TY - JOUR
T1 - Generation of transmissible hepatitis C virions from a molecular clone in chimpanzees
AU - Hong, Zhi
AU - Beaudet-Miller, Michele
AU - Lanford, Robert E.
AU - Guerra, Bernadette
AU - Wright-Minogue, Jacquelyn
AU - Skelton, Angela
AU - Baroudy, Bahige M.
AU - Reyes, Gregory R.
AU - Lau, Johnson Y.N.
N1 - Funding Information:
We are grateful to Drs. Alfred Prince and Charles Rice for their support and helpful discussion and for generously providing their molecular clones. We thank Dr. Gene B. Hubbard for the evaluation of the liver biopsies and for generating the pathology reports. This work was also supported by a research grant to R.E.L. from Schering-Plough Research Institute (Kenilworth, NJ).
PY - 1999/3/30
Y1 - 1999/3/30
N2 - Multiple alignments of hepatitis C virus (HCV) polyproteins from six different genotypes identified a total of 22 nonconsensus mutations in a clone derived from the Hutchinson (H77) isolate. These mutations, collectively, may have contributed to the failure in generating a 'functionally correct' or 'infectious' clone in earlier attempts. A consensus clone was constructed after systematic repair of these mutations, which yielded infectious virions in a chimpanzee after direct intrahepatic inoculation of in vitro transcribed RNAs. This RNA-infected chimpanzee has developed hepatitis and remained HCV positive for more than 11 months. To further verify this RNA-derived infectivity, a second naive chimpanzee was injected intravenously with serum collected from the first chimpanzee. Infectivity analysis of the second chimpanzee demonstrated that the HCV infection was successfully transmitted, which validated unequivocally the infectivity of our repaired molecular clone. Amino acid sequence comparisons revealed that our repaired infectious clone had 4 mismatches with the isogenic clone reported by Kolykhalov et al. (1997, Science 277, 570-574) and 8 mismatches with that reported by Yanagi et al. (1997, Proc. Natl. Acad. Sci. USA 94, 8738-8743). At the RNA level, more mismatches (43 and 67, respectively) were identified; most of them were synonymous substitutions. Further comparisons with 16 isolates from different genotypes demonstrated that our repaired clone shares greater consensus than the reported isogenic clones. This approach of generating infectious HCV RNA validates the importance of amino acid sequence consensus in relation to the biology of HCV.
AB - Multiple alignments of hepatitis C virus (HCV) polyproteins from six different genotypes identified a total of 22 nonconsensus mutations in a clone derived from the Hutchinson (H77) isolate. These mutations, collectively, may have contributed to the failure in generating a 'functionally correct' or 'infectious' clone in earlier attempts. A consensus clone was constructed after systematic repair of these mutations, which yielded infectious virions in a chimpanzee after direct intrahepatic inoculation of in vitro transcribed RNAs. This RNA-infected chimpanzee has developed hepatitis and remained HCV positive for more than 11 months. To further verify this RNA-derived infectivity, a second naive chimpanzee was injected intravenously with serum collected from the first chimpanzee. Infectivity analysis of the second chimpanzee demonstrated that the HCV infection was successfully transmitted, which validated unequivocally the infectivity of our repaired molecular clone. Amino acid sequence comparisons revealed that our repaired infectious clone had 4 mismatches with the isogenic clone reported by Kolykhalov et al. (1997, Science 277, 570-574) and 8 mismatches with that reported by Yanagi et al. (1997, Proc. Natl. Acad. Sci. USA 94, 8738-8743). At the RNA level, more mismatches (43 and 67, respectively) were identified; most of them were synonymous substitutions. Further comparisons with 16 isolates from different genotypes demonstrated that our repaired clone shares greater consensus than the reported isogenic clones. This approach of generating infectious HCV RNA validates the importance of amino acid sequence consensus in relation to the biology of HCV.
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U2 - 10.1006/viro.1999.9603
DO - 10.1006/viro.1999.9603
M3 - Article
C2 - 10087224
AN - SCOPUS:0033616519
VL - 256
SP - 36
EP - 44
JO - Virology
JF - Virology
SN - 0042-6822
IS - 1
ER -