Production of infectious hepatitis C virus particles in three-dimensional cultures of the cell line carrying the genome-length dicistronic viral RNA of genotype 1b.
We show that a dicistronic hepatitis C virus (HCV) genome of genotype 1b supports the production and secretion of infectious HCV particles in two independent three-dimensional (3D) culture systems, the radial-flow bioreactor and the thermoreversible gelation polymer (TGP), but not in monolayer cultures. Immunoreactive enveloped particles, which are 50-60 nm in diameter and are surrounded by membrane-like structures, are observed in the culture medium as well as at the endoplasmic reticulum membranes and in dilated cytoplasmic cisternae in spheroids of Huh-7 cells. Infection of HCV particles is neutralized by anti-E2 antibody or patient sera that interfere with E2 binding to human cells. Finally, the utility of the 3D-TGP culture system for the evaluation of antiviral drugs is shown. We conclude that the replicon-based 3D culture system allows the production of infectious HCV particles. This system is a valuable tool in studies of HCV morphogenesis in a natural host cell environment.
Murakami
Kyoko
K
Department of Virology II, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan.
Ishii
Koji
K
Ishihara
Yousuke
Y
Yoshizaki
Sayaka
S
Tanaka
Keiko
K
Gotoh
Yasufumi
Y
Aizaki
Hideki
H
Kohara
Michinori
M
Yoshioka
Hiroshi
H
Mori
Yuichi
Y
Manabe
Noboru
N
Shoji
Ikuo
I
Sata
Tetsutaro
T
Bartenschlager
Ralf
R
Matsuura
Yoshiharu
Y
Miyamura
Tatsuo
T
Suzuki
Tetsuro
T