Development and characterization of mouse monoclonal antibodies reactive with chicken IL-1 beta

Abstract

Interleukin-1 beta proteins from chicken, duck, goose, turkey, and pigeon share 77 to 99% amino acid sequence similarity among themselves, and only 31 to 35% sequence similarity is shared between avian and mammalian IL-1 beta. There have been no antibodies that specifically detect avian IL-1 beta, and the current study was conducted to develop mouse monoclonal antibodies (mAb) against chicken IL-1 beta (chIL-1 beta) to further define its biochemical and immunological properties. In this study, 2 mouse mAb that are specific for chIL-1 beta were produced and characterized. Both mAb identified a 66.0 kDa recombinant chIL-1 beta protein expressed in Escherichia coli by Western blot analysis that corresponded to the expected molecular weight of a recombinant fusion protein containing the full-length 23.0 kDa chIL-1 beta protein and a 43.0 kDa maltose binding protein tag. Immunohistochemical analysis identified cells producing endogenous chIL-1 beta in the cecal tonsils, bursa of Fabricius, and spleen. Purified recombinant chIL-1 beta dose-dependently stimulated the proliferation and nitric oxide production by thymocytes, and both activities were inhibited by co-incubation with the 2 chIL-1 beta mAb described in this paper. These mAb will be important immune reagents for basic and applied poultry research of IL-1 beta in poultry.