Summary
The diagnostic significance of orcein, aldehydthionine, and chromotrope anilinblue stains for the demonstration of HBsAg containing hepatocytes was investigated in 602 unselected liver biopsies. Five types of specifically stained ground-glass hepatocytes (GGH) were distinguished: Type I showed a positive staining reaction of the cytoplasmic periphery (marginal GGH), type II a diffuse staining of the total cytoplasm (diffuse GGH). Type III contained round or oval globular positive cytoplasmic masses (globular GGH). Type IV showed only very small round, drop-like or sickle-shaped positive structures (spotty GGH). The GGH with fatty changes were designated as type V.
In all carriers and patients with minimal hepatitis GGH, mostly type I and II, appeared in extensive clusters within the lobules. In chronic persistent hepatitis, there were moderately numerous, partly grouped, partly disseminated ground-glass hepatocytes of type II and III. In chronic active hepatitis there were only a few GGH of type IV. In acute viral hepatitis, there were no typical GGH, however, positively stained phagocytes were seen. The intracellular antigen localization and the intralobular distribution of GGH are considered to be the result of an immune reaction.
Single so-called ‘metabolic’ GGH sometimes showed similar pictures. However, they could usually be distinguished from virus containing GGH because of their granular cytoplasmic structure and a lower staining intensity in the applied stains. Among the three stains the orcein stain yielded the best results. In some cases with HBsAg-positive chronic active hepatitis virus infection could not be proved by means of staining.
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Arnold, W., Meyer zum Büschenfelde, K.H., Hess, G., Knolle, J.: The diagnostic significance of intrahepatocellular hepatitis-B-surface-antigen (HBsAG), hepatitis-B-core-antigen (HBcAG) and IgG for the classification of inflammatory liver diseases. (Studies on HBsAG-positive and negative patients). Klin. Wochenschr. 53, 1069–1074 (1975)
Bartók, I., Remenár, É., Tóth, J.: Demonstration of hepatitis B surface antigen by orcein staining in paraffin sections of cirrhotic liver. Virchows Arch. A Path. Anat. Histol. 369, 239–248 (1976)
Bianchi, L., Gudat, F.: Histologische Charakteristika und Nachweis von Hepatitis-B-Komponenten im Lebergewebe bei akuter und chronischer Virushepatitis. Immunität und Infekt 3, 1959–1971 (1975)
Bianchi, L., Gudat, F., Schmid, M.: Semiquantitative correlations between appearance of hepatitis B antigen components in liver and blood in various forms of hepatitis B. In: The liver. Quantitative aspects of structure and function, Preising, R., Bircher, J., Paumgartner, G. (Hrsg.), 2nd Internat. Gstaad Symposium, Gstaad 1975, S. 99. Aulendf.: Editio Cantor 1976
Bianchi, L., Gudat, F.: Hepatitis B: Immunpathologie der verschiedenen Verlaufsformen. Schweiz. Med. Wochenschr. 107, 929–935 (1977)
Bogomoletz, M.V.: Orcein staining of hepatitis B antigen (HBsAG) in conventional paraffin sections of liver biopsies. Acta Hepato-Gastroenterol. 23, 412–414 (1976)
Borchard, F., Gussmann, V.: Vergleichende Untersuchungen mit drei verschiedenen Färbungen zum Nachweis des Australia-Antigens in 602 nicht selektierten Leberpunktatzylindern. Verh. Dtsch. Ges. Path. 61, 476 (1977)
Cohen, C., Berson, S.D., Geddes, E.W.: Hepatitis B antigen in black patients with hepatocellular carcinoma — Correlation between orcein stained liver sections and serology. Cancer 41, 245–249 (1978)
