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Clinical and Experimental Immunology logoLink to Clinical and Experimental Immunology
. 1989 Jan;75(1):64–69.

Relationship between the intrahepatic expression of 'e' and 'c' epitopes of the nucleocapsid protein of hepatitis B virus and viraemia.

M Ballaré 1, C Lavarini 1, M R Brunetto 1, E Petruzzelli 1, M Dovis 1, G Molino 1, F Bonino 1
PMCID: PMC1541863  PMID: 2467769

Abstract

The relationship between hepatitis B viraemia and intrahepatic HBV nucleocapsid proteins (HBcAg and HBeAg) was studied in 18 patients with chronic hepatitis B. Monoclonal antibodies (MoABs) were obtained in BALB/c mice primed with recombinant HBV nucleocapsid proteins. Four MoABs reacting with recombinant proteins gave positive results in competitive assays. Two reacted as anti-HBc and two as anti-HBe. One of them showed a strong affinity for the cytoplasmic, membrane-bound antigen (P23e) of infected hepatocytes while the latter showed a higher specificity for serum HBeAg than for the intrahepatic antigen. Anti-HBc MoABs had a staining capacity for liver cell nuclei comparable with that of polyclonal antibodies. Overall the anti-HBc MoABs stained the liver cell nuclei in 86% of cases, while anti-HBe MoABs stained in 58% of cases. The hepatocyte cytoplasm was stained by anti-HBc MoABs and anti-HBe MoABs in 64% and 72% of cases respectively. Not one of 12 control liver biopsies was stained. Viraemia (HBV-DNA) was measured by dot blot hybridization and was correlated with the number of hepatocytes containing the nucleocapsid antigen. The highest levels of HBV-DNA (greater than 10(8) genomes/ml) were detected in patients with prevalent nuclear staining while the lowest ones were observed in those with prevalent cytoplasmic expression of this antigen. The application of anti-HBV-nucleocapsid MoABs in diagnostics requires careful scrutiny since some are specific for the circulating antigen while others show a higher affinity for the intrahepatic antigen.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bonino F., Hoyer B., Nelson J., Engle R., Verme G., Gerin J. Hepatitis B virus DNA in the sera of HBsAg carriers: a marker of active hepatitis B virus replication in the liver. Hepatology. 1981 Sep-Oct;1(5):386–391. doi: 10.1002/hep.1840010503. [DOI] [PubMed] [Google Scholar]
  2. Bonino F., Rosina F., Rizzetto M., Rizzi R., Chiaberge E., Tardanico R., Callea F., Verme G. Chronic hepatitis in HBsAg carriers with serum HBV-DNA and anti-HBe. Gastroenterology. 1986 May;90(5 Pt 1):1268–1273. doi: 10.1016/0016-5085(86)90395-1. [DOI] [PubMed] [Google Scholar]
  3. Boniolo A., Dovis M., Matteja R. Use of an enzyme-linked immunosorbent assay for screening hybridoma antibodies against hepatitis B surface antigen. J Immunol Methods. 1982;49(1):1–15. doi: 10.1016/0022-1759(82)90361-1. [DOI] [PubMed] [Google Scholar]
  4. Goding J. W. Antibody production by hybridomas. J Immunol Methods. 1980;39(4):285–308. doi: 10.1016/0022-1759(80)90230-6. [DOI] [PubMed] [Google Scholar]
  5. Gowans E. J., Burrell C. J., Jilbert A. R., Marmion B. P. Cytoplasmic (but not nuclear) hepatitis B virus (HBV) core antigen reflects HBV DNA synthesis at the level of the infected hepatocyte. Intervirology. 1985;24(4):220–225. doi: 10.1159/000149646. [DOI] [PubMed] [Google Scholar]
  6. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  7. MacKay P., Lees J., Murray K. The conversion of hepatitis B core antigen synthesized in E coli into e antigen. J Med Virol. 1981;8(4):237–243. doi: 10.1002/jmv.1890080404. [DOI] [PubMed] [Google Scholar]
  8. Miller R. H. Proteolytic self-cleavage of hepatitis B virus core protein may generate serum e antigen. Science. 1987 May 8;236(4802):722–725. doi: 10.1126/science.3554507. [DOI] [PubMed] [Google Scholar]
  9. Mondelli M., Tedder R. S., Ferns B., Pontisso P., Realdi G., Alberti A. Differential distribution of hepatitis B core and E antigens in hepatocytes: analysis by monoclonal antibodies. Hepatology. 1986 Mar-Apr;6(2):199–204. doi: 10.1002/hep.1840060208. [DOI] [PubMed] [Google Scholar]
  10. Nakane P. K., Kawaoi A. Peroxidase-labeled antibody. A new method of conjugation. J Histochem Cytochem. 1974 Dec;22(12):1084–1091. doi: 10.1177/22.12.1084. [DOI] [PubMed] [Google Scholar]
  11. Negro F., Chiaberge E., Oliviero S., Hammer M., Berninger M., Canese M. G., Bonino F. Hepatitis B virus DNA (HBV-DNA) in anti-HBe positive sera. Liver. 1984 Jun;4(3):177–183. doi: 10.1111/j.1600-0676.1984.tb00925.x. [DOI] [PubMed] [Google Scholar]
  12. Seeger C., Ganem D., Varmus H. E. Biochemical and genetic evidence for the hepatitis B virus replication strategy. Science. 1986 Apr 25;232(4749):477–484. doi: 10.1126/science.3961490. [DOI] [PubMed] [Google Scholar]
  13. Takahashi K., Machida A., Funatsu G., Nomura M., Usuda S., Aoyagi S., Tachibana K., Miyamoto H., Imai M., Nakamura T. Immunochemical structure of hepatitis B e antigen in the serum. J Immunol. 1983 Jun;130(6):2903–2907. [PubMed] [Google Scholar]
  14. Tiollais P., Pourcel C., Dejean A. The hepatitis B virus. Nature. 1985 Oct 10;317(6037):489–495. doi: 10.1038/317489a0. [DOI] [PubMed] [Google Scholar]
  15. Uy A., Bruss V., Gerlich W. H., Köchel H. G., Thomssen R. Precore sequence of hepatitis B virus inducing e antigen and membrane association of the viral core protein. Virology. 1986 Nov;155(1):89–96. doi: 10.1016/0042-6822(86)90170-4. [DOI] [PubMed] [Google Scholar]

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