Identification, molecular cloning, and analysis of full-length hepatitis C virus transmitted/founder genotypes 1, 3, and 4
Autor(es): Stoddard Mark B, Li Hui, Wang Shuyi, Saeed Mohsan, Andrus Linda, Ding Wenge, Jiang Xinpei, Learn Gerald H, von Schaewen Markus, Wen Jessica, Goepfert Paul A, Hahn Beatrice H, Ploss Alexander, Rice Charles M, Shaw George M
Resumo: Hepatitis C virus (HCV) infection is characterized by persistent replication of a complex mixture of viruses termed a quasispecies." Transmission is generally associated with a stringent population bottleneck characterized by infection by limited numbers of "transmitted/founder" (T/F) viruses. Characterization of T/F genomes of human immunodeficiency virus type 1 (HIV-1) has been integral to studies of transmission, immunopathogenesis, - vaccine development. Here, we describe the identification of complete T/F genomes of HCV by single-genome sequencing of plasma viral RNA from acutely infected subjects. A total of 2,739 single-genome-derived amplicons comprising 10,966,507 bp from 18 acute-phase - 11 chronically infected subjects were analyzed. Acute-phase sequences diversified essentially r-omly, except for the poly(U/UC) tract, which was subject to polymerase slippage. Fourteen acute-phase subjects were productively infected by more than one genetically distinct virus, permitting assessment of recombination between replicating genomes. No evidence of recombination was found among 1,589 sequences analyzed. Envelope sequences of T/F genomes lacked transmission signatures that could distinguish them from chronic infection viruses. Among chronically infected subjects, higher nucleotide substitution rates were observed in the poly(U/UC) tract than in envelope hypervariable region 1. Fourteen full-length molecular clones with variable poly(U/UC) sequences corresponding to seven genotype 1a, 1b, 3a, - 4a T/F viruses were generated. Like most unadapted HCV clones, T/F genomes did not replicate efficiently in Huh 7.5 cells, indicating that additional cellular factors or viral adaptations are necessary for in vitro replication. Full-length T/F HCV genomes - their progeny provide unique insights into virus transmission, virus evolution, - virus-host interactions associated with immunopathogenesis. Hepatitis C virus (HCV) infects 2% to 3% of the world's population - exhibits extraordinary genetic diversity. This diversity is mirrored by HIV-1, where characterization of transmitted/founder (T/F) genomes has been instrumental in studies of virus transmission, immunopathogenesis, - vaccine development. Here, we show that despite major differences in genome organization, replication strategy, - natural history, HCV (like HIV-1) diversifies essentially r-omly early in infection, - as a consequence, sequences of actual T/F viruses can be identified. This allowed us to capture by molecular cloning the full-length HCV genomes that are responsible for infecting the first hepatocytes - eliciting the initial immune responses, weeks before these events could be directly analyzed in human subjects. These findings represent an enabling experimental strategy, not only for HCV - HIV-1 research, but also for other RNA viruses of medical importance, including West Nile, chikungunya, dengue, Venezuelan encephalitis, - Ebola viruses."
Imprenta: mBio, v. 6, n. 2, p. e02518, 2015
Identificador do objeto digital: 10.1128/mBio.02518-14
Descritores: Chikungunya virus - Cell ; Chikungunya virus - DNA ; Chikungunya virus - Flaviviridae ; Chikungunya virus - Genome ; Chikungunya virus - Pathogenesis ; Chikungunya virus - RNA ; Chikungunya virus - Viral infections ; Chikungunya Virus - Virus ; Chikungunya virus - Transmission ; Chikungunya virus - Vaccine ; Chikungunya virus - Dengue
Data de publicação: 2015
