title: Preparation of quadri-subtype influenza virus-like particles using bovine immunodeficiency virus gag protein.
abstract: Influenza VLPs comprised of hemagglutinin (HA), neuraminidase (NA), and matrix (M1) proteins have been previously used for immunological and virological studies. Here we demonstrated that influenza VLPs can be made in Sf9 cells by using the bovine immunodeficiency virus gag (Bgag) protein in place of M1. We showed that Bgag can be used to prepare VLPs for several influenza subtypes including H1N1 and H10N8. Furthermore, by using Bgag, we prepared quadri-subtype VLPs, which co-expressed within the VLP the four HA subtypes derived from avian-origin H5N1, H7N9, H9N2 and H10N8 viruses. VLPs showed hemagglutination and neuraminidase activities and reacted with specific antisera. The content and co-localization of each HA subtype within the quadri-subtype VLP were evaluated. Electron microscopy showed that Bgag-based VLPs resembled influenza virions with the diameter of 150-200nm. This is the first report of quadri-subtype design for influenza VLP and the use of Bgag for influenza VLP preparation.
authors.affiliation: Medigen, Inc., 8420 Gas House Pike, Suite S, Frederick, MD, USA.
authors.forename: Irina
authors.initials: I
authors.lastname: Tretyakova
authors.affiliation: Medigen, Inc., 8420 Gas House Pike, Suite S, Frederick, MD, USA.
authors.forename: Rachmat
authors.initials: R
authors.lastname: Hidajat
authors.affiliation: Medigen, Inc., 8420 Gas House Pike, Suite S, Frederick, MD, USA.
authors.forename: Garrett
authors.initials: G
authors.lastname: Hamilton
authors.affiliation: Medigen, Inc., 8420 Gas House Pike, Suite S, Frederick, MD, USA.
authors.forename: Noah
authors.initials: N
authors.lastname: Horn
authors.affiliation: Medigen, Inc., 8420 Gas House Pike, Suite S, Frederick, MD, USA.
authors.forename: Brian
authors.initials: B
authors.lastname: Nickols
authors.affiliation: Medigen, Inc., 8420 Gas House Pike, Suite S, Frederick, MD, USA.
authors.forename: Raphael O
authors.initials: RO
authors.lastname: Prather
authors.affiliation: Influenza Division, Centers for Disease Control and Prevention, 1600 Clifton Road N.E., Atlanta, GA, USA.
authors.forename: Terrence M
authors.initials: TM
authors.lastname: Tumpey
authors.affiliation: Medigen, Inc., 8420 Gas House Pike, Suite S, Frederick, MD, USA. Electronic address: ppushko@medigen-usa.com.
authors.forename: Peter
authors.initials: P
authors.lastname: Pushko
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D002460:Cell Line; D015683:Gene Products, gag / Q000235:genetics / Q000276:immunology*; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology*; D016087:Immunodeficiency Virus, Bovine / Q000235:genetics / Q000276:immunology*; D065926:Influenza A Virus, H10N8 Subtype / Q000235:genetics / Q000276:immunology; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000276:immunology; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics / Q000276:immunology; D064766:Influenza A Virus, H7N9 Subtype / Q000235:genetics / Q000276:immunology; D053127:Influenza A Virus, H9N2 Subtype / Q000235:genetics / Q000276:immunology; D007313:Insecta; D009439:Neuraminidase / Q000276:immunology*; D009976:Orthomyxoviridae Infections / Q000276:immunology / Q000517:prevention & control / Q000821:virology; D061987:Sf9 Cells; D018411:Spodoptera; D058425:Vaccines, Virus-Like Particle / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D015683:Gene Products, gag; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D058425:Vaccines, Virus-Like Particle; D009439:Neuraminidase
keywords: Avian influenza; BIV gag; H10N8; H5N1; H7N9; H9N2; VLP vaccine; Virus-like particle