title: In vitro inhibition of hepatitis C virus by antisense oligonucleotides in PBMC compared to hepatoma cells.
abstract: OBJECTIVE
  To assess the efficiency of phosphorothioate antisense oligodeoxynucleotide 1 (S-ODN1) on HCV translation inhibition in PBMC compared to hepatoma cells in vitro for the first time.
  
  
  METHODS
  The study included 34 treatment naive HCV patients. IRES domain III and IV sequence variations were tested in 45 clones from 9 HCV patients. PBMC of HCV positive patients were subjected to S-ODN in vitro. Concomitantly HepG2 cells infected by the same patient's serum were also treated with S-ODN1 for 24 and 48 hours. Cellular RNA was tested for HCV plus and minus strands by reverse transcription polymerase chain reaction (RT-PCR).
  
  
  RESULTS
  Sequence variations were seen in HCV IRES domain III only while domain IV was conserved among all the tested patient's clones. S-ODN1 successfully inhibited HCV translation in HepG2 cells, while in PBMC inhibition was partial.
  
  
  CONCLUSIONS
  HCV IRES domain IV is more conserved than domain IIId in genotype 4 HCV patients. S-ODN against HCV IRES domain IV was not efficient to inhibit HCV translation in PBMC under the study conditions. Further studies testing other S-ODN targeting other HCV IRES domains in PBMC should be done.
authors.affiliation: Microbial Biotechnology Department, National Research Center, Cairo 12311, Egypt.
authors.forename: Samar Samir
authors.initials: SS
authors.lastname: Youssef
authors.affiliation: Reproductive Health and Family Planning Department, National Research Center, Cairo, Egypt ; INRS-Institut Armand Frappier, Laval, QC, Canada H7V 1B7.
authors.forename: Ahmed Mohamed
authors.initials: AM
authors.lastname: Fahmy
authors.affiliation: Microbial Biotechnology Department, National Research Center, Cairo 12311, Egypt.
authors.forename: Moataza Hassan
authors.initials: MH
authors.lastname: Omran
authors.affiliation: Chemistry Department, Faculty of Science, Cairo University, Cairo 12613, Egypt.
authors.forename: Amr Saad
authors.initials: AS
authors.lastname: Mohamed
authors.affiliation: Chemistry Department, Faculty of Science, Cairo University, Cairo 12613, Egypt.
authors.forename: Mohamed Ali
authors.initials: MA
authors.lastname: El Desouki
authors.affiliation: Microbial Biotechnology Department, National Research Center, Cairo 12311, Egypt.
authors.forename: Mostafa K
authors.initials: MK
authors.lastname: El-Awady
mesh_terms: D020121:5' Untranslated Regions / Q000235:genetics; D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000473:pathology / Q000628:therapy / Q000821:virology; D056945:Hep G2 Cells; D016174:Hepacivirus / Q000187:drug effects / Q000235:genetics*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology; D008113:Liver Neoplasms / Q000235:genetics* / Q000473:pathology / Q000628:therapy / Q000821:virology; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000235:genetics*; D014176:Protein Biosynthesis; D012367:RNA, Viral / Q000037:antagonists & inhibitors / Q000235:genetics; D053586:Virus Internalization
chemical_list: D020121:5' Untranslated Regions; D016376:Oligonucleotides, Antisense; D012367:RNA, Viral