1. Characterization of the antiviral properties of IFIT3: Recently, we have further elucidated the antiviral role of IFIT3 by showing that overexpression of IFIT3 decreases Dengue Virus replication in HEK293 cells. 2. The Role of Ly6E in eliciting an antiviral effect: Through the modeling and sequence alignment we created a rabbit monoclonal antibody to a unique region of Ly6E. We validated the antibody both through Western Blot (WB) and Flow Cytometric (FC) analysis. Using FC and WB we found that A549 (human lung adenocarcinoma) cells expressed Ly6E constitutively, independent of IFN-. Furthermore, basic biochemical analysis has found that the protein is highly unstable in a wide variety of storage conditions, and cannot be recovered readily after being dissociated from the cell, indicating that it has unique biochemical properties. 3. The Differential Regulation of Type I Interferons in Response to Viral Infection: We analyzed the interferon signaling mediators in several different cell lines that were infected with 3 different viruses in order to gain a better understanding in the signaling pathways involved in the induction of interferons and interferon stimulated genes. This information can then be used to manipulate the interferon response to preferentially induce targets that have greater antiviral potencies. 4. Enhancement of the antiviral activity of IFN- by Ribavirin: Combination treatments of interferon and ribavirin are successful in treating viral infections, however the mechanism of action for how these agents work together remains unclear. To examine this, we utilized vesicular stomatitis virus infection on A549 human lung epithelial cells as an experimental model. Treating cells with ribavirin alone or in combination with interferon resulted in increased and prolonged activation of pSTAT1 and pSTAT2, and in increases in total STAT1, STAT2, and IRF9 protein expression compared to interferon alone. In addition, expression of the Interferon Stimulated Genes (ISGs) MxA, IFIT3, PKR, and ISG15 showed enhanced expression in the presence of ribavirin alone or in combination with interferon when compared to interferon alone. This prolonged and enhanced activity of interferon signaling and induction of gene expression may play a role in how interferon and ribavirin work together to treat viral infections.

Project Start
Project End
Budget Start
Budget End
Support Year
6
Fiscal Year
2013
Total Cost
$535,849
Indirect Cost
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