Cloning of the chicken interleukin-13 receptor alpha 2 gene and production of a specific monoclonal antibody

Dev Comp Immunol. 2007;31(4):394-406. doi: 10.1016/j.dci.2006.07.002. Epub 2006 Aug 7.

Abstract

Mammalian interleukin-13 (IL-13) is an important regulatory T2 cytokine secreted by activated T lymphocytes. The IL-13 receptor (IL-13R) has two different chains, IL-13Ralpha1 and IL-13Ralpha2. Although the chicken IL-13 gene is well characterized, little is known about IL-13Rs. We cloned a cDNA encoding the 380 amino acid pro-peptide of chicken IL-13Ralpha2 (chIL-13Ralpha2) and developed a monoclonal antibody (mAb), HU13-1, against it. The chIL-13Ralpha2 amino acid sequence showed 37-39% sequence identity with mammalian homologs. High levels of chIL-13Ralpha2 mRNA were expressed in liver, testis, ovary, brain, and lipopolysaccharide (LPS)-stimulated IN24 cells. HU13-1 specifically recognized recombinant chIL-13Ralpha2 in ELISAs, and western blots identified a 45-kDa glycoprotein or a 41-kDa non-glycosylated protein in LPS-stimulated IN24 cell lysates. LPS induced a gradual increase in HU13-1-positive IN24 cells over 20 h. These results indicate that mAb HU13-1 recognizes native chIL-13Ralpha2 and will be valuable for further studies of chIL-13Rs.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal / biosynthesis*
  • Antibody Specificity*
  • Base Sequence
  • Cell Line, Tumor
  • Chickens / genetics
  • Chickens / immunology*
  • Cloning, Molecular*
  • Female
  • Humans
  • Interleukin-13 Receptor alpha2 Subunit / genetics*
  • Interleukin-13 Receptor alpha2 Subunit / immunology*
  • Mice
  • Mice, Inbred BALB C
  • Molecular Sequence Data
  • Organ Specificity / genetics
  • Organ Specificity / immunology

Substances

  • Antibodies, Monoclonal
  • Interleukin-13 Receptor alpha2 Subunit

Associated data

  • GENBANK/AB243738