scholarly journals Characterization of a Cell Culture System of Persistent Hepatitis E Virus Infection in the Human HepaRG Hepatic Cell Line

Viruses ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 406
Author(s):  
Marie Pellerin ◽  
Edouard Hirchaud ◽  
Yannick Blanchard ◽  
Nicole Pavio ◽  
Virginie Doceul
Keyword(s):  
Cell Culture ◽  
Culture System ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Inhibitory Effect ◽  
Cell Culture System ◽  
Efficient Replication ◽  
A Cell ◽  
Vitro Model

Hepatitis E virus (HEV) is considered as an emerging global health problem. In most cases, hepatitis E is a self-limiting disease and the virus is cleared spontaneously without the need of antiviral therapy. However, immunocompromised individuals can develop chronic infection and liver fibrosis that can progress rapidly to cirrhosis and liver failure. The lack of efficient and relevant cell culture system and animal models has limited our understanding of the biology of HEV and the development of effective drugs for chronic cases. In the present study, we developed a model of persistent HEV infection in human hepatocytes in which HEV replicates efficiently. This HEV cell culture system is based on differentiated HepaRG cells infected with an isolate of HEV-3 derived from a patient suffering from acute hepatitis E. Efficient replication was maintained for several weeks to several months as well as after seven successive passages on HepaRG naïve cells. Moreover, after six passages onto HepaRG, we found that the virus was still infectious after oral inoculation into pigs. We also showed that ribavirin had an inhibitory effect on HEV replication in HepaRG. In conclusion, this system represents a relevant and efficient in vitro model of HEV replication that could be useful to study HEV biology and identify effective antiviral drugs against chronic HEV infection.

SU-FF-J-104: In Vitro Model to Study the Biological Effect of Dose Gradients On a Cell Culture System

2005 ◽  
Vol 32 (6Part6) ◽  
pp. 1944-1944 ◽  
Author(s):  
R Bromley ◽  
L Oliver ◽  
R Harvie ◽  
R Davey
Keyword(s):  
Cell Culture ◽  
Biological Effect ◽  
Culture System ◽  
In Vitro Model ◽  
Cell Culture System ◽  
A Cell ◽  
Vitro Model

Production of monoclonal antibodies against hepatitis E virus capsid protein and evaluation of their neutralizing activity in a cell culture system

2008 ◽  
Vol 153 (4) ◽  
pp. 657-666 ◽  
Author(s):  
Masaharu Takahashi ◽  
Yu Hoshino ◽  
Toshinori Tanaka ◽  
Hideyuki Takahashi ◽  
Tsutomu Nishizawa ◽  
...  
Keyword(s):  
Cell Culture ◽  
Monoclonal Antibodies ◽  
Capsid Protein ◽  
Culture System ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Cell Culture System ◽  
Virus Capsid ◽  
A Cell ◽  
Virus Capsid Protein

Generation of a Bactrian camel hepatitis E virus by a reverse genetics system

2021 ◽  
Vol 102 (7) ◽  
Author(s):  
Wenjing Zhang ◽  
Yasushi Ami ◽  
Yuriko Suzaki ◽  
Yen Hai Doan ◽  
Naokazu Takeda ◽  
...  
Keyword(s):  
Cell Culture ◽  
Culture System ◽  
Hepatitis E Virus ◽  
Reverse Genetics ◽  
Hepatitis E ◽  
Bactrian Camel ◽  
Cell Culture System ◽  
Zoonotic Infection ◽  
Cynomolgus Monkeys ◽  
A Cell

