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27. Quantifying the interferon-stimulated immune response after MAVS cleavage by Coxsackievirus B3 (CVB3) proteases 2Apro and 3Cpro expressed in cardiomyocytes

Presenters Name: 
Page Murray
Co Presenters Name: 
Primary Research Mentor: 
Kevin Janes
Secondary Research Mentor: 
Andrew Sweatt
Session: 
1
Grant Program Recipient: 
Not a Recipient
Abstract: 

Enteroviruses cause or are associated with many health complications such as myocarditis and dilated cardiomyopathy. In enterovirus infections, viral double-stranded RNA stimulates Mitochondrial Antiviral Signaling protein (MAVS). MAVS activation results in expression of interferons and interferon stimulated genes (ISGs), which have antiviral properties. Enteroviral proteases 2Apro and 3Cpro cleave MAVS to inhibit this antiviral effect. The roles of 2Apro and 3Cpro in the immune response remain unclear because it is difficult to separate the effects of the viral proteins. In this study, we use Coxsackievirus B3 as a model enterovirus and quantify the effects of 2Apro and 3Cpro on MAVS, interferon transcription factors (NF-kB and IRF3) and ISGs  (ISG15, OAS1, OAS2). To isolate the proteases‚Äô effects, we established cardiomyocyte cell lines to inducibly express either 2Apro or 3Cpro. We have confirmed 3Cpro expression, and 2Apro cells are undergoing selection. In cells expressing 2Apro or 3Cpro, the MAVS pathway will be activated by Poly(I:C), which mimics double-stranded RNA. The cells will be lysed for protein and RNA. Immunoblotting and qPCR will be used to quantify changes in production of interferon transcription factors and ISGs. We hypothesize that 3Cpro will more efficiently cleave MAVS, suppress interferon production, and inactivate the innate immune response. Therefore, we expect to see abundant cell death or injury after protease expression. Clinically, larger amounts of cell death lead to severe health complications.  My research identifies functional differences of each protease, a critical step in developing novel therapies against enteroviruses.