Intra- and extracellular distribution and immunochemical characterization of hepatitis B virus nucleocapsid proteins produced by a human hepatoma cell line transfected with cloned viral DNA

Ohori, Hitoshi; Matsuda, Keizaburoh

doi:10.1016/0042-6822(89)90401-7

Cited by 7 publications

(1 citation statement)

References 26 publications

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“…The precore gene product excreted in the medium was heterogenous and consisted of four discrete polypeptides between 15 k and 19 k in molecular weight. Similar results have been obtained when the precore gene was expressed by other vector systems [3,16,32,37,41], where the heterogeneity was attributed to possible multiple cleavage sites in the arginine-rich carboxyterminus. Recombinant viruses (WyAd7HSPC-8) which contained the S gene, as well as the precore gene, expressed similar amounts of HBeAg compared to WyAd7HPC-3 but the product found by Western blotting seemed less heterogenous.…”

Section: Discussionsupporting

confidence: 85%

Co-expression of hepatitis B virus antigens by a non-defective adenovirus vaccine vector

Ye¹,

Mason²,

Chengalvala³

et al. 1991

Archives of Virology

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Adenovirus type 7 vaccine strain was engineered to express foreign antigens from both the E3 early promoter in the E3 region and the major late promoter inserted between the E4 region and the right inverted terminal repeat. This multiple expression vector was used to express hepatitis B core antigen (HBcAg), hepatitis B e antigen (HBeAg), and hepatitis B surface antigen (HBsAg). The gene inserted in the E3 region was derived from the core gene of the hepatitis B virus genome. When the precore region was present, an immunoreactive group of proteins with molecular weights ranging from 15,000 to 19,000 was secreted into the media. Velocity sedimentation centrifugation of media and lysates from cells infected with recombinants containing the core gene with the precore region resulted in peaks of HBeAg at the top of the gradient where authentic HBeAg should be found. In addition to the core gene in the E3 region, the surface antigen gene of hepatitis B virus was inserted behind the major late promoter in the E4 region resulting in an adeno-hepatitis recombinant virus capable of expressing both the core gene and the HBsAg cells. Cells infected with the adeno-hepatitis recombinants could also be stained with peroxidase-conjugates after reacting to antibody against HBcAg. Inoculation of dogs with the recombinant viruses which contained the core gene, with and without the precore sequence, resulted in a significant antibody response to HBcAg/HBeAg. The dogs also produced a significant antibody response to HBsAg as well as neutralizing antibody to adenovirus.

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Section: Discussionsupporting

confidence: 85%

Co-expression of hepatitis B virus antigens by a non-defective adenovirus vaccine vector

Ye¹,

Mason²,

Chengalvala³

et al. 1991

Archives of Virology

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Molecular species of hepadnavirus core and envelope polypeptides in hepatocyte plasma membrane of woodchucks with acute and chronic viral hepatitis

Michalak

Lin

1994

Hepatology

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Hepadnaviral antigens exposed on hepatocytes serve as targets and as possible modulators of immunopathogenic reactions causing liver damage. To identify molecular species of viral proteins at the liver cell surface and the relationship between their expression and the persistence and severity of virus-induced liver injury, we examined woodchuck hepatitis virus core and envelope polypeptides in host hepatocyte plasma membranes from acute and chronic hepatitis. Western blot analysis revealed that two virus core polypeptides with a molecular mass of 22 kD and 43 kD occur in the membranes of infected animals. The molecular profiles and the membrane levels of the core antigen were not related to the duration or histological severity of liver damage. In contrast, quantities of the virus surface antigen were significantly greater in hepatocyte membranes of animals with chronic hepatitis. The envelope preS1, preS2 and S polypeptides, with or without molecular mass equivalents in the subviral surface antigen particles and virions, were detected in all infected membranes, although the preS2 polypeptides were always dominant. Our findings indicate that hepadnavirus core and envelope polypeptides are integral constituents of hepatocyte membranes in the course of hepatitis. They demonstrate that the accumulation of viral envelope proteins, predominantly the S-domain sequences, in hepatocyte membranes is a prominent characteristic of chronic hepatitis. This event might contribute to promotion of development and maintenance of hepadnavirus-related chronic liver disease.

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Pathomorphological characteristics and pathogenesis of viral hepatitis

Schaff

Lotz²,

Schulte-Herman³

1996

Pathol. Oncol. Res.

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Viral hepatitis (VH) is an inflammatory reaction of the liver to hepatotropic viruses. Acute VH can be classified according to the virus and type of necrosis. Chronic hepatitis (CH) might be active, persistent or lobular based on previous classification. More recently the grade (necroinflammatory activity) and stage (fibrosis and architectural distorsion) of CH have been distinguished and scored. Apoptosis and necrosis probably coexist in VH and contribute to hepatocyte death. Several "death factors", such as transforming growth factor b, Apo1/Fas and tumor necrosis factor play a role in the execution of cell death. Injury of hepatocytes during viral infection can occur as a direct effect of the virus or as a result of the host immune response. Expression of different viral antigens can be detected during VH and might be visualized. Phenotyping of the portal inflammatory cell infiltrate in CH has shown a T-cell zone comprised of CD4+ helper T cells and CD8+ supressor/cytotoxic T cells at the periphery of the lobules. The pathogenetic mechanisms responsible for the final outcome of viral infection depend on viral factors (such as genotype, mutation etc.), virus-host interaction, expression of viral protein, several cytokines etc. which finally lead to the well known histological alterations of viral hepatitis.

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Intra- and extracellular distribution and immunochemical characterization of hepatitis B virus nucleocapsid proteins produced by a human hepatoma cell line transfected with cloned viral DNA

Cited by 7 publications

References 26 publications

Co-expression of hepatitis B virus antigens by a non-defective adenovirus vaccine vector

Co-expression of hepatitis B virus antigens by a non-defective adenovirus vaccine vector

Molecular species of hepadnavirus core and envelope polypeptides in hepatocyte plasma membrane of woodchucks with acute and chronic viral hepatitis

Pathomorphological characteristics and pathogenesis of viral hepatitis

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