Deodhar, K.P., Tapp, E., Scheuer, P.J.: Orcein staining of hepatitis B antigen in paraffin sections of liver biopsies. J. Clin. Pathol. 28, 66–70 (1975)
Desmet, V.J.: Histopathology of acute and chronic hepatitis. Acta Gastroenterol. Belg. 39, 299–306 (1976)
Endo, Y., Gudat, F., Bianchi, L., Mihatsch, M., Grasser, M., Stalder, G.A., Schmid, M.: Anti-HBc im Rahmen der Hepatitis-B-Virusinfektion: Korrelationen zu Entzündungsform und Virusexpression. Schweiz. Med. Wochenschr. 108, 363–374 (1978)
Gudat, F., Bianchi, L., Sonnabend, W., Thiel, G., Aenishaenslin, W., Stalder, G.A.: Pattern of core and surface expression in liver tissue reflects state of specific immune response in hepatitis B. Lab. Invest. 32, 1 (1975)
Gudat, F., Bianchi, L., Stalder, G.A., Schmid, M.: Klassifizierung und Infektiosität der chronischen Hepatitis B, definiert durch Dane-Partikel im Blut und Viruskomponenten in der Leber. Schweiz. Med. Wochenschr. 106, 812 (1976)
Gudat, F., Bianchi, L.: HBsAG: A target antigen on the liver cell? In: Membrane alterations as basis of liver injury, Popper, H., Bianchi, L., Reuter, W. (Hrsg.) Lancaster: MTP Press 1977a
Gudat, F., Bianchi, L., Finch, M., Krey, G., Endo, Y.: Nuclear fluorescence of liver cells for IgG in viral hepatitis B: Significance and relation to hepatitis B-core and anti-hepatitis B-core formation. Klin. Wochenschr. 55, 329–336 (1977)
Hadziyannis, S., Gerber, M.A., Vissoulis, C., Popper, H.: Cytoplasmic hepatitis B antigen in ‘ground-glass’ hepatocytes of carriers. Arch. Path. 96, 327–330 (1973)
Hinkel, G.K., Probstain, A.-R., Roschlau, G., Stötzner, H., Kemmer, C.: Bedeutung von Milchglashepatozyten im Leberpunktat von Kindern mit chronischer Hepatitis. Dt. Gesundh.-Wesen 32, 1365–1370 (1977)
Hopf, U., Meyer zum Büschenfelde, K.-H., Arnold, W.: Detection of liver-membrane autoantibody in HBsAG-negative chronic active hepatitis. N. Engl. J. Med. 294, 578–582 (1976)
Klinge, O., Bannasch, P.: Zur Vermehrung des glatten endoplasmatischen Retikulums in Hepatozyten menschlicher Leberpunktate. Verh. Dtsch. Ges. Path. 52, 568–573 (1968)
Klinge, O., Kaboth, U., Winckler, K.: Feingewebliche Befunde an der Leber klinisch gesunder Australia-Antigen (HBsAG)-Träger. Virchows Arch. A Path. Anat. Histol. 361, 359–368 (1973)
Klinge, O.: Minimalhepatitis. Verh. Dtsch. Ges. Path. 60, 482 (1976)
Kostich, N.D., Ingham, C.D.: Detection of hepatitis B surface antigen by means of orcein staining of the liver. Am. J. Clin. Path. 67, 20–30 (1977)
Krutsay, M.: Die Markierung der Hepatitis-B-Antigens mit Resorcin-Fuchsin. Orvosi Hetilap 118, 443 (1977)
Ludwig, J., Dickson, E.R., McDonald, G.S.A.: Staging of chronic nonsuppurative destructive cholangitis (Syndrome of primary biliary cirrhosis). Virchows Arch. A Path. Anat. Histol. 379, 103–112 (1978)
Meyer zum Büschenfelde, K.-H., Alberti, A., Arnold, W., Freudenberg, J.: Organ-specificity and diagnostic value of cell-mediated immunity against a liver-specific membrane-protein: Studies in hepatic and non-hepatic diseases. Klin. Wochenschr. 53, 1061–1067 (1975)
Meyer zum Büschenfelde, K.-H., Arnold, W., Knolle, J., Hess, G.: Immunreaktionen gegenüber HBsAG, HBcAG und e-Antigen bei akuter Virushepatitis sowie lebergesunden und leberkranken HBsAG-Trägern. Z. Gastroenterol. 14, 365 (1976)
Nayak, N.C., Sachdeva, R.: Localisation of hepatitis B surface antigen in conventional paraffin sections of liver. Am. J. Path. 81, 479 (1975)
Peters, R.L.: Viral hepatitis: a pathologic spectrum. Am. Med. Sci. 270, 17–31 (1975)
