The Cardiac Response to Reovirus Infection

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dc.contributor.advisor Barbara Sherry, Committee Chair en_US
dc.contributor.advisor Frank Scholle, Committee Member en_US
dc.contributor.advisor James L. Stephenson, Jr., Committee Member en_US
dc.contributor.advisor Jonathan M. Horowitz, Committee Member en_US
dc.contributor.author Li, Lianna en_US
dc.date.accessioned 2010-04-02T18:42:17Z
dc.date.available 2010-04-02T18:42:17Z
dc.date.issued 2009-08-03 en_US
dc.identifier.other etd-06302009-095014 en_US
dc.identifier.uri http://www.lib.ncsu.edu/resolver/1840.16/4015
dc.description.abstract LI, LIANNA. The Cardiac Response to Reovirus Infection. (Under the direction of Dr. Barbara Sherry). Viral myocarditis is a common disease in humans. Interferon-β (IFN-β) has been identified as critical for protection against viral myocarditis in mouse models, and IFN-α or -β treatment is beneficial in the treatment of human viral myocarditis. IFN-β expression and its antiviral effects are cell-type specific in murine cardiac myocytes and fibroblasts. However, expression and function of individual IFN-α subtypes in cardiac cells has not previously been investigated. Therefore, IFN-α subtype expression and antiviral effects were studied in reovirus-infected murine primary cardiac myocyte and cardiac fibroblast cultures. In order to quantify the thirteen highly conserved IFN-α subtype, a quantitative Real-Time PCR assay was developed. Results demonstrated that IFN-α induction by reovirus T3D in cardiac cells is both subtype- and cell type-specific, and that some individual IFN-α subtypes are likely important in the antiviral cardiac response. In brief, reovirus T3D induced five IFN-α subtypes in primary cultures of cardiac myocytes and fibroblasts: IFN-α1, -α2, -α4, -α5, and -α8/6. The levels of IFN-α expression were both higher and spanned a greater range in cardiac myocytes than in fibroblasts. Viral induction of IFN-α1, -α2, -α5, and -α8/6 required IFN-α/β signaling in both cell types, while induction of IFN-β and -α4 was more dependent on IFN signaling in myocytes than fibroblasts. Murine IFN-α1, -α2, -α4, or -α5 treatment induced IRF7 and ISG56 in both cardiac cell types, however induction was always greater in cardiac fibroblasts than in cardiac myocytes. Finally, each IFN-α subtype inhibited reovirus T3D replication in both cell types, but protection was subtype-specific. To discover novel proteins or protein post-translational modifications involved in the IFN pathway or displaying antiviral effects against viral myocarditis, a proteomics tool, two-dimensional difference gel electrophoresis (2D-DIGE) coupled with MALDI-TOF-TOF, was used to investigate the reovirus-induced proteome changes in murine primary cardiac myocyte cultures. Results demonstrated that the 2D-DIGE technique is quantitative and reproducible. Whole proteome changes based on differentially expressed proteins were clustered according to viral pathogenic phenotypes and induction of IFN. One hundred and twenty-four differentially expressed proteins were identified, including those involved in calcium signaling, ERK/ MAPK signaling, protein ubiquitination, mitochondrial dysfunction, oxidative stress, amino acid metabolism, and other pathways. Interestingly, 2D-DIGE results and additional studies demonstrated that heat shock protein Hsp25 is modulated differentially by myocarditic and non-myocarditic reoviruses, and suggested that it may play a role in the cardiac antiviral response. This is the eighth virus family found to modulate Hsp25 or its human homolog, Hsp27, suggesting that Hsp25/27 participation in the antiviral response may be widespread. However, results here provide the first evidence for a virus-induced decrease in Hsp25/27, and suggest that viruses may have evolved a mechanism to subvert this protective response, as they have for IFN. en_US
dc.rights I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dis sertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to NC State University or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. en_US
dc.subject Heart en_US
dc.subject Hsp25 en_US
dc.subject 2D-DIGE en_US
dc.subject Interferon en_US
dc.subject Myocarditis en_US
dc.subject Reovirus en_US
dc.subject Proteomics en_US
dc.title The Cardiac Response to Reovirus Infection en_US
dc.degree.name PhD en_US
dc.degree.level dissertation en_US
dc.degree.discipline Functional Genomics en_US


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