HA-pseudotyped retroviral vectors for influenza antagonist screening.
Influenza infections are initiated by the binding of the influenza hemagglutinin (HA) and the cellular receptor sialic acids. The binding is followed by internalization, endocytosis, and uncoating to release the influenza genome to the cytoplasm. It is conceivable that specific inhibitors that antagonize any one of these events could prevent the replication of influenza infections. The authors made HA pseudotyped retroviral vectors that express luciferase reporter activities upon transduction to several recipient cells. The transduction of the HA-pseudotype virus particles (HApp) was mediated through the specific interactions between an avian HA and the terminal disaccharides of sialic acid (SA) and galactose (Gal) in alpha-2,3 linkage. The HApp-mediated transduction method was used to develop a high-throughput screening assay and to screen for hits from a fermentation extract library. Specific hits that inhibited the HA-mediated but were noninhibitory to the vesicular stomatitis virus-mediated pseudoviral transductions were identified. A few of these hits have anti-influenza activities that prevent the replication of both H1N1 (WSN) and H5N1 (RG14) influenza viruses.
Wang
Shi-Yun
SY
Genomics Research Center, Academia Sinica, Taipei, Taiwan, Republic of China.
Su
Ching-Yao
CY
Lin
Mengi
M
Huang
Shao-Yung
SY
Huang
Wen-I
WI
Wang
Cheng-Chi
CC
Wu
Ying-Ta
YT
Cheng
Ting-Jen R
TJ
Yu
Hui-Ming
HM
Ren
Chien-Tai
CT
Wu
Chung-Yi
CY
Wong
Chi-Huey
CH
Cheng
Yih-Shyun E
YS
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D045744:Cell Line, Tumor; D004285:Dogs; D004353:Drug Evaluation, Preclinical; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D007668:Kidney / Q000166:cytology; D008156:Luciferases / Q000378:metabolism; D008175:Lung Neoplasms / Q000473:pathology; D010957:Plasmids; D011994:Recombinant Proteins / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics; D014161:Transduction, Genetic; D014162:Transfection; D014779:Virus Replication / Q000187:drug effects
D000998:Antiviral Agents; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D011994:Recombinant Proteins; D008156:Luciferases