Portmann, B., Galbraith, R.M., Eddleston, A.L.W.F., Zuckermann, A.J., Williams, R.: Detection of HBsAG in fixed liver tissue — use of a modified immunofluorescent technique and comparison with histochemical methods. Gut 17, 1–9 (1976)
Poley, J.R.: Editorial: Virushepatitis — Fortschritt und Ausblick. Helv. Paediatr. Acta 32, 5–9 (1977)
Ray, M.B., Desmet, V.J.: Distribution patterns of hepatitis B surface antigen (HBsAG) in liver biopsies of hepatitis B patients. Acta Gastro-Enterol. Belg. 39, 307–317 (1976)
Ray, M.B., Desmet, V.J., Bradburne, A.F., Desmyter, J., Fevery, J., De Groote, J.: Differential distribution of hepatitis B surface antigen and hepatitis core antigen in the liver of hepatitis B patients. Gastroenterology 71, 462–469 (1976)
Robinson, W.S., Lutwick, L.I.: The virus of hepatitis, type B. N. Engl. J. Med. 295, 1168–1175 u. 1232–1236 (1976)
Salaspuro, M., Sipponen, P.: Demonstration of an intracellular copper-binding protein by orcein staining in long standing cholestatic liver diseases. Gut 17, 787–790 (1976)
Sakurai, M., Mikyaji, T.: Orcein staining of hepatitis B surface antigen in paraffin sections of liver in autopsy cases. Acta Hepato-Gastroenterol. 24, 334–339 (1977)
Shikata, T.: Australia-Antigen in liver tissue — An immunoelectron microscopy study. Jap. J. Exp. Med. 43, 231–245 (1973)
Shikata, T., Uzawa, T., Yoshiwara, N., Akatsuko, T., Yamazaki, S.: Staining methods of Australia antigen in paraffin section. Detection of cytoplasmic inclusion bodies. Jap. J. Exp. Med. 44, 25–36 (1974)
Scheuer, P.J.: Primary biliary cirrhosis. Proc. R. Soc. Med. 60, 1257–1260 (1967)
Sipponen, P.: Orcein positive hepatocellular material in long-standing biliary diseases. I. Histochemical characteristics. Scand. J. Gastroenterol. 11, 545–552 (1976a)
Sipponen, P.: Orcein positive hepatocellular material in long-standing biliary diseases. II. Ultrastructural studies. Scand. J. Gastroenterol. 11, 553–557 (1976b)
Sumithran, E.: Methods for detection of hepatitis B surface antigen in paraffin sections of liver: a guideline for their use. J. Clin. Path. 30, 460–463 (1976)
Sumithran, E., Prathap, K.: HBsAG-positive chronic liver disease associated with cirrhosis and hepatocellular carcinoma in the Senoi, Cancer 40, 1618–1620 (1977)
Thomsen, P., Poulsen, H., Petersen, P.: Different types of ground-glass hepatocytes in human liver biopsies: Morphology, occurrence and diagnostic significance. Scand. J. Gastroenterol. 11, 113–119 (1976)
Turbitt, M.L., Patrick, R.S., Goudie, R.B., Buchanan, W.M.: Incidence in south-west Scotland of hepatitis surface antigen in the liver of patients with hepatocellular carcinoma. J. Clin. Path. 30, 1124–1128 (1977)
Vogel, H.M., Henning, H., Lüders, C.J., Tripatzis, I., v. Braun, H.H.: Lichtoptischer Nachweis von HBsAG-Befunden im Leberbioptat — Vergleichende Untersuchungen von HB-AG-Befunden im Serum mit Ergebnissen einer Aldehydthionin-Färbung. Gastroenterology 12, 257–264 (1974)
Winckler, K., Junge, U., Creutzfeldt, W.: Ground-glass hepatocytes in unselected liver biopsies. Ultrastructure and relationship to hepatitis B surface antigen. Scand. J. Gastroenterol. 11, 167–170 (1976)
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Herrn Professor Dr. Drs. h.c. H. Meessen zum 70. Geburtstag gewidmet
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Borchard, F., Gussmann, V. Detection of HBsAg containing cells in liver biopsies by different stains and classification of positively reacting ground-glass hepatocytes. Virchows Arch. A Path. Anat. and Histol. 384, 245–261 (1979). https://doi.org/10.1007/BF00428227
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DOI: https://doi.org/10.1007/BF00428227