Identification of avian RIG-I responsive genes during influenza infection

• Published in: Molecular immunology 2013-05, Vol.54 (1), p.89-97
• Main Authors: Barber, Megan R.W., Aldridge, Jerry R., Fleming-Canepa, Ximena, Wang, Yong-Dong, Webster, Robert G., Magor, Katharine E.
• Format: Article
• Language: English
• Subjects: Animals; Birds - genetics; Birds - immunology; Cells, Cultured; Chick Embryo; Chickens - genetics; Chickens - immunology; Cluster Analysis; DEAD-box RNA Helicases - physiology; Ducks - genetics; Ducks - immunology; Gene Expression Profiling; H5N1; IFIT; Immunity, Innate - genetics; Immunity, Innate - immunology; Influenza A; Influenza in Birds - genetics; Influenza in Birds - immunology; Interferon stimulated genes; Microarray; Microarray Analysis; MX1; OASL; PKR; RIG-I
• ISSN: 0161-5890; 1872-9142
• DOI: 10.1016/j.molimm.2012.10.038

► 44K Agilent microarray analyses of chicken DF-1 cells with duck RIG-I or empty vector. ► Transcriptome of H5N2 infected cells with duck RIG-I or empty vector. ► Transcriptome of H5N1 infected cells with duck RIG-I or empty vector. ► Identification of genes augmented in expression by duck RIG-I. ► Quantitative real-time PCR confirms upregulation of key innate immune genes. Ducks can survive infection with highly pathogenic avian influenza viruses that are lethal to chickens. We showed that the influenza detector, RIG-I can initiate antiviral responses in ducks, but this gene is absent in chickens. We can reconstitute this pathway by transfecting chicken DF-1 embryonic fibroblast cells with duck RIG-I, which augments their antiviral response to influenza and decreases viral titer. However, the genes downstream of duck RIG-I that mediate this antiviral response to influenza are not known. Using microarrays, we compared the transcriptional profile of chicken embryonic fibroblasts transfected with duck RIG-I or empty vector, and infected with low or highly pathogenic avian influenza viruses. Transfected duck RIG-I expressed in chicken cells was associated with the marked induction of many antiviral innate immune genes upon infection with both viruses. We used real-time PCR to confirm upregulation of a subset of these antiviral genes including MX1, PKR, IFIT5, OASL, IFNB, and downregulation of the influenza matrix gene. These results provide some insight into the genes induced by duck RIG-I upon influenza infection, and provide evidence that duck RIG-I can function to elicit an interferon-driven, antiviral response against influenza in chicken embryonic fibroblasts.