title: Cell fusion assay by expression of respiratory syncytial virus (RSV) fusion protein to analyze the mutation of palivizumab-resistant strains.
abstract: Respiratory syncytial virus (RSV) consists of fusion (F), glyco (G), and small hydrophobic (SH) proteins as envelope proteins, and infects through cell fusion. F protein is expressed on the surface of infected cells, and induces cell fusion. In the present report, expression plasmids of the F, G and SH proteins were constructed and cell fusion activity was investigated under T7 RNA polymerase. F protein alone induced cell fusion at a lower concentration than previously reported, and co-expression of F and SH proteins induced cell fusion more efficiently than F protein alone. Palivizumab is the only prophylactic agent against RSV infection. Palivizumab-resistant strains having mutations of the F protein of K272E and S275F were reported. These mutations were introduced into an F-expression plasmid, and exhibited no inhibition of cell fusion with palivizumab. Among the RSV F protein mutants, N276S has been reported to have partial resistance against palivizumab, but the F expression plasmid with the N276S mutation showed a reduction in cell fusion in the presence of palivizumab, showing no resistance to palivizumab. The present expression system was useful for investigating the mechanisms of RSV cell fusion.
authors.affiliation: Keio University, Health Center, 4-1-1 Hiyoshi, Kouhoku, Yokohama, Kanagawa, Japan; Laboratory of Viral Infection I, Kitasato Institute for Life Sciences, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo, Japan. Electronic address: yyasui@keio.jp.
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Yasui
authors.affiliation: Laboratory of Viral Infection I, Kitasato Institute for Life Sciences, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo, Japan. Electronic address: di12004@st.kitasato-u.ac.jp.
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Yamaji
authors.affiliation: Laboratory of Viral Infection I, Kitasato Institute for Life Sciences, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo, Japan. Electronic address: akihito@lisci.kitasato-u.ac.jp.
authors.forename: Akihito
authors.initials: A
authors.lastname: Sawada
authors.affiliation: Laboratory of Viral Infection I, Kitasato Institute for Life Sciences, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo, Japan. Electronic address: takashinemo@yahoo.co.jp.
authors.forename: Takashi
authors.initials: T
authors.lastname: Ito
authors.affiliation: Laboratory of Viral Infection I, Kitasato Institute for Life Sciences, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo, Japan. Electronic address: tetsuo-n@lisci.kitasato-u.ac.jp.
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Nakayama
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002459:Cell Fusion; D002460:Cell Line; D003584:Cytological Techniques / Q000379:methods; D024882:Drug Resistance, Viral; D006801:Humans; D009154:Mutation; D000069455:Palivizumab / Q000494:pharmacology*; D012136:Respiratory Syncytial Viruses / Q000187:drug effects* / Q000235:genetics; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism; D014760:Viral Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014773:Virology / Q000379:methods
chemical_list: D000998:Antiviral Agents; C526850:F protein, human respiratory syncytial virus; C502275:G glycoprotein, Respiratory syncytial virus; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins; D000069455:Palivizumab
keywords: Cell fusion; F protein; G protein; Palivizumab resistant strain; Respiratory syncytial virus; SH protein