title: Functional characterization of canine interferon-lambda.
abstract: In this study, we provide the first comprehensive annotation of canine interferon-λ (CaIFN-λ, type III IFN). Phylogenetic analysis based on genomic sequences indicated that CaIFN-λ is located in the same branch with Swine IFN-λ1 (SwIFN-λ), Bat IFN-λ1 (BaIFN-λ), and human IFN-λ1 (HuIFN-λ1). CaIFN-λ was cloned, expressed in Escherichia coli, and purified to further investigate the biological activity in vitro. The recombinant CaIFN-λ (rCaIFN-λ) displayed potent antiviral activity on both homologous and heterologous animal cells in terms of inhibiting the replication of the New Jersey serotype of vesicular stomatitis virus (VSV), canine parvovirus, and influenza virus A/WSN/33 (H1N1), respectively. In addition, we also found that rCaIFN-λ exhibits a significant antiproliferative response against A72 canine tumor cells and MDCK cells in a dose-dependent manner. Furthermore, CaIFN-λ activated the JAK-STAT signaling pathway. To evaluate the expression of CaIFN-λ induced by virus and the expression of IFN-stimulated genes (ISGs) induced by rCaIFN-λ in the MDCK cells, we measured the relative mRNA level of CaIFN-λ and ISGs (ISG15, Mx1, and 2'5'-OAS) by quantitative real-time PCR and found that the mRNA level of CaIFN-λ and the ISGs significantly increased after treating the MDCK cells with viruses and rCaIFN-λ protein, respectively. Finally, to evaluate the binding activity of rCaIFN-λ to its receptor, we expressed the extracellular domain of the canine IFN-λ receptor 1 (CaIFN-λR1-EC) and determined the binding activity via ELISA. Our results demonstrated that rCaIFN-λ bound tightly to recombinant CaIFN-λR1-EC (rCaIFN-λR1-EC).
authors.affiliation: 1 College of Animal Sciences and Veterinary Medicine, Guangxi University , Nanning, People's Republic of China .
authors.forename: Wenhui
authors.initials: W
authors.lastname: Fan
authors.forename: Lei
authors.initials: L
authors.lastname: Xu
authors.forename: Liqian
authors.initials: L
authors.lastname: Ren
authors.forename: Hongren
authors.initials: H
authors.lastname: Qu
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.forename: Jingjing
authors.initials: J
authors.lastname: Liang
authors.forename: Wenjun
authors.initials: W
authors.lastname: Liu
authors.forename: Limin
authors.initials: L
authors.lastname: Yang
authors.forename: Tingrong
authors.initials: T
authors.lastname: Luo
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D004285:Dogs; D006131:Growth Inhibitors / Q000378:metabolism*; D006801:Humans; D007372:Interferons / Q000145:classification* / Q000235:genetics / Q000276:immunology*; D007378:Interleukins / Q000145:classification* / Q000235:genetics / Q000276:immunology*; D053612:Janus Kinases / Q000378:metabolism; D061985:Madin Darby Canine Kidney Cells; D008969:Molecular Sequence Data; D010802:Phylogeny; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D050791:STAT Transcription Factors / Q000378:metabolism; D015398:Signal Transduction; D014777:Virus Diseases / Q000276:immunology; D014779:Virus Replication
chemical_list: D006131:Growth Inhibitors; D007378:Interleukins; D011994:Recombinant Proteins; D050791:STAT Transcription Factors; D007372:Interferons; D053612:Janus Kinases