Advertisement

Gastroenterologia Japonica

, Volume 27, Issue 6, pp 734–744 | Cite as

Electron microscopic study of hepatitis B virus-associated antigens on the infected liver cell membrane in relation to analysis of immune target antigens in chronic hepatitis B

  • Takafumi Saito
  • Tomoteru Kamimura
  • Masamichi Ishibashi
  • Haruhide Shinzawa
  • Tsuneo Takahashi
Liver, Pancreas and Biliary Tract
  • 52 Downloads

Summary

To clarify the characteristics of cell surface Pre-S2 Ag, HBcAg and HBeAg immunohistochemically and to explore their relationship with a cellular immune target antigen, 31 liver biopsy specimens from chronic HBV carriers were examined by immunoperoxidase staining. By immune light microscopy, Pre-S2 Ag was detected on the liver cell membrane in 18 (58%) of the 31 cases, HBcAg in 4 cases (13%) and HBeAg in 4 cases (13%). Pre-S2 Ag frequently showed a honeycomb-like membrane expression pattern which was present regardless of liver inflammation, whereas HBcAg and HBeAg exhibited a scattered membrane expression pattern detected in areas of marked inflammation. Of the 18 cases showing a honeycomb-like Pre-S2 Ag expression, 3 concomitantly showed a scattered membrane expression pattern. Immunoelectron microscopy revealed these two distinct membrane expression patterns. In areas showing a honeycomb-like membrane expression pattern, Pre-S2 Ag was demonstrated in the intercellular space and on the basolateral membranes of hepatocytes, but was not detected on the cell membranes in areas of the intercellular space lacking an immunoreaction. Cytoplasmic expression of Pre-S2 Ag was less extensive in these hepatocytes. These findings suggest that the honeycomb-like membrane expression of Pre-S2 Ag results from attachment of extracellular antigen to the liver cell membrane. In contrast, in areas showing a scattered membrane expression pattern, Pre-S2 Ag, HBcAg and HBeAg were each detected as single-layered linear deposits along the cell membrane, but were absent in the intercellular space. Each antigen was also expressed abundantly in the cytoplasm, and the immunoproducts appeared to fuse with the cell membrane. These findings suggest that the scattered membrane expression of these antigens results from intrahepatic transfer of antigen synthesized in the liver cell to the cell membrane, possibly serving as a target for the host immune-mediated response in connection with inflammation.

