Transmission of HCV to a chimpanzee using virus particles produced in an RNA‐transfected HepG2 cell culture

Abstract

It was demonstrated previously that HepG2 cells produce negative strand RNA and virus-like particles after transfection with RNA transcribed from a full-length hepatitis C virus (HCV) cDNA clone [Dash et al. (1997) American Journal of Pathology, 151:363-373]. To determine in vivo infectivity of these in vitro synthesized viral particles, a chimpanzee was inoculated intravenously with HCV derived from HepG2 cells. The infected chimpanzee was examined serially for elevation of liver enzymes, for the presence of HCV RNA in the serum by reverse transcription nested polymerase chain reaction (RT-PCR), anti-HCV antibodies in the serum, and inflammation in the liver. The chimpanzee developed elevated levels of liver enzymes after the second week, but the levels fluctuated over a 10-week period. HCV RNA was detected in the serum of the chimpanzee at the second, seventh and ninth weeks after inoculation, and remained positive up to 25 weeks. Liver biopsies at Weeks 18 and 19 revealed of mild inflammation. Nucleotide sequence analysis of HCV recovered from the infected chimpanzee at the second and ninth weeks showed 100% sequence homology with the clone used for transfection studies. Serum anti-HCV antibodies were not detected by EIA during the 25 weeks follow-up period. These results suggest that intravenous administration of the virus-like particles derived from RNA-transfected HepG2 cells are infectious, and therefore, the pMO9.6-T7 clone is an infectious clone. These results provide new information that in vitro synthesized HCV particles produced from full-length HCV clone can cause infection in a chimpanzee. This study will facilitate the use of innovative approaches to the study of assembly of HCV particles and mechanisms of virus infectivity in cell culture.

RETRACTED ARTICLE: Small interfering RNA targeted to stem-loop II of the 5' untranslated region effectively inhibits expression of six HCV genotypes

“…We tested whether this highly effective siRNA-74 target in the 5' UTR region could silence the gene expression using full-length infectious clones of HCV1a and HCV1b. One HCV1a infectious clone (pCV-H77C) and two HCV1b infectious clones of HCV (pMO9.6-T7 and pCVJ4L6S) were used as targets [26-28]. To examine the antiviral effect of siRNA-74, each full-length clone were co-transfected with increasing concentration of siRNA plasmid using a two-step transfection procedure described earlier (37–38).…”

Hepatitis C virus targets over‐expression of arginase I in hepatocarcinogenesis

“…The clone used for this was a full-length, HCV cDNA of genotype 1b that was shown to replicate in cell culture (25) and was infectious in chimpanzees (26). This cDNA was previously sequenced, cloned into the mammalian expression vector pRc/CMV2, and then stably transfected into the human hepatoblastoma cell line, HepG2 and the human hepatoma cell line, Huh-7, as described (9).…”

Pathogenesis of Hepatitis C Virus Infection in Tupaia belangeri

“…The chimpanzee is the only validated animal model for in vivo studies of HCV infection, and it is capable of reproducing most aspects of human infection (5,18,23,28,35,36). The chimpanzee is also the only validated animal for testing the authenticity and infectivity of cloned viral sequences (8,14,35,36).…”