Cell culture replication of a genotype 1b hepatitis C virus isolate cloned from a patient who underwent liver transplantation.

The introduction of the genotype 2a isolate JFH1 was a major breakthrough in the field of hepatitis C virus (HCV), allowing researchers to study the complete life cycle of the virus in cell culture. However, fully competent culture systems encompassing the most therapeutically relevant HCV genotypes...

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Autores principales:	George Koutsoudakis, Sofia Perez-del-Pulgar, Mairene Coto-Llerena, Patricia Gonzalez, Jakub Dragun, Laura Mensa, Gonzalo Crespo, Miguel Navasa, Xavier Forns
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/7eb4ccc3cb7b41198827c0891ba989d2
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spelling	oai:doaj.org-article:7eb4ccc3cb7b41198827c0891ba989d22021-11-18T06:47:23ZCell culture replication of a genotype 1b hepatitis C virus isolate cloned from a patient who underwent liver transplantation.1932-620310.1371/journal.pone.0023587https://doaj.org/article/7eb4ccc3cb7b41198827c0891ba989d22011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21887279/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203The introduction of the genotype 2a isolate JFH1 was a major breakthrough in the field of hepatitis C virus (HCV), allowing researchers to study the complete life cycle of the virus in cell culture. However, fully competent culture systems encompassing the most therapeutically relevant HCV genotypes are still lacking, especially for the highly drug-resistant genotype 1b. For most isolated HCV clones, efficient replication in cultured hepatoma cells requires the introduction of replication-enhancing mutations. However, such mutations may interfere with viral assembly, as occurs in the case of the genotype 1b isolate Con1. In this study, we show that a clinical serum carrying a genotype 1b virus with an exceptionally high viral load was able to infect Huh7.5 cells. Similar to previous reports, inoculation of Huh7.5 cells by natural virus is very inefficient compared to infection by cell culture HCV. A consensus sequence of a new genotype 1b HCV isolate was cloned from the clinical serum (designated Barcelona HCV1), and then subjected to replication studies. This virus replicated poorly in a transient fashion in Huh7.5 cells after electroporation with in vitro transcribed RNA. Nonetheless, approximately 3 weeks post electroporation and thereafter, core protein-positive cells were detected by immunofluorescence. Surprisingly, small amounts of core protein were also measurable in the supernatant of electroporated cells, suggesting that HCV particles might be assembled and released. Our findings not only enhance the current method of cloning in vitro HCV replication-competent isolates, but also offer valuable insights for the realization of fully competent culture systems for HCV.George KoutsoudakisSofia Perez-del-PulgarMairene Coto-LlerenaPatricia GonzalezJakub DragunLaura MensaGonzalo CrespoMiguel NavasaXavier FornsPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 8, p e23587 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q George Koutsoudakis Sofia Perez-del-Pulgar Mairene Coto-Llerena Patricia Gonzalez Jakub Dragun Laura Mensa Gonzalo Crespo Miguel Navasa Xavier Forns Cell culture replication of a genotype 1b hepatitis C virus isolate cloned from a patient who underwent liver transplantation.
description	The introduction of the genotype 2a isolate JFH1 was a major breakthrough in the field of hepatitis C virus (HCV), allowing researchers to study the complete life cycle of the virus in cell culture. However, fully competent culture systems encompassing the most therapeutically relevant HCV genotypes are still lacking, especially for the highly drug-resistant genotype 1b. For most isolated HCV clones, efficient replication in cultured hepatoma cells requires the introduction of replication-enhancing mutations. However, such mutations may interfere with viral assembly, as occurs in the case of the genotype 1b isolate Con1. In this study, we show that a clinical serum carrying a genotype 1b virus with an exceptionally high viral load was able to infect Huh7.5 cells. Similar to previous reports, inoculation of Huh7.5 cells by natural virus is very inefficient compared to infection by cell culture HCV. A consensus sequence of a new genotype 1b HCV isolate was cloned from the clinical serum (designated Barcelona HCV1), and then subjected to replication studies. This virus replicated poorly in a transient fashion in Huh7.5 cells after electroporation with in vitro transcribed RNA. Nonetheless, approximately 3 weeks post electroporation and thereafter, core protein-positive cells were detected by immunofluorescence. Surprisingly, small amounts of core protein were also measurable in the supernatant of electroporated cells, suggesting that HCV particles might be assembled and released. Our findings not only enhance the current method of cloning in vitro HCV replication-competent isolates, but also offer valuable insights for the realization of fully competent culture systems for HCV.
format	article
author	George Koutsoudakis Sofia Perez-del-Pulgar Mairene Coto-Llerena Patricia Gonzalez Jakub Dragun Laura Mensa Gonzalo Crespo Miguel Navasa Xavier Forns
author_facet	George Koutsoudakis Sofia Perez-del-Pulgar Mairene Coto-Llerena Patricia Gonzalez Jakub Dragun Laura Mensa Gonzalo Crespo Miguel Navasa Xavier Forns
author_sort	George Koutsoudakis
title	Cell culture replication of a genotype 1b hepatitis C virus isolate cloned from a patient who underwent liver transplantation.
title_short	Cell culture replication of a genotype 1b hepatitis C virus isolate cloned from a patient who underwent liver transplantation.
title_full	Cell culture replication of a genotype 1b hepatitis C virus isolate cloned from a patient who underwent liver transplantation.
title_fullStr	Cell culture replication of a genotype 1b hepatitis C virus isolate cloned from a patient who underwent liver transplantation.
title_full_unstemmed	Cell culture replication of a genotype 1b hepatitis C virus isolate cloned from a patient who underwent liver transplantation.
title_sort	cell culture replication of a genotype 1b hepatitis c virus isolate cloned from a patient who underwent liver transplantation.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/7eb4ccc3cb7b41198827c0891ba989d2
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Cell culture replication of a genotype 1b hepatitis C virus isolate cloned from a patient who underwent liver transplantation.

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