Progress 07/20/04 to 07/19/09
Outputs
Progress Report Objectives (from AD-416) Develop novel strategies to reduce economic losses due to intestinal coccidia parasites: define mucosal cell-mediated immune mechanisms which confer host protection against coccidiosis; identify chicken cytokines which confer immune enhancement against parasites using high throughput gene screening; and identify potential vaccine antigens for Eimeria acervulina and develop viral vectors and direct DNA injection systems for coccidiosis vaccines. Approach (from AD-416) Define mucosal cell-mediated immune mechanisms which confer host protection against coccidia. Clone and express various recombinant host genes, prepare monoclonal antibodies and sensitive immunological and molecular techniques to assess avian immune responses to pathogens and to envaluate local cell-mediated immunity, identify chicken cytokines which enhance responses against parasites using high throughput gene screening systems, the identify of host genes which enhance host protective immune responses will be identified, identify potential vaccine antigens for Eimeria acervulina; develop viral vectors; direct DNA injection systems. DNA from Eimeria spp., will be introduced into vaccine strains of viruses commonly used in the poultry industry. The first round of vaccine trials will entail immunization of chickens at one day of age. If protective efficacy is achieved, then in ovo injection of recombinant vaccines will proceed. Significant Activities that Support Special Target Populations This is a final report for this agreement. This is a CRADA developed with the Diversa company to develop recombinant coccidiosis vaccine strategy and to discover novel host genes which encode defensins. ARS developed in vivo and in vitro assays to evaluate gene functions and vaccine efficacy in coccidiosis disease model and demonstrated immunogenicity of several new Eimeria genes from different developmental life stages.
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Progress 10/01/06 to 09/30/07
Outputs
Progress Report Objectives (from AD-416) Develop novel strategies to reduce economic losses due to intestinal coccidia parasites: define mucosal cell-mediated immune mechanisms which confer host protection against coccidiosis; identify chicken cytokines which confer immune enhancement against parasites using high throughput gene screening; and identify potential vaccine antigens for Eimeria acervulina and develop viral vectors and direct DNA injection systems for coccidiosis vaccines. Approach (from AD-416) Define mucosal cell-mediated immune mechanisms which confer host protection against coccidia. Clone and express various recombinant host genes, prepare monoclonal antibodies and sensitive immunological and molecular techniques to assess avian immune responses to pathogens and to envaluate local cell-mediated immunity, identify chicken cytokines which enhance responses against parasites using high throughput gene screening systems, the identify of host genes which enhance host protective immune responses will be identified, identify potential vaccine antigens for Eimeria acervulina; develop viral vectors; direct DNA injection systems. DNA from Eimeria spp., will be introduced into vaccine strains of viruses commonly used in the poultry industry. The first round of vaccine trials will entail immunization of chickens at one day of age. If protective efficacy is achieved, then in ovo injection of recombinant vaccines will proceed. Significant Activities that Support Special Target Populations This report documents research conducted under a Trust Agreement between ARS and Diversa Coorporation. Additional details of the research can be found in the report for the associated in-house project 1265-31320-072- 00D, �Development of Functional Genomic and Immunologic Approaches to Control Avian Mucosal Pathogens�. In this reporting period, we developed an experimental disease challenge model for avian coccidiosis to evaluate the immunoenhancing effects of potential vaccine candidate antigens of Eimeria. Especially, we have evaluated the vaccine potential of transhydrogenase (TH) and heat shock proteins (HSP). In two vaccine trials, we have vaccinated broiler chickens with two different recombinant proteins provided by Diversa. After vaccination, broiler chickens were challenged with live parasites and fecal oocysts production and body weight gains were evaluated. The level of protection induced by these recombinant proteins depended upon the dose and mode of immunization. Monitoring activity for this grant was in regular email and phone conversations with contacts at Diversa Corporation.
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Progress 10/01/05 to 09/30/06
Outputs
Progress Report 4d Progress report. This report serves to document research conducted under a CRADA between ARS and Diversa Corporation. Additional details of research can be found in the report for the parent project 1265-31320-070-00D:Genomic, Proteomic, and Immunologic Approaches to Controlling Avian Coccidiosis. This is a CRADA established with Diversa Incorporation in 2004 to investigate proprietary T-cell epitope vaccine developed by Diversa scientists. Recombinant coccidia vaccines expression 3-1E and MIC2 were produced using GMGT vector and tested by in ovo vaccination protocol. The results indicate that GMGT vectors enhance immunogenicity of 3-1E and MIC2 DNA vaccine. GMGT recombinant vaccines carrying HSP90 is being constructed and its expression level being evaluated.
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Progress 10/01/04 to 09/30/05
Outputs
4d Progress report. This report serves to document research conducted under a Trust Agreement between ARS and Avicore. Additional details of research can be found in the report for the parent project 1265-31320-070-00D: Genomic, Proteomic, and Immunologic Approaches to Controlling Avian Coccidiosis. Peroxiredoxin (PRX) is a crucial antioxidant protein that protects against endogenously produced peroxides in prokaryotes to eukaryotes. To date, six different isoforms have been identified in mammals. In this study, we describe the first members of the PRX protein family to be characterized in Aves. Through bioinformatics analysis, we observed that at least four different classes of PRX protein were evolutionarily conserved in Aves. Furthermore, in vitro functional assays of the candidate chicken PRX proteins demonstrated that they had similar levels of antioxidant activity to those of the mammalian enzymes. The expression patterns of the PRX transcript in several chicken
tissues were not tissue- specific, suggesting that they might play an essential role as a housekeeping gene in all tissues, to protect against oxidative damage. In conclusion, the sequences of the putative members of this functional gene family in chickens could be effectively retrieved in silico through bioinformatics analysis, and the functionality of their gene products evaluated by in vitro comparative assay.
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