A randomized controlled trial of a 12-month course of recombinant human interferon-alpha in chronic delta (type D) hepatitis: a multicenter Italian study.

To determine whether long-term therapy with recombinant interferon-alpha can improve the course of chronic delta hepatitis, 61 Italian patients with this disease were randomly assigned to receive either interferon-alpha-2b three times a week (5 MU/m2 for 4 mo and then 3 MU/m2 for another 8 mo) or no treatment. At the end of the 12-mo study, all patients were followed-up for 12 additional months. Normalization or decrease of more than 50% from baseline of serum ALT levels occurred in 42% of treated patients the fourth month of therapy, 26% the twelfth month and 3% the twenty-fourth month vs. 7%, 7% and 0%, respectively, in the control group. However, relapses occurred in 7 of 8 (87.5%) responders 1 to 10 mo (mean = 3.5 mo) after cessation of therapy. Liver biopsies were carried out at baseline and during the twelfth month of treatment. Histological improvement, mostly caused by decrease of portal inflammation, was observed in 57% of treated and 36% of untreated patients. Measures of antiviral activity (serum hepatitis delta virus RNA and intrahepatic hepatitis delta antigen) showed similar levels in treated and control patients. In treated patients the percentage of patients who were negative for HDV RNA never exceeded that of baseline. Although interferon-alpha in the dosage given in this study had no antiviral effect on patients with chronic hepatitis D, it reduced hepatic inflammation as measured by ALT levels. Whether a longer duration or reinstitution of interferon-alpha therapy would achieve long-term control of ALT levels and prevent chronic liver damage is not known.

The absence of hepatitis B virus DNA in hepatitis B e antigen positive sera from chronic hepatitis B surface antigen carriers in China.

Sera from 20 Chinese patients with chronic hepatitis B were examined for hepatitis B e antigen and hepatitis B virus (HBV) DNA. There was considerable discordance with HBV DNA not being detectable in 10 out of 13 (77%) patients who were hepatitis B e antigen positive. Further testing for anti-HBe and HBV-DNA polymerase activity confirmed the results. Possible reasons for this discordance are discussed but neither hepatitis D (delta) infection nor the acquired immunodeficiency syndrome (AIDS) could be implicated.

Combined immunoprophylaxis in the prevention of perinatal transmission of hepatitis B and hepatitis D virus infections in Saudi children.

The efficacy of the combined immunoprophylaxis approach (passive plus active immunization) was evaluated in babies born to 52 HBsAg-carrier Saudi mothers, of whom seven (13.5%) were HBeAg-positive and seven were anti-HDV-positive. Newborns were given 100 IU of hepatitis B immune globulin (HBIG) from the Hepuman Berna-Swiss Serum and Vaccine Institute, Berne, and hepatitis B vaccine (5 micrograms) within 12 hours of birth, and the vaccine was given again at 1 and 6 months of age (5 micrograms/injection). After 18 months of follow-up, all but one baby had protective levels of antibody to HBsAg and none of the babies born to anti-HDV-positive carrier mothers showed evidence of HDV infection. These results show that the combined immunoprophylaxis approach is quite successful in protecting against perinatal transmission of HBV and HDV in the Saudi population. Furthermore, early vaccination against HBV will also protect against HDV infection later in life.

Hepatitis delta virus infection in French male HBsAg-positive homosexuals.

The prevalence and serological features of hepatitis delta virus infections were studied in a French population of chronic, non-drug addict, hepatitis B surface antigen carriers: 42 male homosexual patients were compared to 30 nonhomosexuals (20 who evidently had not been exposed to any of the usual hepatitis B virus-hepatitis delta virus risk factors and 10 hemodialyzed patients or kidney allograft recipients). Six of the 42 male homosexuals (14.3%) had at least one serological marker of hepatitis delta virus infection (hepatitis delta antigen, total and/or IgM anti-hepatitis delta antibodies). Serological follow-up was obtained for five of these patients over several months and confirmed the chronicity of the delta infection in at least four of the five subjects. Hepatitis delta antigen and hepatitis delta virus RNA were found in the liver and in the serum, respectively, of four of the five tested patients. Hepatitis B virus DNA was negative in the serum of five of the six hepatitis delta virus-positive homosexuals vs. only eight of 35 hepatitis delta virus-negative homosexuals (p less than 0.02). Human immunodeficiency virus was negative in all of the nonhomosexuals; its prevalence did not differ between the hepatitis delta virus-positive and -negative homosexuals: three were human immunodeficiency virus-positive among the six former vs. 15 among the 36 latter. Human immunodeficiency virus positivity was without obvious influence on hepatitis B virus replication, since among 35 hepatitis delta virus-negative homosexuals hepatitis B virus DNA was found in 80% of the human immunodeficiency virus-positive individuals and 70% of those who were human immunodeficiency virus-negative.

