title: New evidence of replication of hepatitis C virus in short-term peripheral blood mononuclear cell cultures.
abstract: BACKGROUND
  Even though hepatocytes are the main site for hepatitis C virus (HCV) replication, peripheral blood mononuclear cells (PBMC) have also been proposed as a suitable site for HCV replication. However, this issue still remains under discussion. We have previously developed an innovative system where HCV-RNA can be recovered during PBMC culture from HCV infected patients. Thus, the aim of this work was to use this novel approach in order to observe the evolution and replication of HCV genotype 1b in the PBMC of an HIV-HCV coinfected patient.
  
  
  METHODS
  HCV-RNA was extracted from serum, uncultured PBMC and PBMC culture at day 6, 20 and 33. The evolutionary analysis was performed using the direct sequences of three viral regions: 5'UTR, E2 and NS5A. Additionally, E2 region was cloned in order to extend the evolutive analysis.
  
  
  RESULTS
  In the present work, the molecular characterization of HCV along the culture showed a clear dynamic evolving process with the appearance of several nucleotide or amino acid changes in the three regions analyzed. Furthermore, the population analysis of E2 clones showed emerging and loss of lineages which indicate the fast evolutive dynamics of this system.
  
  
  CONCLUSIONS
  Since evolution can take place only if the virus is replicating in the culture, this finding constitutes an important evidence of viral replication in PBMC. Moreover, this extrahepatic compartment could be very important due to the presence of distinctive variants that could be responsible for resistance to treatment, viral pathogenesis and other clinical implications.
authors.affiliation: Cátedra de Virología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina. Electronic address: fadilello@ffyb.uba.ar.
authors.forename: Federico Alejandro
authors.initials: FA
authors.lastname: Di Lello
authors.affiliation: Cátedra de Virología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina.
authors.forename: Andrés Carlos Alberto
authors.initials: AC
authors.lastname: Culasso
authors.affiliation: Instituto de Investigaciones Hematológicas "Mariano R. Castex", Instituto de Medicina Experimental (IMEx), Academia Nacional de Medicina, Ciudad Autónoma de Buenos Aires, Argentina.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Parodi
authors.affiliation: Instituto de Investigaciones Hematológicas "Mariano R. Castex", Instituto de Medicina Experimental (IMEx), Academia Nacional de Medicina, Ciudad Autónoma de Buenos Aires, Argentina.
authors.forename: Patricia
authors.initials: P
authors.lastname: Baré
authors.affiliation: Cátedra de Virología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina.
authors.forename: Rodolfo Héctor
authors.initials: RH
authors.lastname: Campos
authors.affiliation: Cátedra de Virología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina.
authors.forename: Gabriel
authors.initials: G
authors.lastname: García
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D002478:Cells, Cultured; D005500:Follow-Up Studies; D016174:Hepacivirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D006526:Hepatitis C / Q000821:virology*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology*; D008297:Male; D008969:Molecular Sequence Data; D010802:Phylogeny; D016415:Sequence Alignment; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics; D014776:Virus Cultivation; D014779:Virus Replication; D055815:Young Adult
chemical_list: D014764:Viral Proteins
keywords: Bayesian; Evolution; HCV; PBMC; Phylogenetic