Recombinant Lassa Virus Expressing Green Fluorescent Protein as a Tool for High-Throughput Drug Screens and Neutralizing Antibody Assays

Author:

Caì Yíngyún,Iwasaki Masaharu,Beitzel Brett,Yú Shuīqìng,Postnikova Elena,Cubitt Beatrice,DeWald Lisa,Radoshitzky Sheli,Bollinger Laura,Jahrling Peter,Palacios Gustavo,de la Torre Juan,Kuhn Jens

Abstract

Lassa virus (LASV), a mammarenavirus, infects an estimated 100,000–300,000 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.

Funder

National Institute of Allergy and Infectious Diseases

Publisher

MDPI AG

Subject

Virology,Infectious Diseases

Reference49 articles.

1. World Health Organization (WHO), List of Blueprint Priority Diseases http://www.who.int/blueprint/priority-diseases/en/

2. Lassa fever, a new virus disease of man from West Africa. I. Clinical description and pathological findings;Frame;Am. J. Trop. Med. Hyg.

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