[Epidemiologic and clinical study of infection caused by hepatitis delta virus in the health service area of La Coruna]. / Estudio epidemiológico y clínico de la infección por el virus de la hepatitis delta en el área sanitaria de La Coruña.

A prospective study on 175 hepatitis HBsAg positive patients was carried out between March 1986 and June 1987 in order to study the epidemiological and clinical aspects of the HVD infection. The global prevalence of this infection was 19.42%, with a statistically relevant predominance amongst intravenous drug addicts (IVDA). Moreover, in most cases it was associated with liver disease (91.17%). Chronic liver disease had a greater incidence amongst HVD positive infected patients and presented more severe clinical and biochemical aspects than other chronic liver diseases without HVD infection. The HVD infection was confirmed in 39.4% of the HBs Ag positive acute hepatitis, being in the majority of the cases delta coinfections with a clinical and biochemical evolution similar to the hepatitis caused by the B virus only. The overinfections evolved to cronicity in all cases. No delta infection occurred amongst immunodepressed patients, and there was an incidence amongst hemophiliacs of 40%. In view of our results, we may conclude that in our community the parenteral route is the main mechanism for the transmission of HVD.

Immunohistochemical research on HDV infection in Chinese patients with chronic liver disease.

The prevalence of HDAg in the liver of Chinese patients with chronic hepatitis and hepatocellular carcinoma was determined by using direct immunofluorescence and immunoperoxidase. Six patients (6.3%) out of 95 HBsAg carriers with inflammatory liver disease and neoplasia were found to be HDAg positive. HDAg was shown in the liver of 6 patients (7.6%) among 79 patients with chronic hepatitis. The relative frequency of HDAg in cirrhosis-B, chronic active hepatitis B and chronic persistent hepatitis B was 14.3%, 7.1% and 5.9% respectively. These results suggest that a sizeable number of HBsAg carriers are superinfected with HDV. In view of the large amount of HBV carriers in China, the relative minor but distinct presence of HDV poses an important community health problem.

Characterization of proteins associated with hepatitis delta virus.

The number and size of proteins associated with hepatitis delta virus (HDV) from serum and liver (human, chimpanzee and woodchuck) in the acute and chronic stages of HDV infection were analysed by immunoblotting. HDV particles in serum were separated from serum proteins by gel filtration and peak fractions of HDV antigens were subjected to PAGE. Immunoblotting with human anti-HDV-positive sera and 125I-labelled Protein A revealed two bands of 27K and 29K. It was not possible to identify any core-like structure from liver homogenates by CsCl gradient centrifugation. HDV proteins from such gradients were degraded to a size of 14K as determined by immunoblotting. HDV RNA was found in fractions at a density of 1.5 g/ml. However, direct homogenization of liver tissue in gel electrophoresis sample buffer, followed by PAGE and immunoblotting resulted in identification of HDV-associated proteins of 27K and 29K, indicating that HDV proteins in liver tissue are the same size as those in serum, but that they degrade rapidly. There was no difference in size of HDV proteins in liver samples from humans, chimpanzees or woodchucks.

Interleukins in chronic active hepatitis B. Relationship with viral markers.

Interleukin-2, a product of helper T cells, is essentially involved in the regulation of cell-mediated immunity. Two monocyte-derived factors, interleukin-1 and prostaglandin E2, influence interleukin-2 synthesis with opposite actions. To analyse immunoregulatory function in HBsAg-positive chronic active hepatitis, T cell subsets in peripheral blood and the levels of interleukin-2, interleukin-1 and prostaglandin E2 in supernatants from lectin- or lipopolysaccharide-activated peripheral mononuclear cell cultures were determined in 16 healthy controls and 33 patients with chronic active hepatitis B. Interleukin-2 activity was comparable to the controls in patients without delta infection who had seroconverted to anti-HBe (group 1), but it was significantly reduced in both HBeAg-positive subjects (group 2) (P less than 0.05 vs. controls and group 1) and those cases with positive delta markers (group 3) (P less than 0.01 and less than 0.05 vs. controls and group 1, respectively). In group 3, interleukin-2 was similarly diminished in both anti-HTLV-III-positive and -negative cases as well as in HBeAg- and anti-HBe-positive subjects. Notwithstanding the changes in interleukin-2 activity, no significant differences in the number of T4 cells, or in the levels of either interleukin-1 or prostaglandin E2, were found among the various groups of subjects studied. However, in those groups with reduced interleukin-2 activity an increased number of T8 cells was observed. It is suggested that the low levels of interleukin-2 found in the replicative phase of chronic active hepatitis B and in delta superinfection reflect a disturbed immunoregulation that may contribute to persistent viral replication in these two conditions.

