High-throughput screening for broad-spectrum chemical inhibitors of RNA viruses.
RNA viruses are responsible for major human diseases such as flu, bronchitis, dengue, Hepatitis C or measles. They also represent an emerging threat because of increased worldwide exchanges and human populations penetrating more and more natural ecosystems. A good example of such an emerging situation is chikungunya virus epidemics of 2005-2006 in the Indian Ocean. Recent progresses in our understanding of cellular pathways controlling viral replication suggest that compounds targeting host cell functions, rather than the virus itself, could inhibit a large panel of RNA viruses. Some broad-spectrum antiviral compounds have been identified with host target-oriented assays. However, measuring the inhibition of viral replication in cell cultures using reduction of cytopathic effects as a readout still represents a paramount screening strategy. Such functional screens have been greatly improved by the development of recombinant viruses expressing reporter enzymes capable of bioluminescence such as luciferase. In the present report, we detail a high-throughput screening pipeline, which combines recombinant measles and chikungunya viruses with cellular viability assays, to identify compounds with a broad-spectrum antiviral profile.
Unité de Génomique Virale et Vaccination, Virology Department, Institut Pasteur, CNRS UMR3569.
Marianne
M
Lucas-Hourani
Unité de Chimie et Biocatalyse, Biochemistry and Structural Biology Department, Institut Pasteur, CNRS UMR3523.
Hélène
H
Munier-Lehmann
Unité de Chimie et Biocatalyse, Biochemistry and Structural Biology Department, Institut Pasteur, CNRS UMR3523.
Olivier
O
Helynck
Unité de Génomique Virale et Vaccination, Virology Department, Institut Pasteur, CNRS UMR3569.
Anastassia
A
Komarova
Unité des Interactions Moléculaires Flavivirus-Hôtes, Virology Department, Institut Pasteur.
Philippe
P
Desprès
Unité de Génomique Virale et Vaccination, Virology Department, Institut Pasteur, CNRS UMR3569.
Frédéric
F
Tangy
Unité de Génomique Virale et Vaccination, Virology Department, Institut Pasteur, CNRS UMR3569; vidalain@pasteur.fr.
Pierre-Olivier
PO
Vidalain
D000998:Antiviral Agents / Q000494:pharmacology*; D002646:Chikungunya virus / Q000187:drug effects* / Q000235:genetics; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D049409:Luciferases, Firefly / Q000096:biosynthesis / Q000235:genetics; D008459:Measles virus / Q000187:drug effects* / Q000235:genetics
D000998:Antiviral Agents; D049409:Luciferases, Firefly