bei 17/37 Patienten mit HBsAg-positiver CAH und 9/15 Fällen mit CPH.Die Untersuchungen konnten eine enge Korrelation zwischen dem Nachweis des HBeAg im Serum und der DNApolymerase-Aktivität zeigen.Die Befundmuster ermöglichen eine klare Abgrenzung der “gesunden” HBsAg-Träger von den HBsAg-Trägern mit entzündlichen Lebererkrankungen.SummaryIn this paper we report on anti-HBc-titers, HBeAg, DNApolymerase activity in the serum and intracellular HBsAg in healthy HBsAg-carriers and patients with HBsAg-positive inflammatory liver diseases.32/44 patients with acure virus-B-hepatitis were negative for anti-HBc in the first week of the disease. Anti-HBc-titers in healthy HBsAg-carriers varied between 1:10 and 1:32,000 (medium titer 1:4,000). In HBsAg-positive CAH we found a medium titer between 1:32,000 and 1:64,000, in cases with CPH of about 1:16,000. All autoimmune type CAH showed anti-HBc-titers less than 1:10.By immunofluorescence we could demonstrate in a group of 71 asymptomatic HBsAg-carriers in none of the healthy HBsAg-carriers HBcAg in the liver cell nuclei. In contrast HBcAg could only be found in 4/5 HBsAg positive CAH- and 6/9 CPH patients.No elevated DNApolymerase activity could be demonstrated in healthy HBsAg-carriers. Out of 44 patients with virus-B-hepatitis only 3 showed elevated DNApolymerase activity. On the other hand DNApolymerase elevation was demonstrable in 17/37 cases with CAH and 9/15 with CPH.The investigations showed a strong correlation between the demonstration of HBeAg in the serum and the DNApolymerase activity.The characteristic findungs enabled us to differentiate between “healthy” HBsAg-carriers and HBsAg-carriers with inflammatory liver diseases.

Localisation of e-antigen in nuclei of hepatocytes in HBsAg-positive liver diseases.

By the direct immunofluorescence technique 19 liver biopsies were examined for the presence of the e-antigen associated with the hepatitis B virus infection. With the use of double incubation and blocking experiments with FITC and Rhodamine labelled antisera against hepatitis B core antigen and e-antigen, evidence is presented that the e-antigen is localised in the nuclei of the hepatocytes. We conclude that the e-antigen and the core antigen are not identical, although they are often present simultaneously in the nucleus of the hepatocyte.

Inhibition of hepatitis-B-virus DNA polymerase by phosphonoformate: Studies on its mode of action

Phosphonoformate (PFA) and phosphonoacetate (PAA) were tested for their ability to inhibit the hepatitis‐B‐virus associated DNA polymerase. The HBV DNA polymerase was inhibited by 100 μM/liter PFA 50% while it was highly resistant to PAA. The inhibition of the Dane particle‐associated DNA polymerase by PFA was not competitive to substrates and not affected by changes in the magnesium concentration. PFA was active also after initation of the DNA polymerase reaction. Competition studies revealed that PFA had a higher affinity to a proposed pyrophosphate binding site than PAA or—alternatively—that both compounds bind to different sites.

IgM antibody to hepatitis B core antigen (anti-HBc IgM) in “healthy” HBsAg carriers: A longitudinal study of 75 cases

SummaryIn 75 healthy HBsAg carriers with normal liver tissue who were followed over a four years period, anti-HBc IgM was determined by ELISA. 61 HBsAg carriers (81%) were positive for anti-HBc IgM at first investigation. 54 individuals demonstrated persistence of anti-HBc IgM, 7 became anti-HBc IgM-negative within the observation period. 12 persons were persistent anti-HBc IgM-negative, and 2 developed anti-HBc IgM of low quantities. 3 of 4 individuals with HBsAg clearance demonstrated a considerable decrease of anti-HBc IgM concentration. Although signs of liver damage or development of chronic liver diseases were not observed at the time of control biopsy the existence of anti-HBcIgM indicates that there exists also in healthy HBsAg carriers a persistent stimulation by HBcAg, inducing the production of anti-HBc IgM as a sign of permanent ongoing virus B replication. These results require a new and critical review of the “healthy” HBsAg carrier state.ZusammenfassungBei 75 gesunden HBsAg-Trägern mit bioptisch normalem Lebergewebe, die über einen Zeitraum von 4 Jahren beobachtet wurden, wurde anti-HBc IgM mit der ELISA-Technik bestimmt. Anti-HBc IgM fand sich bei der Erstuntersuchung bei 61 HBsAg-Trägern (81%). 54 Personen blieben anti-HBc IgM positiv, 7 wurden negativ. Bei 12 HBsAg-Trägern war von Anfang an kein anti-HBc IgM nachweisbar, und bei 2 Personen fanden sich erst bei der Kontrolluntersuchung geringe Mengen von anti-HBcIgM. 3 von 4 HBsAg-Trägern, die innerhalb des Beobachtungszeitraumes HBsAg aus dem Serum eliminerten, zeigten einen deutlichen Abfall der anti-HBc IgM-Konzentration oder wurden anti-HBc IgM negativ. Obwohl eine Verschlechterung des morphologischen Leberbefundes oder die Entwicklung einer chronischen Leberentzündung bei der Kontrollbiopsie nicht beobachtet wurde, muß aufgrund des positiven Nachweises von anti-HBc IgM bei gesunden HBsAg-Trägern angenommen werden, daß auch bei diesen Personen eine ständige aktive Virus B Replikation stattfindet. Diese Ergebnisse erfordern eine kritische Neubewertung des “gesunden” HBsAg-Trägerstatus.

