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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1984 May;81(10):3195–3199. doi: 10.1073/pnas.81.10.3195

Detection of non-A, non-B hepatitis antigen by immunocytochemical staining.

K H Burk, P E Oefinger, G R Dreesman
PMCID: PMC345248  PMID: 6203116

Abstract

Liver tissue obtained from a chimpanzee during the acute phase of an experimental non-A, non-B (NANB) hepatitis virus infection was studied by a sensitive immunocytochemical staining procedure for the presence of NANB viral antigens. Initial investigations were conducted with a model system of hepatitis B virus (HBV) antigens for purposes of comparing two immunocytochemical staining methods. Of these two procedures, an immunoperoxidase procedure, utilizing an avidin-biotinylated enzyme complex, was at least 40-fold more sensitive than a conventional immunoperoxidase technique for the detection of HBV-specific tissue antigens. Utilization of the avidin-biotin-amplified immunoperoxidase staining procedure, in conjunction with four primary convalescent antisera obtained from NANB hepatitis-implicated donors, resulted in the observation of NANB virus-associated antigen in the cytoplasm of hepatocytes from an infected chimpanzee liver. These same human antisera were not reactive with a number of uninfected control cells nor with cells infected with HBV, hepatitis A virus, or cytomegalovirus. Preincubation of one of these convalescent NANB sera, or IgG derived thereof, with an acute-phase serum obtained from a NANB hepatitis virus-infected chimpanzee abolished the antibody reactivity. We conclude from these observations that selected convalescent sera from NANB hepatitis virus-infected patients contain low levels of antibody that specifically react with a cytoplasmic antigen associated with NANB virus-infected hepatocytes.

