qHTS for Inhibitors of binding or entry into cells for Lassa Virus
Lassa virus is an arenavirus that is endemic in West Africa and can cause severe hemorrhagic fever in humans. Lassa virus is composed of a nucleocapsid core surrounded by a lipid bilayer containing glycoproteins that mediate host cell binding. To identify inhibitors of Lassa virus entry into cells, a pseudotype virus system was used where the nonpathogenic vesicular stomatitis virus (VSV) more ..
BioActive Compounds: 3648
Lassa virus is an arenavirus that is endemic in West Africa and can cause severe hemorrhagic fever in humans. Lassa virus is composed of a nucleocapsid core surrounded by a lipid bilayer containing glycoproteins that mediate host cell binding. To identify inhibitors of Lassa virus entry into cells, a pseudotype virus system was used where the nonpathogenic vesicular stomatitis virus (VSV) contains the Lassa virus envelop glycoproteins. This pseudotype virus, termed VSV-LV, has a Photinus luciferase reporter gene inserted in its genome. For this assay, human embryonic kidney 293 cells are incubated with VSV-LV and virus entry is detected by luminescence produced from the luciferase reporter. Small molecule inhibitors that block VSV-LV binding or entry into cells are identified by a decrease of luciferase activity.
In a collaboration between University of Texas Medical Branch in Galveston, TX and the NIH Chemical Genomics Center (NCGC) a miniaturized, high throughput, cell-based assay was developed and screened against the Molecular Libraries Small Molecule Repository (MLSMR) library.
NIH Chemical Genomics Center [NCGC]
NIH Molecular Libraries Probe Centers Network [MLPCN]
MLPCN Grant: MH086850-01A1
Assay Submitter (PI): Robert Davey, University of Texas Medical Branch, Galveston, TX
For screening, 1000 cells in 2 ul/well were dispensed into white solid 1536-well plates (Greiner) using a solenoid-based dispenser. Following transfer of 23nl compound or DMSO vehicle by a pin tool, 3 ul/well of VSV-LV was added. The plates were centrifuged 1 min at 1000 RPM and then incubated 16 hr at 37 C and 5% CO2. After addition of 4 ul/well SteadyLite (PerkinElmer) detection reagent, the plates were incubated 10 min at ambient temperature and luminescence was measured on a ViewLux (Perkin Elmer) plate reader.
Keywords: NIH Roadmap, MLPCN, MLI, MLSMR, qHTS, NCGC, Lassa virus, luciferase, cell assay, infection
1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent inhibitors are ranked higher than compounds that showed apparent activation.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.
Categorized Comment - additional comments and annotations
* Activity Concentration. ** Test Concentration.
Data Table (Concise)