title: [Establishment of a cell-based 2009 H1N1 influenza neuraminidase inhibitors evaluation system].
abstract: This study is to establish a cell-based model targeting to neuraminidase (NA) of the 2009 H1N1 influenza A virus. NA is an influenza virus structural protein with enzymatic activity of the cleavage of HA-sialic acid interaction to release new viral particles from cells. A model of HIV-1 (pNL4-3.Luc.R(-)E(-)) based pseudovirions packed with HA [hemagglutinin, A/VietNam/1203/2004 (H5N1)] and NA [A/California/04/2009 (H1N1)] was established to evaluate compounds activities on NA function. The viral release can be blocked by neuraminidase inhibitors, oseltamivir and oseltamivir carboxylate, with IC50 of (61 +/- 31) nmol L(-1) and (5.5 +/- 2.9) nmol L(-1) respectively. A point mutation of H275Y on NA leads oseltamivir-resistance. This corresponding mutation was introduced into the system which was also confirmed by oseltamivir and oseltamivir carboxylate.
authors.affiliation: Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Zhang
authors.forename: Ying-li
authors.initials: YL
authors.lastname: Cao
authors.forename: Wu
authors.initials: W
authors.lastname: Zhong
authors.forename: Jun-hai
authors.initials: JH
authors.lastname: Xiao
authors.forename: Ying
authors.initials: Y
authors.lastname: Guo
mesh_terms: D045744:Cell Line, Tumor; D024882:Drug Resistance, Viral / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D057809:HEK293 Cells; D015497:HIV-1 / Q000235:genetics; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics / Q000378:metabolism; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D053124:Influenza A Virus, H5N1 Subtype / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D009439:Neuraminidase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D053139:Oseltamivir / Q000031:analogs & derivatives* / Q000494:pharmacology*; D010957:Plasmids; D014162:Transfection; D053586:Virus Internalization
chemical_list: D004791:Enzyme Inhibitors; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D053139:Oseltamivir; D009439:Neuraminidase; C535162:oseltamivir carboxylate