Production of Ebola virus-like particles in Drosophila melanogaster Schneider 2 cells

J Virol Methods. 2018 Nov:261:156-159. doi: 10.1016/j.jviromet.2018.08.016. Epub 2018 Aug 23.

Abstract

In this study, we generated recombinant virus-like particles (VLPs) against family Filoviridae, genus Ebolavirus, species Zaire ebolavirus, strain Makona (EBOV) in Drosophila melanogaster Schneider 2 (S2) cells using the EBOV Makona. S2 cells were cotransfected with four viral plasmids encoding EBOV Makona proteins and protein expression was analyzed by immunoblotting. We confirmed that EBOV Makona proteins were successfully expressed in S2 cells. Additionally, we further examined the formation of intracellular and extracellular VLPs by electron microscopy. eVLPs were produced by sucrose gradient ultracentrifugation of S2 cells transfected with EBOV Makona genes, and production of VLPs was confirmed by immunoblot analysis. Collectively, our findings showed that the S2 cell system could be a promising tool for efficient production of eVLPs.

Keywords: Drosophila S2 cells; Makona; Virus-like particles; Zaire ebolavirus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Centrifugation, Density Gradient
  • Drosophila melanogaster
  • Ebolavirus / genetics*
  • Ebolavirus / ultrastructure
  • Gene Expression
  • Immunoblotting
  • Microscopy, Electron
  • Recombination, Genetic*
  • Transfection
  • Viral Proteins / analysis
  • Virosomes / genetics*
  • Virosomes / isolation & purification
  • Virosomes / metabolism*
  • Virosomes / ultrastructure

Substances

  • Viral Proteins
  • Virosomes