Recombinant Lassa Virus Expressing Green Fluorescent Protein as a Tool for High-Throughput Drug Screens and Neutralizing Antibody Assays.
Viruses
; 10(11)2018 11 20.
Article
em En
| MEDLINE
| ID: mdl-30463334
ABSTRACT
Lassa virus (LASV), a mammarenavirus, infects an estimated 100,000â»300,000 individuals yearly in western Africa and frequently causes lethal disease. Currently, no LASV-specific antivirals or vaccines are commercially available for prevention or treatment of Lassa fever, the disease caused by LASV. The development of medical countermeasure screening platforms is a crucial step to yield licensable products. Using reverse genetics, we generated a recombinant wild-type LASV (rLASV-WT) and a modified version thereof encoding a cleavable green fluorescent protein (GFP) as a reporter for rapid and quantitative detection of infection (rLASV-GFP). Both rLASV-WT and wild-type LASV exhibited similar growth kinetics in cultured cells, whereas growth of rLASV-GFP was slightly impaired. GFP reporter expression by rLASV-GFP remained stable over several serial passages in Vero cells. Using two well-characterized broad-spectrum antivirals known to inhibit LASV infection, favipiravir and ribavirin, we demonstrate that rLASV-GFP is a suitable screening tool for the identification of LASV infection inhibitors. Building on these findings, we established a rLASV-GFP-based high-throughput drug discovery screen and an rLASV-GFP-based antibody neutralization assay. Both platforms, now available as a standard tool at the IRF-Frederick (an international resource), will accelerate anti-LASV medical countermeasure discovery and reduce costs of antiviral screens in maximum containment laboratories.
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Coloração e Rotulagem
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Testes de Neutralização
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Genes Reporter
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Proteínas de Fluorescência Verde
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Substâncias Luminescentes
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Avaliação Pré-Clínica de Medicamentos
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Vírus Lassa
Tipo de estudo:
Prognostic_studies
Limite:
Animals
Idioma:
En
Revista:
Viruses
Ano de publicação:
2018
Tipo de documento:
Article
País de afiliação:
Estados Unidos