The clinical relevance of the antibody to hepatitis B core antigen (anti-HBc): A review

The antibody against the core component of the Dane particle (anti-HBc) is generally detected in the sera of individuals with acute type B hepatitis and in chronic HBsAg carriers. While the serological demonstration of HbsAg with or without anti-HBc indicates continued replication of viral antigens, the co-occurrence of anti-HBs and anti-HBc is considered a marker of recent HBV replication. The demonstration of anti-HBc in the absence of HBsAg and anti-HBs is in agreement with at least four different states of HBV infection. As this pattern indicates persistent HBV infection in some cases and recovery from an acute type B hepatitis in others, current efforts focus on further characterization of this pattern, using additional test methods such as anti-HBe and anti-HBc of the IgM class.

Comparison of indicators for dane particles.

ZusammenfassungDas Dane Partikel wird als das komplette Hepatitis B Virus angesehen. Seren mit einer großen Zahl von Dane Partikeln gelten als hoch infektiös. In der vorliegenden Studie haben wir die Höhe der DNA Polymerase Aktivität — gemessen in 20fach Dane Partikel Konzentraten — mit dem HBsAg, dem HBeAg und dem anti-HBc Titer verglichen. Unsere Untersuchungen zeigen, daß der HBeAg Titer ein semiquantitatives Maß für die Zahl zirkulierender Dane Partikel darstellt. Die HBsAg Konzentration steigt ebenfalls mit der Zahl im Serum zirkulierender Dane Partikel an und stellt ebenso — wenn in HBeAg positiven Seren gemessen — ein semiquantitatives Maß für die Zahl zirkulierender Dane Partikel dar. Zwischen dem anti-HBc Titer und dem Serum Dane Partikel Gehalt besteht keine Beziehung.SummaryThe Dane particle is — because of its characteristics — believed to be the complete hepatitis-Bvirus. Sera containing high numbers of Dane particles were shown to be highly infectious. In the present study we related the HBeAg-, the HBsAg- and the anti-HBc titer to the level of DNA polymerase activity measured in 20 fold Dane particle concentrates. The data obtained indicate that the HBeAg concentration gives a semiquantitative estimate on the number of circulating Dane particles. Mean DNA polymerase activity was found to increase with HBsAg concentration and is therefore also of value-if determined in a HBeAg positive serum- for quantitation of Dane particles. The anti-HBc titer was found to be unrelated to the number of circulating Dane particles.

Inhibition of hepatitis B virus specific DNA polymerase by intercalating agents

Intercalating agents, some of them in clinical use, were tested for their ability to inhibit the hepatitis B virus specific DNA polymerase reaction. Ethidium bromide was shown to be the strongest inhibitor among the compounds tested. Compounds in clinical use inhibited the DNA polymerase test only at high concentrations. The inhibitory activity of all compounds tested was increased when the MgCl2 content in the reaction mixture was lowered. UV absorption studies presented no evidence that this effect was due to complex formation of magnesium and the individual compounds. The therapeutic significance of these findings is not certain and needs further work.

Etiology of Hepatitis B Surface Antigen (HBsAg)-Negative Chronic Hepatitis

A study was undertaken to elucidate the etiology of HBsAg-negative chronic hepatitis. Form 37 individuals with HBsAg-negative chronic hepatitis, 11 had liver membrane autoantibody (LMA) and were thus classified as autoimmune. 6 patients had anti-HBc, 1 of which was also positive for LMA. The majority of individuals with HBsAg-negative chronic hepatitis had antibodies to hepatitis A antigen (anti-HAV), in general at low titer. We conclude from our data that hepatitis A and hepatitis B virus infections are unlikely to play a significant role in inducing or maintaining HBs-Ag-negative chronic hepatitis. The etiological role of non-A non-B hepatitis agent(s) is difficult to estimate and must await the detection of appropriate markers for type non-A non-B hepatitis.

Quantitation of HBeAG and anti-HBe by RIA in sera of chronic HBsAG carriers and individuals with type B hepatitis.

SummaryThis paper gives the results for HBeAg and anti-HBe titers in chronic HBsAg carriers and patients with type B hepatitis using a “solid-phase” radioimmunoassay.In tumor and hemodialysis patients the HBeAg titers are statistically significant higher compared to the group of HBsAg positive CAH or CPH.High anti-HBe titers are a characteristic finding in “healthy” HBsAg carriers. On the other hand, there is a subgroup of HBsAg positive CAH with anti-HBe; although there are signs of an ongoing virus B replication these cases of CAH proceed sometimes to cirrhosis.ZusammenfassungIn dieser Arbeit werden die Ergebnisse der HBeAg und anti-HBe-Titer Bestimmungen mit einem „solid-phase“-Radioimmunoassay dargestellt.Bei Tumor- und Hämodialysepatienten sind die HBeAg-Titer statistisch signifikant höher als bei Patienten mit HBsAg positiver CAH oder CPH.Hohe anti-HBe Titer sind ein charakteristischer Befund für „gesunde“ HBsAg-Träger. Andererseits gibt es eine Untergruppe der HBsAg positiven CAH mit anti-HBe; obgleich bei dieser Form eine CAH keine Zeichen einer fortbestehenden Virus B-Replikation nachgewiesen werden kann, schreitet die Krankheit manchmal in Richtung auf eine Zirrhose fort.