1.We evaluated interferon and interferon stimulated gene production in response to cellular infection of different viruses in a variety of cell types in order to gain an understanding of the different signaling modulators that are activated when certain interferon subtypes are induced. We found that a group of subtypes are exclusively induced following stimulation of the IFN receptor, while another group of subtypes are exclusively induced following pattern recognition receptor stimulation. These groups of interferons are dependent on the cell type. We also found that distinct sets of interferon stimulated genes are induced depending on virus concentration. We also discovered that type I interferon pre-treatment leads to increased cytopathic effects and viral replication in certain cell types that are infected with Sendai virus even though the activation of pattern recognition receptors and JAK/STAT signalling modulators as well as the induction of interferon stimulated genes were observed. 2. We have shown that the uncharacterized protein FLJ11286 (FLJ) is induced by dengue virus infection in an IFN dependent manner and plays a role in antiviral response. FLJ is 291 aa in length, contains conserved cysteine residues and has homologues across the vertebrate taxon. FLJ is upregulated in response to stimulation with IFN, and that disruption of IFN signaling, through siRNA mediated knockdown and CRISPR Cas9 knockouts of IFN regulatory factor 9, results in decreased induction of FLJ after IFN treatment. FLJ displays antiviral activity against DENV as well as against Encephalomyocarditis virus. We also have demonstrated that FLJ is a nucleic acid binding protein with the ability to bind to dengue viral mRNA and associates with a variety of dengue and host proteins. These data serve to identify FLJ as a novel IFN stimulated gene with antiviral activity against DENV. 3.We also examined the role of IFIT3 in the antiviral response by studying the effect of IFIT3 on virus titers of VSV (vesicular stomatitis virus), EMCV (encephalomyocarditis virus) and DV (Dengue virus). In our studies, IFIT3 is the most up-regulated IFIT family member in response to treatment with interferon IFN-alpha 2a in certain cancer cell lines. Overexpression of IFIT3 demonstrates its antiviral activity in HEK293 cells against VSV and EMCV. IFIT3 may interact with IFIT1 and this interaction may play a role in regulation of protein synthesis and ISG expression. Although the precise mechanism is still unknown, IFIT3 demonstrated a clear antiviral effect against several different viruses. 4. Our ebola studies showed that although IFN can inhibit replication in vitro, it has very limited protection in nonhuman primates. 5. Like DENV, ZIKV has been shown to be sensitive to the effects of type I IFN. We have examining how ZIKV interacts with the host immune cells. CHanges in host gene expression after infection with ZIKV have been monitored by microarray analysis as well as RT-PCRT and western blot.
