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

Lee, S. H., Lillehoj, H. S., Jeong, M. S., Del Cacho, E., Kim, J. B., Kim, H. R., Min, W., Jeoung, H. Y., An, D. J.
Poultry science 2014 v.93 no.9 pp. 2193-2198
Escherichia coli, Western blotting, amino acid sequences, binding proteins, bursa of Fabricius, chickens, ducks, geese, immunohistochemistry, interleukin-1beta, maltose, mice, molecular weight, monoclonal antibodies, nitric oxide, pigeons, recombinant fusion proteins, sequence homology, spleen, thymocytes, tonsils, turkeys
Interleukin-1β 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-1β. There have been no antibodies that specifically detect avian IL-1β, and the current study was conducted to develop mouse monoclonal antibodies (mAb) against chicken IL-1β (chIL-1β) to further define its biochemical and immunological properties. In this study, 2 mouse mAb that are specific for chIL-1β were produced and characterized. Both mAb identified a 66.0 kDa recombinant chIL-1β 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-1β protein and a 43.0 kDa maltose binding protein tag. Immunohistochemical analysis identified cells producing endogenous chIL-1β in the cecal tonsils, bursa of Fabricius, and spleen. Purified recombinant chIL-1β dose-dependently stimulated the proliferation and nitric oxide production by thymocytes, and both activities were inhibited by co-incubation with the 2 chIL-1β mAb described in this paper. These mAb will be important immune reagents for basic and applied poultry research of IL-1β in poultry.