Location: Foreign Animal Disease Research
Project Number: 1940-32000-055-00
Start Date: Mar 12, 2008
End Date: Mar 30, 2011
ARS:1.Determination of FMDV interference with host responses will be completed by a)examining the interference with the innate response in FMDV-infected cells, b)examining the mechanisms of FMDV immune evasion.2.Development of a novel live-attenuated CSF marker vaccine requires knowledge of the genetic basis of viral virulence.We will systematically evaluate the role of specific viral proteins or virus virulence & use this information to engineer recombinant LAVs.Introduction of modifications in CSFV infectious clone.UofMO:1.Knockout pigs will be generated containing specific deletions of genes of interest for pathogenesis & innate response.Target genes include the receptor for IFN alpha & beta;B2M or CD8alpha gene & IFN gamma receptor.Swine will be assessed for ability to respond to FMDV & CSF infections and vaccines or biotherapeutics.2.Utilizing cytopathic & non-sytopathic strains of bovine viral diarrhea virus as a model,will determine type I interferon activation pathways in cattle.3.Effects of Si RNAs targeting PKR & TLR3 on IFN induction will be determined.The role of suppressors of cytokine signaling in blocking IFN mediated IFN production.Identify viral genes that target innate response.Results will be contrasted with those obtained with CSF virus in swine.3.Utilizing cloned & expressed RNA-dependent-RNA polymerases(RdRps)from FMDV,BVDV & CSFV identify structural similarities & potential active sites.Identify potential compounds that can block enzyme activity & viral replication.Candidate inhibitors will be validated utilizing gel-based biochemical assays & high throughput surface plasmon resonance analysis,mass spectrometry &proteomics approaches.UofCT:1.Evaluation of mucosal adjuvants efficacy delivered through the ad5 platform,to induce mucosal immune responses to FMDV.Evaluate in vivo mucosal adjuvants alone or in combination with FMDV vaccine for induction of rapid protection in swine.Determine cross-neutralization & cross-protection provided by capsid-based vaccines engineered with chimeric VP1 G-H loops containing immunogenic or toleragenic epitopes.Epitope map will be created using anti-sera from murine bearing cross-reactive immune responses between FMDV types O & SAT3.Testing chimeric GH loop bearing hAd5 vectors in swine will be conducted to assess cross-neutralization.Challenge studies will be performed utilizing homo-typic & heterotypic virus.2.Evaluate the role of non-structural proteins in CSFV virulence & protection against infection will be performed through;complete cloning of CSFV structural proteins into Baculovirus transfer vectors, completing the production of recombinant Baculovirus expressing parental CSFV structural proteins & of autonomous replication CSFV defective genomes, & by completing the immunogenicity studies in naïve swine with sera from infected swine.3.Development & validation of GCSPRI device will be done to use as rapid detection of FMDV & CSFV.Identify diagnostic reagents & develop host immune response charactacterization.Assay conditions & sensor chip configurations will be optimized to capture host leukocyte populations.In vivo virus detection will be tested through immune response by GCSPRI.