Bactrian camel hepatitis E virus (HEV) is a novel HEV belonging to genotype 8 (HEV-8) in the Orthohepevirus A species of the genus Hepevirus in the family Hepeviridae. HEV-8 cross-transmits to cynomolgus monkeys and has a potential risk for zoonotic infection. Until now, neither a cell-culture system to grow the virus nor a reverse genetics system to generate the virus has been developed. To generate replication-competent HEV-8 and to establish a cell-culture system, we synthesized capped genomic HEV-8 RNAs by in vitro transcription and used them to transfect into PLC/PRF/5 cells. A HEV-8 strain, HEV-8M2, was recovered from the capped HEV-8 RNA–transfected cell-culture supernatants and subsequently passaged in the cells, demonstrating that PLC/PRF/5 cells were capable of supporting the replication of the HEV-8, and that a cell-culture system for HEV-8 was successfully established. In addition to PLC/PRF/5 cells, A549 and Caco-2 cells appeared to be competent for the replication, but HepG2 C3/A, Vero, Hela S3, HEp-2C, 293T and GL37 cells were incompetent. The HEV-8M2 strain was capable of infecting cynomolgus monkeys by an intravenous inoculation, indicating that HEV-8 was infectious and again carried a risk for zoonotic infection. In contrast, HEV-8 did not infect nude rats and BALB/c nude mice, suggesting that the reservoir of HEV-8 was limited. In addition, the replication of the HEV-8M2 strain was efficiently abrogated by ribavirin but not by favipiravir, suggesting that ribavirin is a drug candidate for therapeutic treatment of HEV-8-induced hepatitis. The infectious HEV-8 produced by a reverse genetics system would be useful to elucidate the mechanisms of HEV replication and the pathogenesis of type E hepatitis.

scholarly journals Isospora suis in an Epithelial Cell Culture System – An In Vitro Model for Sexual Development in Coccidia

PLoS ONE ◽  
2013 ◽  
Vol 8 (7) ◽  
pp. e69797 ◽  
Author(s):  
Hanna Lucia Worliczek ◽  
Bärbel Ruttkowski ◽  
Lukas Schwarz ◽  
Kirsti Witter ◽  
Waltraud Tschulenk ◽  
...  
Keyword(s):  
Cell Culture ◽  
Epithelial Cell ◽  
Sexual Development ◽  
Culture System ◽  
In Vitro Model ◽  
Cell Culture System ◽  
Epithelial Cell Culture ◽  
Isospora Suis ◽  
Vitro Model

O.029 Analysis of hepatitis E virus mutants in a new cell culture system

2006 ◽  
Vol 36 ◽  
pp. S9
Author(s):  
S.U. Emerson ◽  
H. Nguyen ◽  
U. Torian ◽  
R.H. Purcell
Keyword(s):  
Cell Culture ◽  
Culture System ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Cell Culture System

scholarly journals Development and evaluation of an efficient cell-culture system for Hepatitis E virus

2007 ◽  
Vol 88 (3) ◽  
pp. 903-911 ◽  
Author(s):  
Toshinori Tanaka ◽  
Masaharu Takahashi ◽  
Eiji Kusano ◽  
Hiroaki Okamoto
Keyword(s):  
Cell Culture ◽  
Culture System ◽  
Hepatitis E Virus ◽  
Culture Medium ◽  
Hepatitis E ◽  
Post Inoculation ◽  
Cell Culture System ◽  
Genotype 3 ◽  
Serum Samples ◽  
Efficient Cell Culture System

Using a faecal suspension with high load of Hepatitis E virus (HEV) (2.0×107 copies ml−1, genotype 3), we developed an efficient cell-culture system for HEV in a hepatocarcinoma cell line (PLC/PRF/5). HEV progeny released in the culture medium were passaged five times successively in PLC/PRF/5 cells. The initial day of appearance and load of HEV detectable in the culture supernatant after inoculation were dependent on the titre of seed virus in the inoculum. When 6.4×104 copies of HEV were inoculated on monolayers of PLC/PRF/5 cells in six-well microplates, HEV RNA was first detected in the culture medium on day 14 post-inoculation and increased to 9.1×105 copies ml−1 on day 60. When 8.6×105 copies of HEV were inoculated, HEV RNA was initially detected on day 12 and reached the highest titre of 8.6×107 copies ml−1 on day 60. HEV incubated at temperatures higher than 70 °C did not grow in PLC/PRF/5 cells, while HEV incubated at 56 °C for 30 min was infectious. Convalescent serum samples with IgM-class HEV antibodies obtained from patients infected with HEV of genotype 1, 3 or 4 neutralized the genotype 3 virus, indicating that HEV antibodies are broadly cross-reactive. Serum samples obtained from patients 8.7 or 24.0 years after the onset of HEV infection also prevented the propagation of HEV in PLC/PRF/5 cells, suggesting the presence of long-lasting HEV antibodies with neutralizing activity in individuals with past HEV infection.

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