Detection of hepatitis B surface antigen (HBsAg) in peripheral blood mononuclear cells of patients with acute and chronic active hepatitis B

Abstract

Seventy five patients with acute and chronic active hepatitis (CAH) were studied by indirect immunofluorescence with monoclonal antibodies for the presence of hepatitis B surface antigen (HBsAg) on peripheral blood mononuclear cells (PBMC). The viral surface antigen was detected in the PBMC of all the patients with hepatitis B virus (HBV)-induced CAH and in acute patients with more than 2 months of evolution. No HBsAg was detected in the samples obtained from 12 normal controls or from 14 non-A, non-B CAH patients. Analysis of PBMC subsets revealed that HBsAg was present in non-T cells; dual fluorescence studies showed HBsAg on surface Ig-positive lymphocytes. The binding of anti-HBs monoclonal antibodies was higher than that of a goat anti-HBs serum, and the highest reactivity was observed with an antibody against the pre-S(2)-region sequence. Both HBsAg and hepatitis B core antigen (HBcAg) were also detected in lysates of PBMC by dot blot analysis.

This is a preview of subscription content, access via your institution.

References

  1. 1.

    Gerber MA, Thung SN: Molecular and cellular pathology of hepatitis B. Lab Invest 52:572–588, 1985.

    Google Scholar 

  2. 2.

    Dienstang JL: Immunologic mechanisms in chronic viral hepatitis.In Viral Hepatitis and Liver Disease, GN Vyas, JL Dienstang, JH Hoofnagle (eds). Orlando, FL, Grune & Stratton, 1984, pp 135–166

    Google Scholar 

  3. 3.

    Blum HE, Stowring L, Figus A, Montgomery CK, Haase AT, Vyas GN: Detection of hepatitis B virus DNA in hepatocytes, bile duct epithelium, and vascular elements by in situ hybridization. Proc Natl Acad Sci USA 88:6685–6688, 1983

    Google Scholar 

  4. 4.

    Siddiqui A: Hepatitis B virus DNA in Kaposi sarcoma. Proc Natl Acad Sci USA 79:5675–5679, 1983

    Google Scholar 

  5. 5.

    Dejean A, Lugassy C, Zafrani S, Tiollais D, Brechot C: Detection of hepatitis B virus DNA in pancreas, kidney and skin of two human carriers of the virus. J Gen Virol 65:651–655, 1984

    Google Scholar 

  6. 6.

    Romet-Lemonne JL, McLane MT, Elfassi E, Haseltine WA, Azocar J, Essex M: Hepatitis B virus infection in cultured human lymphoblastoid cells. Science 221:667–669, 1983

    Google Scholar 

  7. 7.

    Elfassi E, Romet-Lemonne JL, Essex M, McLane F, Haseltine WA: Evidence of extrachromosomal forms of hepatitis B viral DNA in a bone marrow culture obtained from a patient recently infected with hepatitis B virus. Proc Natl Acad Sci USA 81:3526–3528, 1984

    Google Scholar 

  8. 8.

    Pontisso P, Doon MC, Tiollais P, Brechot C: Detection of hepatitis B virus DNA in mononuclear blood cells. Br Med J 288:1563–1566, 1984

    Google Scholar 

  9. 9.

    Lee-Injo LE, Balasegaram M, Lopez CG, Herrera AR: Hepatitis B virus DNA in liver and white blood cells of patients with hepatoma. DNA 2:301–308, 1983

    Google Scholar 

  10. 10.

    Pasquinelli C, Laure F, Chatenoud L, Beaurin G, Gazengel C, Bismuth H, Degos F, Tiollais P, Bach JF, Brechot C: Hepatitis B virus DNA in mononuclear blood cells. A frequent event in hepatitis B surface antigen-positive and -negative patients with acute and chronic liver disease. J Hepatol 3:95–103, 1986

    Google Scholar 

  11. 11.

    Yoffe B, Noonan CA, Melnick JL, Hollinger FB: Hepatitis B virus DNA in mononuclear cells and analysis of cell subsets for the presence of replicative intermediates of viral DNA. J Infect Dis 153:471–477, 1986

    Google Scholar 

  12. 12.

    Noonan CA, Yoffe B, Mansell PWA, Melnick JL, Hollinger FB: Extrachromosomal sequences of hepatitis B virus DNA in peripheral blood mononuclear cells of acquired immune deficiency syndrome patients. Proc Natl Acad Sci USA 83:5698–5702, 1986

    Google Scholar 

  13. 13.

    Korba BE, Wells F, Tennant BC, Yoakun GH, Purcell RH, Gerin JL: Hepadnavirus infection of peripheral blood lymphocytes in vivo: Woodchuck and chimpanzee models of viral hepatitis. J Virol 58:1–8, 1986

    Google Scholar 

  14. 14.

    Boyum A: Separation of blood leukocytes, granulocytes and lymphocytes. Tissue Antigens 4:269–273, 1974

    Google Scholar 

  15. 15.

    Bodmer JG, Pickbourne P, Richards S: Ia serology.In Histocompatibility Testing 1977, WF Bodmeret al. (eds). Copenhagen, Munksgaard, 1978, pp 35–84

    Google Scholar 

  16. 16.

    Roseto A, Scherrer R, Cohen J, Guillemin MC, Charlipienne A, Feynerol C, Peries J: Isolation and characterization of anti-rotavirus immunoglobulins secreted by cloned hybridoma cell lines. J Gen Virol 64:237–241, 1983

    Google Scholar 

  17. 17.

    Hess S, Shih JW, Arnold W: Demonstration and partial characterization of 22 nm HBsAg and Dane particles of subtype HBsAg/ady. J Immunol 123:1189–1193, 1979

    Google Scholar 

  18. 18.

    Herbrink P, Van Bussel FJ, Warnaar SO: The antigen spot test (AST): A highly sensitive assay for the detection of antibodies. J Immunol Meth 48:293–298, 1982

    Google Scholar 

  19. 19.

    De Blas AL, Cherwinski HM: Detection of antigens on nitrocellulose paper immunoblots with monoclonal antibodies. Anal Biochem 133:214–219, 1983

    Google Scholar 

  20. 20.

    Fridman WH, Gresser, Bandu MT, Auget M, Neauport-Sautes C: Interferon enhances expression of Fc receptors. J Immunol 124:2436–2441, 1980

    Google Scholar 

  21. 21.

    Auget M, Vignaux F, Fridman WH, Gresser I: Enhancement of Fc receptor expression in interferon-treated mice. Eur J Immunol 11:926–930, 1981

    Google Scholar 

Download references

Author information

Affiliations

Authors

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Roisman, F.R., Mota, A.H. & Fainboim, L. Detection of hepatitis B surface antigen (HBsAg) in peripheral blood mononuclear cells of patients with acute and chronic active hepatitis B. J Clin Immunol 9, 10–15 (1989). https://doi.org/10.1007/BF00917122

Download citation

Key words

  • Hepatitis B virus
  • hepatitis B surface antigen (HBsAg) detection
  • chronic active hepatitis