Cell fusion assay by expression of respiratory syncytial virus (RSV) fusion protein to analyze the mutation of palivizumab-resistant strains

J Virol Methods. 2016 May:231:48-55. doi: 10.1016/j.jviromet.2016.01.003. Epub 2016 Jan 12.

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.

Keywords: Cell fusion; F protein; G protein; Palivizumab resistant strain; Respiratory syncytial virus; SH protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antiviral Agents / pharmacology*
  • Cell Fusion*
  • Cell Line
  • Cytological Techniques / methods
  • Drug Resistance, Viral*
  • Humans
  • Mutation*
  • Palivizumab / pharmacology*
  • Respiratory Syncytial Viruses / drug effects*
  • Respiratory Syncytial Viruses / genetics
  • Viral Envelope Proteins / genetics*
  • Viral Envelope Proteins / metabolism
  • Viral Fusion Proteins / genetics
  • Viral Fusion Proteins / metabolism
  • Virology / methods

Substances

  • Antiviral Agents
  • F protein, human respiratory syncytial virus
  • G glycoprotein, Respiratory syncytial virus
  • Viral Envelope Proteins
  • Viral Fusion Proteins
  • Palivizumab