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

Poult Sci. 2014 Sep;93(9):2193-8. doi: 10.3382/ps.2014-03947. Epub 2014 Jul 18.

Abstract

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.

Keywords: chicken; interleukin 1β; monoclonal antibody; proliferation; thymus.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology*
  • Avian Proteins / genetics
  • Avian Proteins / immunology*
  • Blotting, Western
  • Chickens / genetics
  • Chickens / immunology*
  • Enzyme-Linked Immunosorbent Assay / veterinary
  • Interleukin-1beta / genetics
  • Interleukin-1beta / immunology*
  • Lymphoid Tissue
  • Mice / immunology
  • Mice, Inbred BALB C
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology

Substances

  • Antibodies, Monoclonal
  • Avian Proteins
  • Interleukin-1beta
  • Recombinant Fusion Proteins