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

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....

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Autores principales: Caì, Yíngyún, Iwasaki, Masaharu, Beitzel, Brett F., Yú, Shuīqìng, Postnikova, Elena N., Cubitt, Beatrice, DeWald, Lisa Evans, Radoshitzky, Sheli R., Bollinger, Laura, Jahrling, Peter B., Palacios, Gustavo F., de la Torre, Juan C., Kuhn, Jens H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6266387/
https://www.ncbi.nlm.nih.gov/pubmed/30463334
http://dx.doi.org/10.3390/v10110655
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author Caì, Yíngyún
Iwasaki, Masaharu
Beitzel, Brett F.
Yú, Shuīqìng
Postnikova, Elena N.
Cubitt, Beatrice
DeWald, Lisa Evans
Radoshitzky, Sheli R.
Bollinger, Laura
Jahrling, Peter B.
Palacios, Gustavo F.
de la Torre, Juan C.
Kuhn, Jens H.
author_facet Caì, Yíngyún
Iwasaki, Masaharu
Beitzel, Brett F.
Yú, Shuīqìng
Postnikova, Elena N.
Cubitt, Beatrice
DeWald, Lisa Evans
Radoshitzky, Sheli R.
Bollinger, Laura
Jahrling, Peter B.
Palacios, Gustavo F.
de la Torre, Juan C.
Kuhn, Jens H.
author_sort Caì, Yíngyún
collection PubMed
description 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.
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spelling pubmed-62663872018-12-07 Recombinant Lassa Virus Expressing Green Fluorescent Protein as a Tool for High-Throughput Drug Screens and Neutralizing Antibody Assays Caì, Yíngyún Iwasaki, Masaharu Beitzel, Brett F. Yú, Shuīqìng Postnikova, Elena N. Cubitt, Beatrice DeWald, Lisa Evans Radoshitzky, Sheli R. Bollinger, Laura Jahrling, Peter B. Palacios, Gustavo F. de la Torre, Juan C. Kuhn, Jens H. Viruses Article 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. MDPI 2018-11-20 /pmc/articles/PMC6266387/ /pubmed/30463334 http://dx.doi.org/10.3390/v10110655 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Caì, Yíngyún
Iwasaki, Masaharu
Beitzel, Brett F.
Yú, Shuīqìng
Postnikova, Elena N.
Cubitt, Beatrice
DeWald, Lisa Evans
Radoshitzky, Sheli R.
Bollinger, Laura
Jahrling, Peter B.
Palacios, Gustavo F.
de la Torre, Juan C.
Kuhn, Jens H.
Recombinant Lassa Virus Expressing Green Fluorescent Protein as a Tool for High-Throughput Drug Screens and Neutralizing Antibody Assays
title Recombinant Lassa Virus Expressing Green Fluorescent Protein as a Tool for High-Throughput Drug Screens and Neutralizing Antibody Assays
title_full Recombinant Lassa Virus Expressing Green Fluorescent Protein as a Tool for High-Throughput Drug Screens and Neutralizing Antibody Assays
title_fullStr Recombinant Lassa Virus Expressing Green Fluorescent Protein as a Tool for High-Throughput Drug Screens and Neutralizing Antibody Assays
title_full_unstemmed Recombinant Lassa Virus Expressing Green Fluorescent Protein as a Tool for High-Throughput Drug Screens and Neutralizing Antibody Assays
title_short Recombinant Lassa Virus Expressing Green Fluorescent Protein as a Tool for High-Throughput Drug Screens and Neutralizing Antibody Assays
title_sort recombinant lassa virus expressing green fluorescent protein as a tool for high-throughput drug screens and neutralizing antibody assays
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6266387/
https://www.ncbi.nlm.nih.gov/pubmed/30463334
http://dx.doi.org/10.3390/v10110655
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