Molecular cloning and functional characterization of avian interleukin-19

Sungwon Kim, Kate B Miska, Audrey P McElroy, Mark C Jenkins, Ray H Fetterer, Chasity M Cox, Lindsay H Stuard, Rami A Dalloul

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish
Pages (from-to)476-84
Number of pages9
JournalMolecular Immunology
Volume47
Issue number2-3
DOIs
Publication statusPublished - Dec 2009

Keywords

  • Amino Acid Sequence
  • Animals
  • Apoptosis
  • Chickens
  • Cloning, Molecular
  • Gene Expression Regulation
  • Inflammation Mediators
  • Interleukins
  • Molecular Sequence Data
  • Monocytes
  • Nitric Oxide
  • Phylogeny
  • Recombinant Proteins
  • Sequence Analysis, DNA
  • Sequence Homology, Amino Acid
  • Th2 Cells

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