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Virology; Hepatitis C virus subgenomic replicons can be grown in nonhepatic cells

Posted on: Thursday, 12 February 2004, 06:00 CST

2004 FEB 16 - (NewsRx.com & NewsRx.net) -- Hepatitis C virus subgenomic replicons can be grown in nonhepatic cells.

"Hepatitis C virus (HCV) infects liver cells and its replication in other cells is incompletely defined. Human hepatoma Huh-7 cells harboring subgenomic HCV replicons were used in somatic cell fusion experiments with human embryonic kidney 293 cells as a means of examining the permissiveness of 293 cells for HCV subgenomic RNA replication. 293 cells were generally not permissive for replication of Huh-7 cell-adapted replicons," investigators in Canada report.

"However, upon coculturing of the two cell lines, we selected rare replicon-containing cells, termed 293Rep cells, that resembled parental 293 cells. Direct metabolic labeling of cells with P-33 in the presence of actinomycin D and Northern blotting to detect the negative strand of the replicon demonstrated functional RNA replicons in 293Rep cells," wrote S. Ali and colleagues, Boehringer Ingelheim Canada Ltd, Research & Development.

"Furthermore, Western blots revealed that 293Rep cells expressed the HCV nonstructural proteins as well as markers of the naive 293 cells but not Huh-7 cells. Propidium iodide staining and fluorescence-activated cell sorting analysis of 293Rep cells revealed that clone 293Rep17 closely resembled naive 293 cells."

"Transfection of total RNA from 293Rep17 into naive 293 cells produced replicon-containing 293 cell lines with characteristics distinct from those of Huh-7-derived replicon cell lines. Relative to Huh-7 replicons, the 293 cell replicons were less sensitive to inhibition by alpha interferon and substantially more sensitive to inhibition by poly(I)-poly(C) double-stranded RNA," study authors indicated.

"This study established HCV subgenomic replicons in nonhepatic 293 cells and demonstrated their utility in expanding the study of cellular HCV RNA replication," scientists concluded.

Ali and colleagues published their study in Journal of Virology (Hepatitis C virus subgenomic replicons in the human embryonic kidney 293 cell line. J Virol, 2004;78(1):491-501).

For additional information, contact G. Kukolj, Boehringer Ingelheim Canada Ltd, Research & Development, Department Biology Science, 2100 Rue Cunard, Laval, PQ H7S 2G5, Canada.

The publisher of the Journal of Virology can be contacted at: American Society for Microbiology, 1752 N St. NW, Washington, DC 20036-2904, USA.

The information in this article comes under the major subject areas of Hepatitis C Virus, Hepatology, Genomics and Genetics, and Virology.

This article was prepared by Biotech Business Week editors from staff and other reports. Copyright 2004, Biotech Business Week via NewsRx.com & NewsRx.net.

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