PMID- 23151512
OWN - NLM
STAT- MEDLINE
DCOM- 20130219
LR  - 20211021
IS  - 1091-6490 (Electronic)
IS  - 0027-8424 (Print)
IS  - 0027-8424 (Linking)
VI  - 109
IP  - 48
DP  - 2012 Nov 27
TI  - Highly efficient full-length hepatitis C virus genotype 1 (strain TN) infectious 
      culture system.
PG  - 19757-62
LID - 10.1073/pnas.1218260109 [doi]
AB  - Chronic infection with hepatitis C virus (HCV) is an important cause of end stage 
      liver disease worldwide. In the United States, most HCV-related disease is 
      associated with genotype 1 infection, which remains difficult to treat. Drug and 
      vaccine development was hampered by inability to culture patient isolates 
      representing HCV genotypes 1-7 and subtypes; only a recombinant 2a genome (strain 
      JFH1) spontaneously replicated in vitro. Recently, we identified three mutations 
      F1464L/A1672S/D2979G (LSG) in the nonstructural (NS) proteins, essential for 
      development of full-length HCV 2a (J6) and 2b (J8) culture systems in Huh7.5 
      cells. Here, we developed a highly efficient genotype 1a (strain TN) full-length 
      culture system. We initially found that the LSG substitutions conferred viability 
      to an intergenotypic recombinant composed of TN 5' untranslated region 
      (5'UTR)-NS5A and JFH1 NS5B-3'UTR; recovered viruses acquired two adaptive 
      mutations located in NS3 and NS4B. Introduction of these changes into a 
      replication-deficient TN full-length genome, harboring LSG, permitted efficient 
      HCV production. Additional identified NS4B and NS5B mutations fully adapted the 
      TN full-length virus. Thus, a TN genome with 8 changes (designated TN 
      cell-culture derived, TNcc) replicated efficiently and released infectious 
      particles of  approximately 5 log(10) focus-forming units per mL; passaged TNcc did not require 
      additional changes. IFN-alpha and directly acting antivirals targeting the HCV 
      protease, NS5A, and NS5B, each inhibited full-length TN infection 
      dose-dependently. Given the unique importance of genotype 1 for pathogenesis, 
      this infectious 1a culture system represents an important advance in HCV 
      research. The approach used and the mutations identified might permit culture 
      development for other HCV isolates, thus facilitating vaccine development and 
      personalized treatment.
FAU - Li, Yi-Ping
AU  - Li YP
AD  - Copenhagen Hepatitis C Program, Department of Infectious Diseases and Clinical 
      Research Centre, Copenhagen University Hospital, DK-2650 Hvidovre, Denmark.
FAU - Ramirez, Santseharay
AU  - Ramirez S
FAU - Jensen, Sanne B
AU  - Jensen SB
FAU - Purcell, Robert H
AU  - Purcell RH
FAU - Gottwein, Judith M
AU  - Gottwein JM
FAU - Bukh, Jens
AU  - Bukh J
LA  - eng
SI  - GENBANK/JX993348
GR  - Intramural NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Intramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20121114
PL  - United States
TA  - Proc Natl Acad Sci U S A
JT  - Proceedings of the National Academy of Sciences of the United States of America
JID - 7505876
RN  - 0 (3' Untranslated Regions)
RN  - 0 (NS3 protein, flavivirus)
RN  - 0 (NS4B protein, flavivirus)
RN  - 0 (Viral Nonstructural Proteins)
RN  - EC 3.4.21.- (Serine Endopeptidases)
RN  - EC 3.6.4.13 (RNA Helicases)
SB  - IM
MH  - 3' Untranslated Regions
MH  - Cell Line, Tumor
MH  - Genotype
MH  - Hepacivirus/*genetics/pathogenicity/physiology
MH  - Humans
MH  - Molecular Sequence Data
MH  - Mutation
MH  - RNA Helicases/genetics
MH  - Recombination, Genetic
MH  - Serine Endopeptidases/genetics
MH  - Viral Nonstructural Proteins/genetics
MH  - Virus Replication
PMC - PMC3511766
COIS- The authors declare no conflict of interest.
EDAT- 2012/11/16 06:00
MHDA- 2013/02/21 06:00
PMCR- 2012/11/14
CRDT- 2012/11/16 06:00
PHST- 2012/11/16 06:00 [entrez]
PHST- 2012/11/16 06:00 [pubmed]
PHST- 2013/02/21 06:00 [medline]
PHST- 2012/11/14 00:00 [pmc-release]
AID - 1218260109 [pii]
AID - 201218260 [pii]
AID - 10.1073/pnas.1218260109 [doi]
PST - ppublish
SO  - Proc Natl Acad Sci U S A. 2012 Nov 27;109(48):19757-62. doi: 
      10.1073/pnas.1218260109. Epub 2012 Nov 14.