Rapid procedure for the detection of hepatitis delta virus RNA in sera of HBsAg-positive and anti-delta-positive patients.

A synthetic oligonucleotide duplex of 78 bp corresponding to part of the recently published RNA sequence of hepatitis Delta virus (HDV) was cloned into the plasmid pSBO1 and used for the detection of HDV RNA in sera of patients with chronic HDV infection by molecular hybridization. RNA containing the 78 bp sequence was synthesized in vitro and used as a positive control. For this purpose, a fragment containing the cloned oligonucleotide was transferred into the plasmid pSPT 18. HDV RNA was present in 5 out of 32 hepatitis B surface antigen (HBsAg)- and anti-HD-positive patients. It was neither found in the sera of 19 HBsAg-positive, anti-HD-negative patients, nor in the sera of 26 patients with chronic liver disease negative for both HBsAg and anti-HD. The method appears to be suitable for the detection of viruses of which either only parts of the genome or the entire sequence is known.

Delta agent and the etiology of hepatocellular carcinoma.

Sera from 87 patients with hepatitis B surface antigen (HBsAg)-positive hepatocellular carcinoma (HCC) and from 29 HBsAg-positive hospital controls were tested for delta (delta) antigen with an immunoenzymatic procedure and for anti-delta antibody, hepatitis Be antigen (HBeAg) and antibody to HBeAg (anti-HBe) by radioimmunoassay. All the sera, from both the HCC cases and the control patients, were negative for delta-antigen. Among the HCC cases 9 were positive for serum anti-delta (10%) whereas among the controls none was positive for this antibody (p = 0.067); the anti-delta-positive cases were found only among HCC patients negative for HBeAg. The lower prevalence of anti-delta among HBsAg-positive HCC patients, as compared to the corresponding prevalence among HBsAg-positive patients with chronic active hepatitis or cirrhosis (reported in the literature) indicates that the pathogenesis of HCC is frequently independent of the pathogenesis of the other HBsAg-positive common chronic liver diseases. By contrast, the higher prevalence of anti-delta among HBsAg-positive HCC cases than among HBsAg-positive controls may reflect the longer average duration of the carrier state in HCC patients (until integration is accomplished and the induction period completed). Serum HBeAg was higher among HBsAg-positive HCC patients with cirrhosis (23%) than among HBsAg-positive HCC patients without cirrhosis (6%) or HBsAg-positive controls (3%); thus, the conflicting results in the literature concerning the association of the HBeAg/anti-HBe system with HCC may be accounted for, in part, by the variable association of HCC with an underlying cirrhosis.

Hepatitis delta virus: protein composition of delta antigen and its hepatitis B virus-derived envelope.

Hepatitis delta virus (HDV)-associated particles were purified from the serum of an experimentally infected chimpanzee by size chromatography and by density centrifugation. Hepatitis delta antigen (HDAg) was detected after mild detergent treatment at a column elution volume corresponding to 36-nm particles and banded at a density of 1.25 g/ml. The serum had an estimated titer of 10(9) to 10(10) HDV-associated particles and had only a 10-fold excess of hepatitis B surface antigen (HBsAg) not associated with HDAg. Therefore, HDV appears to be much more efficiently packed and secreted than is its helper virus, hepatitis B virus (HBV), which is usually accompanied by a 1,000-fold excess of HBsAg. The protein compositions of the HDAg-containing particles were analyzed by immunoblotting with HDAg-, HBsAg-, and hepatitis B core antigen-specific antisera and monoclonal antibodies to HBV surface gene products. The HBsAg envelope of HDAg contained approximately 95% P24/GP27s, 5% GP33/36s, and 1% P39/GP42s proteins. This protein composition was more similar to that of the 22-nm particles of HBsAg than to that of complete HBV. The significant amount of GP33/36s suggests that the HBsAg component of the HDV-associated particle carries the albumin receptor. Two proteins of 27 and 29 kilodaltons which specifically bound antibody to HDAg but not HBV-specific antibodies were detected in the interior of the 36-nm particle. Since these proteins were structural components of HDAg and were most likely coded for by HDV, they were designated P27d and P29d.

Marcadores sorológicos do virus da hepatite B e anticorpo delta em homossexuais masculinos brasileiros / Serological markers for hepatitis B virus and delta antibody in Brazilian male homosexuals

Os marcadores sorológicos do vírus B da hepatite B foram estudados em um grupo de homens homossexuais em Säo Paulo, Brasil. Encontraram-se 23% de positividade para o AgHBs nos 26 homossexuais, sendo que 81% apresentavam positividade para alguns dos marcadores do vírus B. Anticorpo anti-o näo foi detectado em nenhum caso. No grupo controle, encontrou-se 1% de positividade para o AgHBs entre 500 homens estudados