Virchows Archiv A

, Volume 411, Issue 5, pp 489–498 | Cite as

Interaction of lymphocytes with hepatocytes containing hepatitis B antigen: Ultrastructural demonstration of target antigen and T-cell subsets by the peroxidase antibody technique

  • Hiroshi Sasaki
  • Takashi Kojima
  • Shinjiro Matsui
  • Keiichi Aoyama
  • Kyoichi Inoue
Article
Accepted:

Summary

Hepatitis B virus associated antigens and subsets of lymphocytes in liver tissue were studied using immune electron microscopy to clarify the immune mechanism of hepatocyte lysis in type B chronic hepatitis. Using conventional electron microscopy, infiltrating lymphocytes were observed in direct contact with hepatocytes in areas of piecemeal necrosis and focal necrosis; they showed various types of surface adherence with a contact gap of approximately 20 nm in width. The majority of the hepatocytes that were in contact with lymphocytes could be shown to contain HBsAg and/or HBcAg by immune electron microscopy: HBsAg was localized in the endoplasmic reticulum membranes, in tubular structures, and on the outer coat of Dane particles; HBcAg was observed in the nuclei and in the cytoplasmic matrix of hepatocytes. In some cases HBsAg was observed on the plasma membrane of hepatocyte in contact with lymphocytes. Immune electron microscopy using monoclonal antibodies to subsets of human T-lymphocytes revealed that the lymphocytes in areas of piecemeal necrosis and focal necrosis were predominantly CD 5 or CD 8 positive. In contrast, CD 4 positive cells were infrequently observed in necro-inflammatory regions and Leu 7 positive cells were randomly scattered in the sinusoids away from areas of hepatocyte necrosis. These data suggest that HBsAg is at least one of the target antigens expressed on the hepatocyte membrane possibly enabling cytolytic interaction by cytotoxic T cells in chronic type B hepatitis.

Key words

Immune electron microscopy T-cell subsets HBV associated antigens Lymphocyte interaction 
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References

