Development and characterization of mouse monoclonal antibodies reactive with chicken TL1A

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 Escherichia 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.

Keywords: Chickens; Hybridoma; Lymphocyte; Macrophage; TL1A; TNFSF15.