Key words

hepatitis B hepatitis B virus immunohistochemistry target antigen 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Ferrari C, Penna A, DegliAntoni A, et al. Cellular immune response to hepatitis B virus antigens (An overview). J Hepatol 1988;7:21–33.PubMedCrossRefGoogle Scholar
  2. 2.
    Eggink HF, Houthoff HJ, Huitema S, et al. Cellular and humoral immune reactions in chronic active liver disease. Lymphocyte subsets and viral antigens in liver biopsies of patients with acute and chronic hepatitis B. Clin exp Immunol 1984;56:121–128.PubMedGoogle Scholar
  3. 3.
    Thomas HC, Path MRC, Lok ASF. The immunopathology of autoimmune and hepatitis B virus-induced chronic hepatitis. Sem Liver Dis 1984;4:36–46.CrossRefGoogle Scholar
  4. 4.
    Mondelli M, Eddleston ALWF: Mechanisms of liver cell injury in acute and chronic hepatitis B. Sem Liver Dis 1984;4:47–58.Google Scholar
  5. 5.
    Eddleston ALWF, Mondelli M. Immunopathological mechanisms of liver cell injury in chronic hepatitis B virus infection. J Hepatol 1986;3(Suppl. 2):sl7-s23.Google Scholar
  6. 6.
    Bianchi L, Groote JD, Desmet VJ, et al. Acute and chronic hepatitis revisited. Review by an international group. Lancet 1977 ;914–919.Google Scholar
  7. 7.
    Saito T, Kamimura T, Asakura H, et al. Ultrastructural localization of Pre-S2 polypeptides in the liver tissues of patients with chronic hepatitis B virus infection. Liver 1989;9:329–337.PubMedGoogle Scholar
  8. 8.
    Alberti A, Realdi G, Bortolotti F, et al. T-lymphocyte cytotoxicity to HBsAg-coated target cells in hepatitis B virus infection. Gut 1977;18:1004–1009.PubMedCrossRefGoogle Scholar
  9. 9.
    Mondelli M, Vergani GM, Alberti A, et al. Specificity of T lymphocyte cytotoxicity to autologous hepatocytes in chronic hepatitis B virus infection: evidence that T cells are directed against HBV core antigen expressed on hepatocytes. J Immunol 1982;129: 2773–2778.PubMedGoogle Scholar
  10. 10.
    Naumov NV, Mondelli M, Alexander GJM, et al. Relationship between expression of hepatitis B virus antigens in isolated hepatocytes and autologous lymphocyte cytotoxicity in patients with chronic hepatitis B virus infection. Hepatology 1984;4:63–68.PubMedGoogle Scholar
  11. 11.
    Vento S, Hegarty JE, Alberti A, et al. T lymphocyte sensitization to HBcAg and T cell-mediated unresponsiveness to HBsAg in hepatitis B virus-related chronic liver disease. Hepatology 1985; 5:192–197.PubMedCrossRefGoogle Scholar
  12. 12.
    Mondelli M, Bortolotti F, Pontisso P, et al. Definition of hepatitis B virus (HBV)-specific target antigens recognized by cytotoxic T cells in acute HBV infection. Clin Exp Immunol 1987;68:242–250.PubMedGoogle Scholar
  13. 13.
    Ray MB, Desmet VJ, Bradburne AF, et al. Differential distribution of hepatitis B surface antigen and hepatitis B core antigen in the liver of hepatitis B patients. Gastroenterology 1976;71:462–467.PubMedGoogle Scholar
  14. 14.
    Chu CM, Liaw YF. Intrahepatic distribution of hepatitis B surface and core antigens in chronic hepatitis B virus infection. Gastroenterology 1987;92:220–225.PubMedGoogle Scholar
  15. 15.
    Hsu HC, Su IJ, Lai MY, et al. Biologic and prognostic significance of hepatocyte hepatitis B core antigen expressions in the natural course of chronic hepatitis B virus infection. J Hepatol 1987;5:45–50.PubMedCrossRefGoogle Scholar
  16. 16.
    Suzuki K, Uchida T, Shikata T. Histopathological analysis of chronic hepatitis B virus (HBV) infection in relation to HBV replication. Liver 1987;7:260–270.PubMedGoogle Scholar
  17. 17.
    Trevisan A, Realdi G, Alberti A, et al. Core antigen-specific immunoglobulin G bound to the liver cell membrane in chronic hepatitis B. Gastroenterology 1982;82:218–222.PubMedGoogle Scholar
  18. 18.
    Ramalho F, Brunetto M, Rocca G, et al. Serum markers of hepatitis B virus replication, liver histology and intrahepatic expression of hepatitis B core antigen. J Hepatol 1988;7:14–20.PubMedCrossRefGoogle Scholar
  19. 19.
    Kojima T, Bloemen J, Desmet VJ. Immune electron microscopic demonstration of hepatitis B core antigen (HBcAg) in liver cell plasma membranes. Liver 1987;7:191–200.PubMedGoogle Scholar
  20. 20.
    Sasaki H, Kojima T, Matsui S, et al. Interaction of lymphocytes with hepatocytes containing hepatitis B antigen: ultrastructural demonstration of target antigen and T-cell subsets by the peroxidase antibody technique. Virchows Arch A 1987;411:489–498.CrossRefGoogle Scholar
  21. 21.
    Aoyama K, Kojima T, Inoue K, et al. Immunohistochemical investigation of hepatitis B virus associated antigens, HLA antigens and lymphocyte subsets in type B chronic hepatitis. Gastroenterol Jpn 1990;25:41–53.PubMedGoogle Scholar
  22. 22.
    Barnaba V, Franco A, Alberti A, et al. Recognition of hepatitis B virus envelope proteins by liver-infiltrating T lymphocytes in chronic HBV infection. J Immunol 1989;143:2650–2655.PubMedGoogle Scholar
  23. 23.
    Hu KQ, Hao LJ, Zhang YY, et al. Intrahepatic Expression of Pre-S proteins of hepatitis B virus and its possible relation to liver cell necrosis. Am J Gastroenterol 1989;84:1538–1542.PubMedGoogle Scholar
  24. 24.
    Pignatelli M, Waters J, Lever A, et al. Cytotoxic T-cell responses to the nucleocapsid proteins of HBV in chronic hepatitis. Evidence that antibody modulation may cause protracted infection. J Hepatol 1987;4:15–21.PubMedCrossRefGoogle Scholar
  25. 25.
    Milich DR, McLachlan A, Moriarty A, et al. Immune response to hepatitis B virus core antigen (HBcAg): Localization of T cell recognition sites within HBcAg/HBeAg. J Immunol 1987;139: 1223–1231.PubMedGoogle Scholar
  26. 26.
    Kakumu S, Arao M, Mizokami M, et al. Pre-S proteins in chronic hepatitis B virus infection: markers of active viral infection. Am J Gastroenterol 1989;84:1250–1254.PubMedGoogle Scholar
  27. 27.
    Yamada G, Takaguchi K, Matsueda K, et al. Immunoelectron microscopic observation of intrahepatic HBeAg in patients with chronic hepatitis B. Hepatology 1990;12:133–140.PubMedCrossRefGoogle Scholar
  28. 28.
    Budkowska A, Dubreil P, Capel F, et al. Hepatitis B virus pre-S2 gene-encoded antigenic specificity and anti-pre-S2 antibody: Relationship between anti-pre-S2 response and recovery. Hepatology 1986;6:360–368.PubMedCrossRefGoogle Scholar
  29. 29.
    Budkowska A, Riottot MM, Dubreuil P, et al. Monoclonal antibody recognizing pre-S2 epitope of hepatitis B virus: Characterization of pre-S2 epitope and anti-pre-S2 antibody. J Med Virol 1986;20:111–125.PubMedCrossRefGoogle Scholar
  30. 30.
    Okamoto H, Imai M, Usuda S, et al. Hemagglutination assay of polypeptide coded by the pre-S region of hepatitis B virus DNA with monoclonal antibody; correlation of pre-S polypeptide with the receptor for polymerized human serum albumin in serums containing hepatitis B antigens. J Immunol 1985;134:1212–1216.PubMedGoogle Scholar

Copyright information

© The Japanese Society of Gastroenterology 1992

Authors and Affiliations

  • Takafumi Saito
    • 1
  • Tomoteru Kamimura
    • 2
  • Masamichi Ishibashi
    • 1
  • Haruhide Shinzawa
    • 1
  • Tsuneo Takahashi
    • 1
  1. 1.Second Department of Internal MedicineYamagata University School of MedicineYamagataJapan
  2. 2.Third Department of Internal MedicineNiigata University School of MedicineNiigataJapan

Personalised recommendations