Filovirus GP VSV-pseudotyped Assay Details

Pseudotyped virus was pre-incubated with KZ52 (EBOV-VSV), 16F6 (SUDV-VSV) or anti-MARV IgG (MARV)VSV for 1 hour before adding virus to vero cells overnight. After 24 hours, cells are lysed and percent of infection was determined by luciferase intensity. Neutralization was determined relative to the untreated virus controls. In dose response experiments the concentration of antibody causing 50% neutralization (IC50) will be determined.

The vesicular stomatitis virus (rVSV) whose glycoprotein gene (G) has been deleted will be used as the base platform for the pseudotype-based neutralization assay. The VSV-G glycoprotein is transiently expressed by transfection to produce virus particles. To create pseudotyped viruses the VSV-G is substituted with Ebola, Sudan or Marburgviurs GP. The system has been developed and the resulting virus (rVSV-EBOV GPΔG) can be handled at BSL-2. rVSV-EBOV GPΔG is cytopathic and expresses the firefly luciferase. Infection efficiency can be measured by quantification of luciferase.