TY - JOUR
T1 - Influenza virus utilizes N-linked sialoglycans as receptors in A549 cells
AU - Hidari, Kazuya I.P.J.
AU - Yamaguchi, Maho
AU - Ueno, Fumihiko
AU - Abe, Tomoko
AU - Yoshida, Kazuhiro
AU - Suzuki, Takashi
N1 - Funding Information:
We thank Dr. Taiichi Usui, Dr. Takeomi Murata, and Dr. Makoto Ogata for their gifts of glycopolymers. This work was supported by grants-in-aid for Scientific Research on Priority Areas ( 24570168 to K.I.P.J.H.) from the Ministry of Education, Science, Sports, and Culture of Japan . This work was also supported by research grants from Heiwa Nakajima Foundation and Japan China Medical Association .
PY - 2013/7/5
Y1 - 2013/7/5
N2 - 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 80-120kDa 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.
AB - 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 80-120kDa 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.
KW - Influenza virus
KW - Monoclonal antibody
KW - Receptor
KW - Siaα2-3 glycans
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U2 - 10.1016/j.bbrc.2013.05.112
DO - 10.1016/j.bbrc.2013.05.112
M3 - Article
C2 - 23747419
AN - SCOPUS:84879882070
SN - 0006-291X
VL - 436
SP - 394
EP - 399
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 3
ER -