Molecular cloning and characterization of a ruminant interleukin-6 cDNA

By hybridization with a human interleukin-6 (IL-6) cDNA fragment a corresponding ruminant (ovine) cDNA was isolated from a lipopolysaccharide (LPS)-stimulated alveolar macrophage library. The nucleotide sequence of the cDNA and the predicted amino acid sequence of the protein showed significant homology to the human and murine molecules. Ovine IL-6 cDNA encodes a polypeptide of 208 amino acids that, based on analysis of human IL-6, is processed to a protein of 180 amino acids. Northern blot analysis and the 7TD1 bioassay were used to analyse regulatory aspects of IL-6 production by primary ovine fibroblasts. Both LPS and recombinant ovine IL-1 alpha were shown to induce IL-6 mRNA with peak levels occurring at 1 h post-stimulation and declining thereafter. When fibroblasts were pretreated with cyclohexamide prior to stimulation the level of induction by LPS and IL-1 alpha increased dramatically and peak levels were observed at 5 h post-stimulation. The level of secreted IL-6 increased rapidly over the first 24 h and continued to increase over the next 48 h.