title: Inhibiting avian influenza virus shedding using a novel RNAi antiviral vector technology: proof of concept in an avian cell model.
abstract: Influenza A viruses pose significant health and economic threats to humans and animals. Outbreaks of avian influenza virus (AIV) are a liability to the poultry industry and increase the risk for transmission to humans. There are limitations to using the AIV vaccine in poultry, creating barriers to controlling outbreaks and a need for alternative effective control measures. Application of RNA interference (RNAi) techniques hold potential; however, the delivery of RNAi-mediating agents is a well-known obstacle to harnessing its clinical application. We introduce a novel antiviral approach using bacterial vectors that target avian mucosal epithelial cells and deliver (small interfering RNA) siRNAs against two AIV genes, nucleoprotein (NP) and polymerase acidic protein (PA). Using a red fluorescent reporter, we first demonstrated vector delivery and intracellular expression in avian epithelial cells. Subsequently, we demonstrated significant reductions in AIV shedding when applying these anti-AIV vectors prophylactically. These antiviral vectors provided up to a 10,000-fold reduction in viral titers shed, demonstrating in vitro proof-of-concept for using these novel anti-AIV vectors to inhibit AIV shedding. Our results indicate this siRNA vector technology could represent a scalable and clinically applicable antiviral technology for avian and human influenza and a prototype for RNAi-based vectors against other viruses.
authors.affiliation: Animal Population Health Institute, Colorado State University, Fort Collins, CO, USA. Lyndsey.Linke@colostate.edu.|Department of Clinical Sciences, Colorado State University, Fort Collins, CO, USA. Lyndsey.Linke@colostate.edu.
authors.forename: Lyndsey M
authors.initials: LM
authors.lastname: Linke
authors.affiliation: Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO, USA.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Wilusz
authors.affiliation: Animal Population Health Institute, Colorado State University, Fort Collins, CO, USA.|Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO, USA.
authors.forename: Kristy L
authors.initials: KL
authors.lastname: Pabilonia
authors.affiliation: Cambridge Biolabs, Cambridge, MA, USA.
authors.forename: Johannes
authors.initials: J
authors.lastname: Fruehauf
authors.affiliation: Animal Population Health Institute, Colorado State University, Fort Collins, CO, USA.|Department of Clinical Sciences, Colorado State University, Fort Collins, CO, USA.
authors.forename: Roberta
authors.initials: R
authors.lastname: Magnuson
authors.affiliation: Department of Clinical Sciences, Colorado State University, Fort Collins, CO, USA.
authors.forename: Francisco
authors.initials: F
authors.lastname: Olea-Popelka
authors.affiliation: Animal Population Health Institute, Colorado State University, Fort Collins, CO, USA.|Department of Clinical Sciences, Colorado State University, Fort Collins, CO, USA.|Environmental Health Services, Colorado State University, Fort Collins, CO, USA.
authors.forename: Joni
authors.initials: J
authors.lastname: Triantis
authors.affiliation: Department of Clinical Sciences, Colorado State University, Fort Collins, CO, USA.
authors.forename: Gabriele
authors.initials: G
authors.lastname: Landolt
authors.affiliation: Animal Population Health Institute, Colorado State University, Fort Collins, CO, USA.|Department of Clinical Sciences, Colorado State University, Fort Collins, CO, USA.
authors.forename: Mo
authors.initials: M
authors.lastname: Salman
keywords: Avian influenza; Bacterial vector; Influenza antiviral; Transkingdom RNAi; siRNA delivery