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Selected References

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  1. Alter H. J., Holland P. V., Morrow A. G., Purcell R. H., Feinstone S. M., Moritsugu Y. Clinical and serological analysis of transfusion-associated hepatitis. Lancet. 1975 Nov 1;2(7940):838–841. doi: 10.1016/s0140-6736(75)90234-2. [DOI] [PubMed] [Google Scholar]
  2. Alter H. J., Purcell R. H., Holland P. V., Popper H. Transmissible agent in non-A, non-B hepatitis. Lancet. 1978 Mar 4;1(8062):459–463. doi: 10.1016/s0140-6736(78)90131-9. [DOI] [PubMed] [Google Scholar]
  3. Berman M., Alter H. J., Ishak K. G., Purcell R. H., Jones E. A. The chronic sequelae of non-A, non-B hepatitis. Ann Intern Med. 1979 Jul;91(1):1–6. doi: 10.7326/0003-4819-91-1-1. [DOI] [PubMed] [Google Scholar]
  4. Bradley D. W., Cook E. H., Maynard J. E., McCaustland K. A., Ebert J. W., Dolana G. H., Petzel R. A., Kantor R. J., Heilbrunn A., Fields H. A. Experimental infection of chimpanzees with antihemophilic (factor VIII) materials: recovery of virus-like particles associated with non-A, non-B hepatitis. J Med Virol. 1979;3(4):253–269. doi: 10.1002/jmv.1890030403. [DOI] [PubMed] [Google Scholar]
  5. Bradley D. W., Maynard J. E., Popper H., Ebert J. W., Cook E. H., Fields H. A., Kemler B. J. Persistent non-A, non-B hepatitis in experimentally infected chimpanzees. J Infect Dis. 1981 Feb;143(2):210–218. doi: 10.1093/infdis/143.2.210. [DOI] [PubMed] [Google Scholar]
  6. Burk K. H., Cabral G. A., Dreesman G. R., Peters R. L., Alter H. J. Ultrastructural changes and virus-like particles localized in liver hepatocytes of chimpanzees infected with non-A, non-B hepatitis. J Med Virol. 1981;7(1):1–19. doi: 10.1002/jmv.1890070102. [DOI] [PubMed] [Google Scholar]
  7. Cabral G. A., Gyorkey F., Gyorkey P., Melnick J. L., Dreesman G. R. Immunohistochemical and electron microscopic detection of hepatitis B surface and core antigens. Exp Mol Pathol. 1978 Oct;29(2):156–169. doi: 10.1016/0014-4800(78)90036-9. [DOI] [PubMed] [Google Scholar]
  8. Chircu L. V., Pezzella M., Lacava V., Ricci G. Post-transfusion hepatitis: antigen/antibody systems correlated with non-A, non-B hepatitis. J Med Virol. 1980;6(2):147–151. doi: 10.1002/jmv.1890060207. [DOI] [PubMed] [Google Scholar]
  9. Duermeyer W., Stute R., Hellings J. A. An enzyme-linked immunosorbent assay for an antigen related to non-A, non-B hepatitis and its antibody: partial characterization of the antigen and chimpanzee transmission. J Med Virol. 1983;11(1):11–21. doi: 10.1002/jmv.1890110103. [DOI] [PubMed] [Google Scholar]
  10. Hollinger F. B., Gitnick G. L., Aach R. D., Szmuness W., Mosley J. W., Stevens C. E., Peters R. L., Weiner J. M., Werch J. B., Lander J. J. Non-A, non-B hepatitis transmission in chimpanzees: a project of the transfusion-transmitted viruses study group. Intervirology. 1978;10(1):60–68. doi: 10.1159/000148969. [DOI] [PubMed] [Google Scholar]
  11. Hoofnagle J. H., Gerety R. J., Tabor E., Feinstone S. M., Barker L. F., Purcell R. H. Transmission of non-A, non-B hepatitis. Ann Intern Med. 1977 Jul;87(1):14–20. doi: 10.7326/0003-4819-87-1-14. [DOI] [PubMed] [Google Scholar]
  12. Hoofnagle J. H. Precipitin system detected in sera from patients with non-A, non-B hepatitis. J Med Virol. 1981;7(4):315–319. doi: 10.1002/jmv.1890070407. [DOI] [PubMed] [Google Scholar]
  13. Hsu S. M., Raine L., Fanger H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem. 1981 Apr;29(4):577–580. doi: 10.1177/29.4.6166661. [DOI] [PubMed] [Google Scholar]
  14. Iwarson S., Lindberg J., Lundin P. Progression of hepatitis non-A, non-B to chronic active hepatitis: a histological follow-up of two cases. J Clin Pathol. 1979 Apr;32(4):351–355. doi: 10.1136/jcp.32.4.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kabiri M., Tabor E., Gerety R. J. Antigen-antibody system associated with non-A, non-B hepatitis detected by indirect immunofluorescence. Lancet. 1979 Aug 4;2(8136):221–224. doi: 10.1016/s0140-6736(79)90237-x. [DOI] [PubMed] [Google Scholar]
  16. Kendall C., Ionescu-Matiu I., Dreesman G. R. Utilization of the biotin/avidin system to amplify the sensitivity of the enzyme-linked immunosorbent assay (ELISA). J Immunol Methods. 1983 Feb 11;56(3):329–339. doi: 10.1016/s0022-1759(83)80022-2. [DOI] [PubMed] [Google Scholar]
  17. Kim H. C., Saidi P., Ackley A. M., Bringelsen K. A., Gocke D. J. Prevalence of type B and non-A, non-B hepatitis in hemophilia: relationship to chronic liver disease. Gastroenterology. 1980 Dec;79(6):1159–1164. [PubMed] [Google Scholar]
