title: Influenza virus utilizes N-linked sialoglycans as receptors in A549 cells.
abstract: Influenza viruses (IFVs) recognize sialoglycans expressed on the host cell surface. To understand the mechanisms underlying tissue and host tropisms of IFV, it is essential to elucidate the molecular interaction of the virus with the host sialoglycan receptor. We established and applied a new monoclonal antibody, clone HYB4, which specifically recognizes the Neu5Acα2-3 determinant at the non-reducing terminal Gal residue of both glycoproteins and gangliosides to investigate the biochemical properties of IFV receptors in A549 cells. HYB4 significantly blocked virus binding to A549 cells in a dose-dependent manner. Virus overlay assay indicated that several glycoproteins with molecular masses of 80-120 kDa of A549 cells were commonly recognized by different subtypes of IFV, such as H1N1 and H3N2. H1N1 virus binding to the glycoproteins was diminished by pretreatment with either sialidase or PNGase F. On TLC-immunostaining experiments with HYB4, GM₃ ganglioside was only detected in A549 cells. Interestingly, this antibody bound to GM₃ gangliosides on TLC and plastic surfaces, but not on lipid bilayers. In comparison with the recognition of Maackia amurensis lectins, HYB4 exclusively recognized Neu5Acα2-3Galβ1-4GlcNAc residues expressed on glycoproteins. These results strongly suggest that N-linked sialoglycans with the Neu5Acα2-3 determinant on several glycoproteins are receptors for influenza virus in A549 cells.
authors.affiliation: Department of Biochemistry, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka-shi, Shizuoka 422-8526, Japan. hidari@jc.u-aizu.ac.jp
authors.forename: Kazuya I P J
authors.initials: KI
authors.lastname: Hidari
authors.forename: Maho
authors.initials: M
authors.lastname: Yamaguchi
authors.forename: Fumihiko
authors.initials: F
authors.lastname: Ueno
authors.forename: Tomoko
authors.initials: T
authors.lastname: Abe
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Yoshida
authors.forename: Takashi
authors.initials: T
authors.lastname: Suzuki
mesh_terms: D000911:Antibodies, Monoclonal / Q000378:metabolism / Q000494:pharmacology; D000918:Antibody Specificity; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D005679:G(M3) Ganglioside / Q000378:metabolism; D054884:Host-Pathogen Interactions; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000378:metabolism*; D053122:Influenza A Virus, H3N2 Subtype / Q000378:metabolism*; D008051:Lipid Bilayers / Q000378:metabolism; D029901:Maackia / Q000737:chemistry; D008562:Membrane Glycoproteins / Q000378:metabolism; D008970:Molecular Weight; D009439:Neuraminidase / Q000494:pharmacology; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000494:pharmacology; D010835:Phytohemagglutinins / Q000378:metabolism; D037121:Plant Lectins / Q000378:metabolism; D011956:Receptors, Cell Surface; D011991:Receptors, Virus / Q000378:metabolism*; D054788:Ribosome Inactivating Proteins / Q000378:metabolism; D053585:Virus Attachment / Q000187:drug effects
chemical_list: D000911:Antibodies, Monoclonal; D005679:G(M3) Ganglioside; D008051:Lipid Bilayers; D008562:Membrane Glycoproteins; D010835:Phytohemagglutinins; D037121:Plant Lectins; D011956:Receptors, Cell Surface; D011991:Receptors, Virus; C050600:Sambucus nigra lectins; C100156:leukoagglutinins, plants; C092488:sialic acid receptor; D009439:Neuraminidase; D054788:Ribosome Inactivating Proteins; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
keywords: HA; IFV; Influenza virus; MAH; MAM; Maackia amurensis hemagglutinin; Maackia amurensis mitogen; Monoclonal antibody; N-acetylneuraminic acid; N-glycolylneuraminic acid; Neu5Ac; Neu5Gc; Receptor; SPR; SSA; Sambucus sieboldiana agglutinin; Sia; Siaα2-3 glycans; TLC; hemagglutinin; influenza virus; sialic acid; surface plasmon resonance; thin-layer chromatography