title: Replication-competent fluorescent-expressing influenza B virus.
abstract: Influenza B viruses (IBVs) cause annual outbreaks of respiratory illness in humans and are increasingly recognized as a major cause of influenza-associated morbidity and mortality. Studying influenza viruses requires the use of secondary methodologies to identify virus-infected cells. To this end, replication-competent influenza A viruses (IAVs) expressing easily traceable fluorescent proteins have been recently developed. In contrast, similar approaches for IBV are mostly lacking. In this report, we describe the generation and characterization of replication-competent influenza B/Brisbane/60/2008 viruses expressing fluorescent mCherry or GFP fused to the C-terminal of the viral non-structural 1 (NS1) protein. Fluorescent-expressing IBVs display similar growth kinetics and plaque phenotype to wild-type IBV, while fluorescent protein expression allows for the easy identification of virus-infected cells. Without the need of secondary approaches to monitor viral infection, fluorescent-expressing IBVs represent an ideal approach to study the biology of IBV and an excellent platform for the rapid identification and characterization of antiviral therapeutics or neutralizing antibodies using high-throughput screening approaches. Lastly, fluorescent-expressing IBVs can be combined with the recently described reporter-expressing IAVs for the identification of novel therapeutics to combat these two important human respiratory pathogens.
authors.affiliation: Department of Microbiology and Immunology, University of Rochester, Rochester, NY 14642, USA.
authors.forename: Aitor
authors.initials: A
authors.lastname: Nogales
authors.affiliation: Department of Microbiology and Immunology, University of Rochester, Rochester, NY 14642, USA.
authors.forename: Irene
authors.initials: I
authors.lastname: Rodríguez-Sánchez
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA; Department of Medicine, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Kristen
authors.initials: K
authors.lastname: Monte
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA; Department of Medicine, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Deborah J
authors.initials: DJ
authors.lastname: Lenschow
authors.affiliation: Department of Population Health, Poultry Diagnostic and Research Center, University of Georgia, Athens, GA 30603, USA.
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Perez
authors.affiliation: Department of Microbiology and Immunology, University of Rochester, Rochester, NY 14642, USA. Electronic address: luis_martinez@urmc.rochester.edu.
authors.forename: Luis
authors.initials: L
authors.lastname: Martínez-Sobrido
mesh_terms: D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D009981:Influenza B virus / Q000235:genetics / Q000502:physiology*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods*; D017361:Viral Nonstructural Proteins / Q000235:genetics; D010948:Viral Plaque Assay; D014773:Virology / Q000379:methods; D014779:Virus Replication; D000097573:Red Fluorescent Protein
chemical_list: D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D017361:Viral Nonstructural Proteins; D000097573:Red Fluorescent Protein; C085820:INS1 protein, influenza virus
keywords: Antivirals; Fluorescent-based Microneutralization assays; GFP; Influenza B virus; Neutralizing antibodies; mCherry