  18. Mounts P., Shah K. V., Kashima H. Viral etiology of juvenile- and adult-onset squamous papilloma of the larynx. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5425–5429. doi: 10.1073/pnas.79.17.5425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nerurkar L. S., Jacob A. J., Madden D. L., Sever J. L. Detection of genital herpes simplex infections by a tissue culture-fluorescent-antibody technique with biotin-avidin. J Clin Microbiol. 1983 Jan;17(1):149–154. doi: 10.1128/jcm.17.1.149-154.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Neurath A. R., Stevens C. E., Strick N., Szmuness W., Oleszko W. R., Harley E. J. An antigen detected frequently in human sera with elevated levels of alanine aminotransferase: a potential marker for non-A, non-B hepatitis. J Gen Virol. 1980 Jun;48(Pt 2):285–295. doi: 10.1099/0022-1317-48-2-285. [DOI] [PubMed] [Google Scholar]
  21. Notani G. W., Parsons J. A., Erlandsen S. L. Versatility of Staphylococcus aureus protein A in immunocytochemistry. Use in unlabeled antibody enzyme system and fluorescent methods. J Histochem Cytochem. 1979 Nov;27(11):1438–1444. doi: 10.1177/27.11.390040. [DOI] [PubMed] [Google Scholar]
  22. Oefinger P. E., Bronson D. L., Dreesman G. R. Induction of hepatitis B surface antigen in human hepatoma-derived cell lines. J Gen Virol. 1981 Mar;53(Pt 1):105–113. doi: 10.1099/0022-1317-53-1-105. [DOI] [PubMed] [Google Scholar]
  23. Prince A. M. Non-A, non-B hepatitis viruses. Annu Rev Microbiol. 1983;37:217–232. doi: 10.1146/annurev.mi.37.100183.001245. [DOI] [PubMed] [Google Scholar]
  24. Seeff L. B., Wright E. C., Zimmerman H. J., McCollum R. W. Va cooperative study of post-transfusion hepatitis, 1969-1974: incidence and characteristics of hepatitis and responsible risk factors. Am J Med Sci. 1975 Sep-Oct;270(2):355–362. doi: 10.1097/00000441-197509000-00018. [DOI] [PubMed] [Google Scholar]
  25. Shimizu Y. K., Feinstone S. M., Purcell R. H., Alter H. J., London W. T. Non-A, non-B hepatitis: ultrastructural evidence for two agents in experimentally infected chimpanzees. Science. 1979 Jul 13;205(4402):197–200. doi: 10.1126/science.451589. [DOI] [PubMed] [Google Scholar]
  26. Shirachi R., Shiraishi H., Tateda A., Kikuchi K., Ishida N. Hepatitis "C" antigen in non-A, non-B post-transfusion hepatitis. Lancet. 1978 Oct 21;2(8095):853–856. doi: 10.1016/s0140-6736(78)91567-2. [DOI] [PubMed] [Google Scholar]
  27. Suh D. J., White Y., Eddleston A. L., Amini S., Tsiquaye K., Zuckerman A. J., Williams R. Specificity of an immunoprecipitin test for non-A non-B hepatitis. Lancet. 1981 Jan 24;1(8213):178–180. doi: 10.1016/s0140-6736(81)90059-3. [DOI] [PubMed] [Google Scholar]
  28. Tabor E., Gerety R. J., Drucker J. A., Seeff L. B., Hoofnagle J. H., Jackson D. R., April M., Barker L. F., Pineda-Tamondong G. Transmission of non-A, non-B hepatitis from man to chimpanzee. Lancet. 1978 Mar 4;1(8062):463–466. doi: 10.1016/s0140-6736(78)90132-0. [DOI] [PubMed] [Google Scholar]
  29. Tabor E., Gerety R. J. Non-A, non-B hepatitis: new findings and prospects for prevention. Transfusion. 1979 Nov-Dec;19(6):669–674. doi: 10.1046/j.1537-2995.1979.19680104091.x. [DOI] [PubMed] [Google Scholar]
  30. Tabor E., Mitchell F. D., Goudeau A. M., Gerety R. J. Detection of an antigen-antibody system in serum associated with human non-A, non-B hepatitis. J Med Virol. 1979;4(3):161–169. doi: 10.1002/jmv.1890040302. [DOI] [PubMed] [Google Scholar]
  31. Tabor E., Seeff L. B., Gerety R. J. Chronic non-A, non-B hepatitis carrier state: transmissible agent documented in one patient over a six-year period. N Engl J Med. 1980 Jul 17;303(3):140–143. doi: 10.1056/NEJM198007173030307. [DOI] [PubMed] [Google Scholar]
  32. Trepo C., Vitvitski L., Hantz O., Chevallier P., Pichoud C., Babin S., Grimaud J. A., Sepetjan M. Identification and detection of long incubation non-A, non-B hepatitis virus and associated antigens or antibodies. J Virol Methods. 1980 Dec;2(1-2):127–139. doi: 10.1016/0166-0934(80)90046-4. [DOI] [PubMed] [Google Scholar]
  33. Tsiquaye K. N., Bird R. G., Tovey G., Wyke R. J., Williams R., Zuckerman A. J. Further evidence of cellular changes associated with non-A, non-B hepatitis. J Med Virol. 1980;5(1):63–71. doi: 10.1002/jmv.1890050108. [DOI] [PubMed] [Google Scholar]
  34. Vitvitski L., Trepo C., Prince A. M., Brotman B. Detection of virus-associated antigen in serum and liver of patients with non-A non-B hepatitis. Lancet. 1979 Dec 15;2(8155):1263–1267. doi: 10.1016/s0140-6736(79)92280-3. [DOI] [PubMed] [Google Scholar]

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