Location: Animal Parasitic Diseases Laboratory
Project Number: 8042-32000-102-21-I
Project Type: Interagency Reimbursable Agreement
Start Date: Feb 15, 2015
End Date: Feb 14, 2018
Current animal health and biomedical research efforts necessitate use of a broad range of immune reagents, but those available for pigs are limited. Our goal is to generate priority immune tools for the pig veterinary and biomedical research community. The project team will prioritize research targets, seeking input from swine immunologists in the US, UK and worldwide. Our aim is to identify targets based on newly revealed immune pathways, gaps based on updated listing of commercially available reagents, and prevent duplication of effort. Based on experience gained from the US Veterinary Immune Reagent Network (VIRN) the team will identify best practices, protocols and screening methods, team with commercial experts for protein expression and monoclonal antibody (mAb) production, and evaluate targets for potential success based on results for human and rodent reagents. Specific Objectives: 1) Clone and express swine immune cytokines and chemokines; 2) Prepare panels of mAb reactive with swine targets; 3) Use reagents produced to develop new assays for swine immune markers; and 4) Provide the veterinary community with new commercial reagents and up-to-date information and techniques for their research efforts.
For Objective 1 Develop immunologic tools to evaluate swine immunity, including using immunological tools to enhance our understanding of swine immune system development [C4, PS4B], and using immunological tools to inform the design of novel innate immune intervention strategies to treat respiratory diseases of swine. For Objective 2 panels of monoclonal antibodies (mAbs) will be produced under contract for selected cell surface or CD markers and swine cytokines, chemokines and receptors. This process can start immediately as there are already expressed cytokines (e.g., IL-17A) at Kingfisher Biotech, Inc. for which no panel of mAb is yet available. Each immunization and fusion is expected to result in a panel of hybridoma clones for each target. These will enable the team to identify epitope reactivity and utility for different assays (verified in Objective 3) and use advanced UK immunofluorescence screening techniques to identify molecular interactions in tissues. For Objective 3 new assays for swine immune markers will be established. These will include identifying mAb pairs that work for ELISA and multiplex assays for cytokines and chemokines. The National Pork Board funded the development of the first multiplex assays for cytokines. Other targets will be mAb reactive for intercellular staining or for cell receptor binding/blocking. For Objective 4 there will be commercial sources for reagents and updates to the community. Kingfisher Biotech, Inc. will continue to market expressed cytokines and chemokines; each hybridoma producer will serve as a source for mAb products. Information will be disseminated to swine researchers via regular updates, e.g., posters at relevant meetings as well as publications.