High-frequency transfection and cytopathology of the hepatitis B virus core antigen gene in human cells

George H. Yoakum, Brent E. Korba, John F. Lechner, Takayoshi Tokiwa, Adi F. Gazdar, Todd Seeley, Mary Siegel, Lawrence Leeman, Herman Autrup, Curtis C. Harris

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Abstract

A protoplast fusion method was developed to stably transfect human cells with pSV2-derived plasmids at frequencies greater than 10-3. This procedure made it possible to test the biological effect of a hepatitis B virus (HBV) gene independent of the viral structures required for infection. A pSV2gpt+ plasmid constructed to carry a subgenomic fragment of HBV that contained the core antigen gene (HBc gene) was transfected into human cells. A human epithelial cell line was stably transfected with the HBc + gene by selecting recipient cells for expression of guanine phosphoribosyl transferase expression. With this gpt+/HBc+ cell line it was shown that growth in serum-free medium or treatment with 5′-azacytidine stimulates the production of the HBV core antigen. A hepatocellular carcinoma carrying the entire HBV genome was stimulated to produce the HBc gene product in response to the same factors that stimulated HBcAg production in the gpt+/HBc+ cell line constructed by transfection. The temporal relation between the cytopathologic response and HBc gene expression was similar for both cell types, indicating a primary role for HBc gene expression in the cytopathology of HBV-infected human liver.

Original languageEnglish (US)
Pages (from-to)385-389
Number of pages5
JournalScience
Volume222
Issue number4622
DOIs
StatePublished - Jan 1 1983

ASJC Scopus subject areas

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    Yoakum, G. H., Korba, B. E., Lechner, J. F., Tokiwa, T., Gazdar, A. F., Seeley, T., Siegel, M., Leeman, L., Autrup, H., & Harris, C. C. (1983). High-frequency transfection and cytopathology of the hepatitis B virus core antigen gene in human cells. Science, 222(4622), 385-389. https://doi.org/10.1126/science.6194563