Submitted to: Veterinary Immunology and Immunopathology
Publication Type: Peer reviewed journal
Publication Acceptance Date: 1/30/2014
Publication Date: 2/28/2014
Citation: Lee, S.H., Lillehoj, H.S., Jeong, M., Cacho, E.D., Min, W., Sullivan, Y.B., Kakach, L., Labresh, J.W. 2014. Development and characterization of mouse monoclonal antibodies reactive with chicken TL1A. Veterinary Immunology and Immunopathology. 159(1-2):103-9. doi: 10.1016/j.vetimm.2014.01.002. Interpretive Summary: There is severe lack of reagents that scientists can use to evaluate the immune response of chickens in basic and applied research. This inability to evaluate poultry immunity hinders progress in understanding critical immune molecules of chickens that play important roles in promoting or resisting infections. In this study, ARS scientists collaborated with other scientists to identify a factor (gene) in chickens designated as TL1A which encodes the production of a protein that promotes inflammation. This TL1A protein was made in the laboratory to study its function. Furthermore, the TL1A protein was used to develop immune reagents which can detect this inflammatory protein in diseased chickens. Availability of these reagents will enhance scientists in industry and universities to identify diseased chickens and to use these reagents to better understand disease process.
Technical Abstract: Tumor necrosis factor-like ligand 1A (TL1A) is a type II transmembrane protein predominantly expressed by endothelial cells that promotes the expansion of activated T cells and regulatory T cells, modulates inflammation, and regulates the production of a wide variety of T cell cytokines. However, there have not been any mAbs which specifically detect chTL1A and define its biochemical and immunological properties. So in this study, two mouse monoclonal antibodies (mAbs) which specifically detect chicken TL1A (chTL1A) were developed and characterized. Both mAbs identified a 32 kDa E. coli-derived, poly-histidine-tagged fusion protein by Western blot analysis. The mAbs identified TL1A-secreting cells in the chicken thymus, cecal tonsil, and bursa of Fabricius by immunocytochemistry, and were used to measure serum TL1A levels in normal and necrotic enteritis (NE)-afflicted chickens by antigen capture ELISA. These mAbs inhibited chTL1A-induced spleen lymphocyte proliferation, nitric oxide production by chicken macrophage cells (HD11), and blocked the cytotoxic effect of chTL1A against lymphoblastoid chicken B tumor cells (LSCC-RP9). These new mAbs that detect chTL1A will be important immune reagents for basic and applied research in poultry immunology.