  1. Alberti A, Realde G, Bortolotti F, Rigoli AM (1977) T-lymphocyte cytotoxicity to HBsAg-coated target cells in hepatitis B virus infection. Gut 18:1004–1009Google Scholar
  2. Bernuau D, Rogier E, Feldmann G (1982) A quantitative ultrastructural analysis of the leucocytes in contact with hepatocytes in chronic active hepatitis, with a cytochemical detection of mononuclear phagocytes. Am J Pathol 109:310–320Google Scholar
  3. Biberfeld P, Johannsson A (1975) Contact areas of cytotoxic lymphocytes and target cells: An electron microscopic study. Exp Cell Res 94:79–87Google Scholar
  4. Burns WH, Allison AC (1977) Surface antigens of virus-infected cells. In: Poste G, Nicolson GL (eds) Virus infection and the cell surface North-Holland, Amsterdam, pp 213–247Google Scholar
  5. Eddleston ALWF, Mondelli M, Mieli-Vergani G, Williams R (1982) Lymphocyte cytotoxicity to autorogous hepatocytes in chronic hepatitis B virus infection. Hepatorogy 2:122S-127SGoogle Scholar
  6. Elsheikh N, Osman CG, Cullens H, Eddleston ALWF, Williams R (1978) T lymphocyte-mediated cytotoxicity in HBsAg-positive liver disease. Clin Exp Immunol 31:158–165Google Scholar
  7. Galbraith RM, Fudenberg HH (1977) A review. Autoimmunity in chronic active hepatitis and diabetes mellitus. Clin Immunol Immunopathol 18:116–149Google Scholar
  8. Govindarajan S, Uchida T, Peters RL (1983) Identification of T lymphocytes and subsets in liver biopsy cores of acute viral hepatitis. Liver 3:13–18Google Scholar
  9. Hadziyannis S, Gerber MA, Vissoulis C, Popper H (1973) Cytoplasmic hepatitis B antigen in “ground glass” hepatocytes of carriers. Arch Pathol 96:327–330Google Scholar
  10. Henny CS (1973) Studies on the mechanism of lymphocyte-mediated cytotoxicity. II. The use of various target cell markers to study cytolytic events. J Immunol 110:73–84Google Scholar
  11. Hopf U, Meyer zum Büschenfelde KH, Arnold W (1976) Detection of a liver-membrane autoantibody in HBsAg-negative chronic active hepatitis. N Engl J Med 294:578–582Google Scholar
  12. Hütteroth TH, Poralla T, Meyer zum Büschenfelde KH (1981) Spontaneous cell-mediated (SCMC) and antibody-dependent cellular cytotoxicity (ADCC) in patients with acute and chronic antive hepatitis. Klin Wochenschr 59:699–706Google Scholar
  13. Kawanishi H (1977) Morphologic association of lymphocytes with hepatocytes in chronic liver disease. Arch Pathol Lab Med 101:286–290Google Scholar
  14. Kojima T (1982) Immune electron microscopic study of hepatitis B virus associated antigens in hepatocytes. Gastroent Jpn 17:558–575Google Scholar
  15. Kojima T, Desmet VJ (1984) Hepatitis B core antigen (HBcAg) in liver cell plasma membrane: Immunoelectron microscopic study. Hepatology 4:780Google Scholar
  16. Koren HS, Ax W, Elisabeth FM (1973) Morphological observations on the contact-induced lysis of target cell. Eur J Immunol 3:32–37Google Scholar
  17. Liepins A, Faanes RB, Lifter J, Choi YS, De Harven E (1977) Ultrastructural changes during T-lymphocyte-mediated cytolysis. Cell Immunology 28:109–124Google Scholar
  18. Matter A (1979) Microcinematographic and electron microscopic analysis of target cell lysis induced by cytotoxic T lymphocytes. Immunology 36:179–190Google Scholar
  19. McLean IW, Nakane PK (1974) Periodate-lysine-paraformal-dehyde fixative: a new fixative for immunoelectron microscopy. J Histochem Cytochem 22:1077–1083Google Scholar
  20. Meyer zum Büschenfelde KH, Hütteroth TH, Arnold W, Hopf U (1979) Immunologic liver injury: The role of hepatitis B viral antigens and liver membrane antigens as targets. In: Popper H, Schaffner F, eds. Progress in liver diseases, Vol VI. Grune and Stratton, New York, pp 407–424Google Scholar
  21. Mondelli M, Mieli-Vergani G, Alberti A, Vergani D, Portmann B, Eddleston ALWF, Williams R (1982) 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 129:2273–2778Google Scholar
  22. Montano L, Arguibel F, Boffill M, Goodall AH, Janossy G, Thomas HC (1983) An analysis of the inflammatory infiltrate in autoimmune and hepatitis B virus-induced chronic liver disease. Hepatology 3:292–296Google Scholar
  23. Nakane PK, Kawaoi A (1974) Peroxidase-labeled antibody. A new method of conjugation. J Histochem Cytochem 22:1084–1091Google Scholar
  24. Pape GR, Rieber EP, Eisenburg J (1983) Involvement of the cytotoxic/suppressor T-cell subset in liver tissue injury of patients with acute and chronic liver diseases. Gastroenterology 85:657–662Google Scholar
  25. Ray MB, Desmet VJ, Fevery J, De Groote J, Bradburne AF, Desmyter J (1976) Distribution patterns of hepatitis B surface antigen (HBsAg) in the liver of hepatitis patients. J Clin Pathol 29:94–100Google Scholar
  26. Review by an International Group (1971) Morphologic criteria in viral hepatitis. Lancet I:333–337Google Scholar
  27. Review by an International Group (1977) Acute and chronic hepatitis revisited. Lancet II:914–919Google Scholar
  28. Russell JH, Masakowski VR, Dobos CB (1980) Mechanisms of immune lysis. I. Physiological distinction between target cell death mediated by cytotoxic T lymphocytes and antibody plus complement. J Immunol 124:1100–1105Google Scholar
  29. Russell JH, Dobos CB (1980) Mechanisms of immune lysis. II. CTL-induced nuclear disintegration of the target begins within minutes of cell contact. J Immunol 125:1256–1261Google Scholar
  30. Sanderson CJ, Glauert AM (1977) The mechanism of T-cell mediated cytotoxicity. V. Morphological studies by electron microscopy. Proc R Soc Lond B 198:315–323Google Scholar
  31. Sasaki H, Ichida F, Kamimura T, Kojima T (1981) Ultrastructural study of interaction of lymphocytes and hepatocytes bearing hepatitis B antigen. In: Berk PD, Chalmers TC (eds) Frontiers in liver disease. Thieme-Stratton, New York, pp 171–176Google Scholar
  32. Sasaki H, Kojima T, Kamimura T, Ichida F (1982) Immunoelectron microscopic study of interaction of lymphocytes and hepatocytes bearing hepatitis B virus (HBV) related antigens Abstract. Hepatology 2:167Google Scholar
  33. Sasaki H, Kojima T, Matsui S, Aoyama K, Higuchi K, Inoue K (1984) Interaction of lymphocytes with hepatocytes bearing hepatitis B antigen: Ultrastructural investigation of target antigen and T cell subsets by means of peroxidase antibody technique Abstract. Hepatology 4:781Google Scholar
  34. Si L, Whiteside TL, Van Thiel, D.H., Rabin BS (1984) Lymphocyte subpopulations at the site of “piecemeal” necrosis in end stage chronic liver diseases and rejecting liver allografts in cyclosporine-treated patients. Lab Invest 50:341–347Google Scholar
  35. Trevisan A, Realdi G, Alberti A, Ongaro G, Pornaro E, Meliconi R (1982) Core antigen-specific immunoglobulin-G bound to the liver cell membrane in chronic hepatitis B. Gastroenterology 82:218–222Google Scholar
  36. Warnatz H, Rösch W, Gerlich W, Gutmann W (1979) Antibody-dependent cell-mediated cytotoxicity (ADCC) and cell-mediated cytotoxicity (CMC) to HBsAg-coated target cells in patients with hepatitis B and chronic hepatitis (CAH). Clin Exp Immunol 35:133–140Google Scholar
  37. Yamada G, Nishihara T, Hyodo I, Manabe K, Fujiki S, Okushin H, Mizuno M, Konoyama S, Kobayashi T, Nagashima H (1984) Cellular immune response in liver of patients with chronic hepatitis B: electron microscopic observation of lymphocyte subsets by the immunoperoxidase method using monoclonal antibodies. Gastroent Jpn 19:517–528Google Scholar

Copyright information

© Springer-Verlag 1987

Authors and Affiliations

  • Hiroshi Sasaki
    • 1
  • Takashi Kojima
    • 1
  • Shinjiro Matsui
    • 1
  • Keiichi Aoyama
    • 1
  • Kyoichi Inoue
    • 1
  1. 1.Third Division, Department of Internal MedicineToyama Medical and Pharmaceutical UniversityToyamaJapan

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