Characterization of a Novel Interleukin-1 Receptor Antagonist from Sheep (Ovis aries)

Abstract

Interleukin-1 receptor antagonist (IL-1Ra) is an antagonist of IL-1β binding IL-1β receptors but does not induce intracellular responses or signal transduction. In this study, the full-length complementary DNA (cDNA) of the IL-1Ra gene (OaIL-1Ra) was identified from sheep (Ovis aries) using rapid amplification of cDNA ends PCR and submitted to GenBank with the accession number KC425613. The OaIL-1Ra cDNA comprised an open reading frame of 525 bp encoding a protein of 19765.8 Da, a 5'-untranslated region (UTR) of 27 bp, and a 3'-UTR of 676 bp with a poly(A) tail. Recombinant OaIL-1Ra with bioactivity was expressed in a prokaryotic expression system, and a monoclonal antibody against native OaIL-1Ra was prepared. Through Western blot analyses, the OaIL-1Ra protein was widely expressed in lung, heart, spleen, liver, kidney, muscle, intestine, lymphonodi, rumen, and white blood cells, with the highest levels in liver and spleen. The expression of OaIL-1Ra in primary cultured white blood cells of sheep were highly induced in a time-dependent manner when challenged with different bacteria. These results implied that OaIL-1Ra is associated with immune responses during bacterial infections.

Keywords: Interleukin-1 receptor antagonist; Ovis aries; bacterial infection; bioactivity; gene expression; molecular characteristics; monoclonal antibody.