2007 Annual Report
1a.Objectives (from AD-416)
This proposal presents a broad community plan to begin to systematically address the immunological reagent gap for the U.S. veterinary immunology research community including for the following groups: ruminants (concentrating on cattle), swine, poultry (primarily chickens with some evaluation of reagents on turkey cells), horses, and aquaculture species (concentrating on channel catfish and trout) with a goal of 20 reagents per species group. The reagents will include bioactive recombinant cytokines and chemokine proteins, expressed using mammalian, yeast, and bacterial systems, as well as antibodies to them and their receptors as well as antibodies to immunoglobulin isotypes, T cell receptors and other CD molecules. The goal is to produce antibodies that function in ELISA and ELISpot assays, for intracellular staining, for blocking function and signaling, and that are useful in flow cytometric applications as well as in fixed tissue sections. Antibodies to bioactive molecules will also be tested for their ability to block the recombinant molecules as well as natural molecules if it is possible to produce cell culture systems for the later. This project represents the swine component.
1b.Approach (from AD-416)
Prioritization of reagents (tools) to be produced will be established by surveying the larger scientific community and cooperating with other similar international efforts to avoid redundancy and target the most useful reagents for research scientists. Commercial companies will also be surveyed. Bioactive cytokines and chemokines and cell surface CD molecules, antibodies, and T cell receptors will be produced in expression systems in two main labs for poultry and mammals and by the two co-PDs for fish. The expression of these proteins will depend upon provision of cDNA from appropriate cells or tissues from the co-PD labs who are specialists in particular species. RT-PCR using primers based on available sequences or from homologous genes in other related species will be done to obtain the entire gene sequence. The recombinant molecules will be used to prime mice for production of monoclonal antibodies or rabbits for polyclonal antibodies in a centralized lab. After identifying reactive antibodies initially, both the recombinant molecules and the antibodies to them will be evaluated for efficacy in the species specialist labs in a variety of in vitro assays. This project will have oversight by an advisory board composed of industry partners, USDA national program directors and other international reagent-generating groups. The reagents will be provided to the scientific community by depositing in commercial cell banks and encouraging generation and distribution through commercial vendors.
This report documents research conducted under a Reimbursable Agreement between ARS and the Univ. of Massachusetts through USDA CSREES grant to Univ. Massachusetts, Amherst for the Veterinary Immunological Reagents Network with the USDA ARS BARC APDL scientist serving as the Chair for swine reagents. Additional details of research can be found in the report for the parent project 1265-32000-079-00D, "Strategies to Control Swine Parasites Affecting Food Safety."
A major obstacle to advances in veterinary immunology and disease control is the lack of sufficient immunological reagents. This USDA CSREES was funded in 2006 through a USDA CSREES grant to Univ. Massachusetts, Amherst, to establish the Veterinary Immunological Reagents Network. The Network was established to address the immunological reagent gap for the US veterinary immunology research community. This is 4 year multi-species effort supports cloning and expression of immunological reagents specific for ruminants, swine, poultry, equine and aquaculture species. The USDA ARS BARC APDL scientist serves as the Chair for swine reagents. The emphases for swine is on developing and characterizing bioactive immune proteins, cloned cytokines (interferons and interleukins) and chemokine proteins, as well as monoclonal antibodies (mAbs) to these proteins and their receptors. The prioritiy list is maintained on the network website http://www.umass.edu/vetimm/swine/index.html. Additional mAb will be characterized for toll-like receptor (TLR) proteins or produced to swine cell subset proteins, the CD antigens, and the T cell receptors. Reagents developed with this grant will be used for disease and vaccine research by veterinary immunologists, pathologists and microbiologists.
National Program 103 Component 2: Genetic and biological determinants of disease susceptibility
The ADODR for this grant helped to establish, and regularly participate in, monthly conference calls that review plans and progress for reagent development under this grant. Rregular email contact with the participating labs as they develop reagents were also exchanged.