title: Neutralizing monoclonal antibodies to different clades of Influenza A H5N1 viruses.
abstract: Four IgG(1kappa) monoclonal antibodies (mAbs) against Influenza A/Chicken/Vietnam/8/2004 (H5N1) virus are described. Three of these showed neutralizing activities against H5N1 strains from clades 1, 2 and 3 using a retroviral pseudotype or live virus microneutralization assay. In the pseudotype assay, the IC(90) neutralizing titre range was >1600-51,200, and with the microneutralization was 80> or =10,240. MAb 1C1 showed strong neutralizing activities in both assays. All four mAbs reacted specifically to the immunogen by immunohistochemical staining and to A/Hong Kong/483/1997 (H5N1) and A/Thailand/1(KAN-1)/2004 (H5N1)-infected MDCK cells by immunofluorescence. ELISA titrations of the mAbs showed specificity for H5N1 haemagglutinin (HA) and no cross-reactivity to 15 other Influenza A subtypes. Only mAbs 1C1 and the non-neutralizing 1F7 reacted with HA(1), the cleaved subunit of HA, by Western blot. These results suggest that the mAbs recognize distinct or overlapping epitopes and will be useful reagents for construction of specific rapid point-of-care assays or for therapeutic use.
authors.affiliation: Institute of Medical and Veterinary Science, Adelaide, SA, Australia. sawyin.oh@imvs.sa.gov.au
authors.forename: Sawyin
authors.initials: S
authors.lastname: Oh
authors.forename: Paul
authors.initials: P
authors.lastname: Selleck
authors.forename: Nigel J
authors.initials: NJ
authors.lastname: Temperton
authors.forename: Paul K S
authors.initials: PK
authors.lastname: Chan
authors.forename: Barbara
authors.initials: B
authors.lastname: Capecchi
authors.forename: Jim
authors.initials: J
authors.lastname: Manavis
authors.forename: Geoff
authors.initials: G
authors.lastname: Higgins
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Burrell
authors.forename: Tuckweng
authors.initials: T
authors.lastname: Kok
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000276:immunology* / Q000302:isolation & purification; D002645:Chickens; D003429:Cross Reactions; D006389:Hemagglutinins, Viral / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology / Q000302:isolation & purification; D053124:Influenza A Virus, H5N1 Subtype / Q000276:immunology* / Q000302:isolation & purification; D020128:Inhibitory Concentration 50; D009500:Neutralization Tests
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D006389:Hemagglutinins, Viral; D007074:Immunoglobulin G; C458881:hemagglutinin fusogenic peptide, influenza virus