Southern-blot analysis and simultaneous in situ detection of hepatitis B virus-associated DNA and antigens in patients with end-stage liver disease

Kwang Hyub Han, F. Blaine Hollinger, Christine A. Noonan, Harvey Solomon, Goran B G Klintmalm, Robert M. Genta, Boris Yoffe

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Abstract

To gain new insights into the pathogenesis of hepatitis B virus-induced chronic liver disease, we have used nonisotopic in situ detection methods for the simultaneous analysis of hepatitis B virus DNA and antigens at the single-cell level. Paraffin-embedded liver specimens from 23 cirrhotic patients (12 HBsAg positive and 11 HBsAg negative) who underwent liver transplantation were evaluated by in situ hybridization with a digoxigenin-labeled DNA probe and digoxigenin detection system and by immunohistochemistry with an enhanced biotin-streptavidin technique. DNAs extracted from liver and serum specimens were analyzed by Southern- and slot-blot hybridization, respectively. Using the in situ techniques, we detected hepatitis B virus-specific DNA and antigens in 11 of 12 HBsAg-positive patients and in none of the 11 HBsAg-negative individuals. Replicative intermediates of hepatitis B virus DNA were detected by Southern-blot analysis in the same 11 HBsAg-positive patients, 6 of whom had no serological markers of hepatitis B virus replication. Therefore a good correlation was found between the results obtained by the in situ and Southern-blot hybridization analyses of tissue specimens. However, a lack of correlation was found between serum- and tissue-associated markers of viral replication. In addition, the simultaneous in situ detection analyses revealed that some hepatocytes containing high levels of viral DNA were devoid of detectable HBcAg, suggesting a mechanism by which the virus may escape immunological surveillance. These data provide evidence that liver-associated HBV replication may persist in the absence of serological markers of active hepatitis B virus replication in cirrhotic patients with advanced liver disease and demonstrate that the evaluation of liver- rather than serum-associated markers of viral replication provides a more accurate assessment of the virological events occurring in HBsAg-positive individuals.

Original languageEnglish (US)
Pages (from-to)1032-1038
Number of pages7
JournalHepatology
Volume18
Issue number5
StatePublished - Nov 1993

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End Stage Liver Disease
Hepatitis B Surface Antigens
Southern Blotting
Hepatitis B virus
Antigens
DNA
Digoxigenin
Liver
Virus Replication
Liver Diseases
Immunologic Surveillance
Biomarkers
Hepatitis B Antigens
Hepatitis B Core Antigens
Streptavidin
DNA Probes
Viral DNA
Biotin
Serum
Liver Transplantation

ASJC Scopus subject areas

  • Hepatology

Cite this

Han, K. H., Hollinger, F. B., Noonan, C. A., Solomon, H., Klintmalm, G. B. G., Genta, R. M., & Yoffe, B. (1993). Southern-blot analysis and simultaneous in situ detection of hepatitis B virus-associated DNA and antigens in patients with end-stage liver disease. Hepatology, 18(5), 1032-1038.

Southern-blot analysis and simultaneous in situ detection of hepatitis B virus-associated DNA and antigens in patients with end-stage liver disease. / Han, Kwang Hyub; Hollinger, F. Blaine; Noonan, Christine A.; Solomon, Harvey; Klintmalm, Goran B G; Genta, Robert M.; Yoffe, Boris.

In: Hepatology, Vol. 18, No. 5, 11.1993, p. 1032-1038.

Research output: Contribution to journalArticle

Han, KH, Hollinger, FB, Noonan, CA, Solomon, H, Klintmalm, GBG, Genta, RM & Yoffe, B 1993, 'Southern-blot analysis and simultaneous in situ detection of hepatitis B virus-associated DNA and antigens in patients with end-stage liver disease', Hepatology, vol. 18, no. 5, pp. 1032-1038.
Han, Kwang Hyub ; Hollinger, F. Blaine ; Noonan, Christine A. ; Solomon, Harvey ; Klintmalm, Goran B G ; Genta, Robert M. ; Yoffe, Boris. / Southern-blot analysis and simultaneous in situ detection of hepatitis B virus-associated DNA and antigens in patients with end-stage liver disease. In: Hepatology. 1993 ; Vol. 18, No. 5. pp. 1032-1038.
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