Molecular cloning and functional characterization of avian interleukin-19.
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The present study describes the cloning and functional characterization of avian interleukin (IL)-19, a cytokine that, in mammals, alters the balance of Th1 and Th2 cells in favor of the Th2 phenotype. The full-length avian IL-19 gene, located on chromosome 26, was amplified from LPS-stimulated chicken monocytes, and cloned into both prokaryotic (pET28a) and eukaryotic (pcDNA3.1) expression vectors. The confirmed avian IL-19 amino acid sequence has 66.5% homology with human and murine IL-19, with a predicted protein sequence of 176 amino acids. Analysis of avian IL-19 amino acid sequence showed six conserved, structurally relevant, cysteine residues as found in mammals, but only one N-glycosylation residue. The recombinant IL-19 (rChIL-19) expressed in the prokaryotic system was purified by Ni(+)-resin column followed by endotoxin removal. Using purified avian rChIL-19, expression of Th2 cytokines was measured in splenocytes using quantitative real-time PCR (qRT-PCR). In the presence of rChIL-19, expression levels of IL-4 and IL-13, as well as IL-10, were significantly increased after 6- and 12 h treatments. This was confirmed by treating splenocytes with supernatants from IL-19 transfected cells. Also, avian monocytes incubated with rChIL-19 displayed increased expression of IL-1beta, IL-6, and IL-19. This study represents the first report for the cloning, expression, and functional characterization of avian IL-19. Taken together, avian IL-19 function seems to be conserved and similar to that of mammals and may play an important role in responses to intracellular poultry pathogens like bacteria and protozoa.
