title: Antisense oligonucleotide inhibits avian influenza virus H5N1 replication by single chain antibody delivery system.
abstract: H5N1 avian influenza virus (AIV) causes widespread infections in poultry and wild birds, and has the potential to emerge as a pandemic threat to human. Antisense oligonucleotides (AS ODNs) are highly effective at inhibiting gene replication. Antibody-mediated delivery is a novel approach to target specific cells and tissues. In this study, we designed and synthesized three AS ODNs (PA4, PA492 and PA1203) specific for conserved region of AIV PA protein, and all the three AS ODNs could inhibit viral replication. The PA492 ODN showed the best antiviral effect by viral titers and quantitative RT-PCR in MDCK cells. The fusion protein scFv-tP was constructed as a single chain variable fragment (scFv) against AIV hemaglutinin antigen with a truncated protamine (tP). The results showed that scFv-tP fusion improved the antiviral effectiveness of PA492 in MDCK cells as measured by viral titers, quantitative RT-PCR and indirect immunofluorescence (IFA) assays. In addition, scFv-tP-delivered PA492 was also found to partially protect mice from lethal H5N1 influenza virus challenge. Using scFv-tP delivery, fluorescein isothiocyanate labeled-PA492 was found to be significantly localized in the lungs, compared to liposome-delivered PA492. Moreover, the fusion protein mediated PA492 had a lower lung index and viral titers in the infected mice as compared with the liposome method. These results provided a potential method for using anti-HA fusion protein for the targeted delivery of AS ODNs against AIV H5N1.
authors.affiliation: Institute of Laboratory Animal Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100021, China. zhangtao1012450@gmail.com
authors.forename: Tao
authors.initials: T
authors.lastname: Zhang
authors.forename: Ping-sen
authors.initials: PS
authors.lastname: Zhao
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Meng
authors.initials: M
authors.lastname: Liang
authors.forename: Yu-wei
authors.initials: YW
authors.lastname: Gao
authors.forename: Song-tao
authors.initials: ST
authors.lastname: Yang
authors.forename: Tie-cheng
authors.initials: TC
authors.lastname: Wang
authors.forename: Chuan
authors.initials: C
authors.lastname: Qin
authors.forename: Cheng-yu
authors.initials: CY
authors.lastname: Wang
authors.forename: Xian-zhu
authors.initials: XZ
authors.lastname: Xia
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D002645:Chickens; D004285:Dogs; D016503:Drug Delivery Systems; D005260:Female; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology; D053124:Influenza A Virus, H5N1 Subtype / Q000502:physiology*; D008081:Liposomes / Q000494:pharmacology; D008168:Lung / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D011993:Recombinant Fusion Proteins / Q000494:pharmacology; D057127:Single-Chain Antibodies / Q000494:pharmacology*; D019562:Viral Load; D014779:Virus Replication
chemical_list: D000998:Antiviral Agents; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D008081:Liposomes; D016376:Oligonucleotides, Antisense; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies