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. 1982 Sep;49(3):543–551.

Characterization of antigen moiety of HBsAg in the complement fixing immune complexes of hepatitis B virus positive chronic active hepatitis.

R C Gupta
PMCID: PMC1536728  PMID: 7172494

Abstract

Circulating immune complexes (CIC) are frequently found in hepatitis B virus-induced chronic active hepatitis. Since antigen and antibody moieties of complexes are critical in determining many of its pathogenic factors, the constituents of these complexes were investigated with particular attention to the quantity and nature of the HBs antigen moiety of the complexes. Complement fixing immune complexes were isolated from sera of 14 patients with chronic active hepatitis by utilizing conglutinin's unique property to bind C3-fixed complexes. Low pH (2.6) was used to dissociate the complexes. Components of dissociated complexes were separated into antigen and antibody fractions using immobilized Protein A. Both fractions were analysed by electrophoresis in polyacrylamide gel with SDS. The antibody fractions showed heavy and light chains of IgG and IgM. The antigen fractions demonstrated six to 10 protein bands with mol. wt ranging between 17,000 and 120,000 daltons. To define precisely the polypeptide antigen moiety involved in the immune complex formation, a transfer blotting technique was used employing human anti-HBs globulin as probe. Polypeptides with mol. wt 97,000 and 49,000 reacted as antigen moieties of HBsAg. In addition, the levels of HBsAg in the antigen fractions were significantly greater (P less than 0.005) compared to sera from patients with acute hepatitis. Implications of these findings are discussed.

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Selected References

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

  1. Almeida J. D., Waterson A. P. Immune complexes in hepatitis. Lancet. 1969 Nov 8;2(7628):983–986. doi: 10.1016/s0140-6736(69)90540-6. [DOI] [PubMed] [Google Scholar]
  2. Carella G., Digeon M., Feldmann G., Jungers P., Drouet J., Bach J. F. Detection of hepatitis B antigen in circulating immune complexes in acute and chronic hepatitis. Scand J Immunol. 1977;6(12):1297–1304. doi: 10.1111/j.1365-3083.1977.tb00369.x. [DOI] [PubMed] [Google Scholar]
  3. Casali P., Bossus A., Carpentier N. A., Lambert P. H. Solid-phase enzyme immunoassay or radioimmunoassay for the detection of immune complexes based on their recognition by conglutinin: conglutinin-binding test. A comparative study with 125I-labelled Clq binding and Raji-cell RIA tests. Clin Exp Immunol. 1977 Aug;29(2):342–354. [PMC free article] [PubMed] [Google Scholar]
  4. Digeon M., Laver M., Riza J., Bach J. F. Detection of circulating immune complexes in human sera by simplified assays with polyethylene glycol. J Immunol Methods. 1977;16(2):165–183. doi: 10.1016/0022-1759(77)90051-5. [DOI] [PubMed] [Google Scholar]
  5. Gerber M. A., Sarno E., Vernace S. J. Immune complexes in hepatocytic nuclei of HB ag-positive chronic hepatitis. N Engl J Med. 1976 Apr 22;294(17):922–925. doi: 10.1056/NEJM197604222941704. [DOI] [PubMed] [Google Scholar]
  6. Gupta R. C., McDuffie F. C., Tappeiner G., Jordon R. E. Binding of soluble immune complexes to Raji lymphocytes. Role of receptors for complement components, C1q and C3-C3b. Immunology. 1978 Apr;34(4):751–761. [PMC free article] [PubMed] [Google Scholar]
  7. Gupta R. C., Tan E. M. Isolation of circulating immune complexes by conglutinin and separation of antigen from dissociated complexes by immobilized protein A. Clin Exp Immunol. 1981 Oct;46(1):9–19. [PMC free article] [PubMed] [Google Scholar]
  8. Lachmann P. J., Macanovic M., Harkiss G. D., Oldroyd R. G., Habicht J. The isolation of the antibody moieties of immune complexes from serum by the pepsin digestion of conglutinin-anti-conglutinin complexes. Clin Exp Immunol. 1981 Nov;46(2):250–258. [PMC free article] [PubMed] [Google Scholar]
  9. Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
  10. Mackay P., Burrell C. J. Examination of the polypeptides of hepatitis B surface antigen. J Gen Virol. 1976 Nov;33(2):181–191. doi: 10.1099/0022-1317-33-2-181. [DOI] [PubMed] [Google Scholar]
  11. Madaliński K., Bragiel I. HBsAg immune complexes in the course of infection with hepatitis B virus. Clin Exp Immunol. 1979 Jun;36(3):371–378. [PMC free article] [PubMed] [Google Scholar]
  12. Nowoslawski A., Krawczynski K., Nazarewicz T., Slusarczyk J. Immunopathological aspects of hepatitis type B. Am J Med Sci. 1975 Sep-Oct;270(2):229–239. doi: 10.1097/00000441-197509000-00002. [DOI] [PubMed] [Google Scholar]
  13. Nydegger U. E., Lambert P. H., Gerber H., Miescher P. A. Circulating immune complexes in the serum in systemic lupus erythematosus and in carriers of hepatitis B antigen. Quantitation by binding to radiolabeled C1q. J Clin Invest. 1974 Aug;54(2):297–309. doi: 10.1172/JCI107765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Pernice W., Sodomann C. P., Lüben G., Seiler F. R., Sedlacek H. H. Antigen-specific detection of HBsAG-containing immune complexes in the course of hepatitis B virus infection. Clin Exp Immunol. 1979 Aug;37(2):376–380. [PMC free article] [PubMed] [Google Scholar]
  15. Shih J. W., Gerin J. L. Proteins of hepatitis B surface antigen. J Virol. 1977 Jan;21(1):347–357. doi: 10.1128/jvi.21.1.347-357.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Shih J. W., Tan P. L., Gerin J. L. Antigenicity of the major polypeptides of hepatitis B surface antigen (HBsAg). J Immunol. 1978 Feb;120(2):520–525. [PubMed] [Google Scholar]
  17. Skelly J., Howard C. R., Zuckerman A. J. Analysis of hepatitis B surface antigen components solubilized with Triton X-100. J Gen Virol. 1979 Sep;44(3):679–689. doi: 10.1099/0022-1317-44-3-679. [DOI] [PubMed] [Google Scholar]
  18. Soloway R. D., Summerskill W. H., Baggenstoss A. H., Geall M. G., Gitnićk G. L., Elveback I. R., Schoenfield L. J. Clinical, biochemical, and histological remission of severe chronic active liver disease: a controlled study of treatments and early prognosis. Gastroenterology. 1972 Nov;63(5):820–833. [PubMed] [Google Scholar]
  19. Thomas H. C., De Villiers D., Potter B., Hodgson H., Jain S., Jewell D. P., Sherlock S. Immune complexes in acute and chronic liver disease. Clin Exp Immunol. 1978 Feb;31(2):150–157. [PMC free article] [PubMed] [Google Scholar]
  20. Thomas H. C., Potter B. J., Elias E., Sherlock S. Metabolism of the third component of complement in acute type B hepatitis, HBs antigen positive glomerulonephritis, polyarteritis nodosum, and HBs antigen positive and negative chronic active liver disease. Gastroenterology. 1979 Apr;76(4):673–679. [PubMed] [Google Scholar]
  21. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  23. Zacharius R. M., Zell T. E., Morrison J. H., Woodlock J. J. Glycoprotein staining following electrophoresis on acrylamide gels. Anal Biochem. 1969 Jul;30(1):148–152. doi: 10.1016/0003-2697(69)90383-2. [DOI] [PubMed] [Google Scholar]

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