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

摘要

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-13R alpha 1 and IL-13R alpha 2. Although the chicken IL-13 gene is well characterized, little is known about IL-13R.s. We cloned a cDNA encoding the 380 amino acid pro-peptide of chicken IL-13R alpha 2 (chIL-13R alpha 2) and developed a monoclonal antibody (mAb), HU13-1, against it. The chIL-13R alpha 2 amino acid sequence showed 37-39% sequence identity with mammalian homologs. High levels of chIL-13 R alpha 2 m RNA were expressed in liver, testis, ovary, brain, and lipopolysaccharide (LPS)-stimulated 1N24 cells. HU13-1 specifically recognized recombinant chIL-13R alpha 2 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-I-positive IN24 cells over 20 h. These results indicate that mAb HU13-1 recognizes native chIL-13R alpha 2 and will be valuable for further studies of chIL-13Rs. (c) 2006 Elsevier Ltd. All rights reserved.