title: Characterization of a trypsin-dependent avian influenza H5N1-pseudotyped HIV vector system for high throughput screening of inhibitory molecules.
abstract: In this study, we have generated and characterized an avian influenza H5N1 hemagglutinin (HA), neuraminidase (NA) and M2 ion channel pseudotyped HIV-based vector system (HaNaM-pseudotyped HIV vector). The cleavage site of the HA protein was modified to necessitate trypsin-dependent maturation of the glycoprotein. HA, NA and M2 were efficiently incorporated in HIV vector particles which could transduce different cell lines in a trypsin-dependent manner. Results also showed that the presence of avian influenza M2 and NA proteins maximized both vector production and transduction and that transduction was highly sensitive to the specific NA inhibitor oseltamivir (Tamiflu). H5N1 HaNaM-pseudotyped HIV vector system was also adapted for cell-based high throughput screening of drug candidates against influenza virus infection, and its high sensitivity to the specific oseltamivir validates its potential utility in the identification of new influenza inhibitors. Overall, the trypsin-dependent H5N1-pseudotyped HIV vector can mimic avian influenza virus infection processes with sufficient precision to allow for the identification of new antivirals and to study avian influenza virus biology in a lower biosafety level laboratory environment.
authors: Laboratory of Molecular Human Retrovirology, Faculty of Medicine, University of Manitoba, Canada.
authors: Zhujun
authors: Z
authors: Ao
authors: Ami
authors: A
authors: Patel
authors: Kaylie
authors: K
authors: Tran
authors: Xinying
authors: X
authors: He
authors: Keith
authors: K
authors: Fowke
authors: Kevin
authors: K
authors: Coombs
authors: Darwyn
authors: D
authors: Kobasa
authors: Gary
authors: G
authors: Kobinger
authors: Xiaojian
authors: X
authors: Yao
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D006678:HIV / Q000235:genetics*; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D053124:Influenza A Virus, H5N1 Subtype / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D009439:Neuraminidase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D053139:Oseltamivir / Q000494:pharmacology; D014161:Transduction, Genetic; D014357:Trypsin / Q000378:metabolism*; D014764:Viral Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism
chemical_list: D000998:Antiviral Agents; D004791:Enzyme Inhibitors; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D014764:Viral Proteins; C109963:hemagglutinin, avian influenza A virus; D053139:Oseltamivir; D009439:Neuraminidase; D014357:Trypsin