Canine interleukin-13: molecular cloning of full-length cDNA and expression of biologically active recombinant protein

• Published in: Journal of interferon & cytokine research Vol. 20; no. 9; pp. 779 - 785
• Main Authors: Yang, S, Boroughs, K L, McDermott, M J
• Format: Journal Article
• Language: English
• Published: United States Mary Ann Liebert, Inc 01.09.2000
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Cells, Cultured; Cloning, Molecular; DNA, Complementary - analysis; DNA, Complementary - genetics; Dogs; Escherichia coli; Humans; Interleukin-13 - biosynthesis; Interleukin-13 - genetics; Interleukin-13 - immunology; Interleukin-13 - pharmacology; Leukocytes, Mononuclear - physiology; Lymph Nodes - physiology; Molecular Sequence Data; Rats; Recombinant Proteins - biosynthesis; Recombinant Proteins - immunology; Recombinant Proteins - pharmacology; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - analysis; Sequence Homology, Amino Acid; Transfection
• ISSN: 1079-9907; 1557-7465
• DOI: 10.1089/10799900050151049

Interleukin-13 (IL-13) regulates immune responses mediated by type 2 T helper lymphocytes (Th2) in the human and mouse. To study the function of this cytokine in the dog, we have isolated a cDNA that encodes the full-length canine IL-13 (CaIL-13) precursor polypeptide of 131 amino acids. CaIL-13 shares significant homology with the IL-13 amino acid sequences of cattle (54.1%), mouse (39.6%), and rat (36.6%) but shares the highest identity with human IL-13 (HuIL-13) (61.8%). The predicted CaIL-13 mature polypeptide of 111 residues was expressed in bacteria, and recombinant CaIL-13 (rCaIL-13) was isolated from inclusion bodies and refolded. rCaIL-13 stimulated the proliferation of TF-1 cells, which are derived from human erythroleukemia cells and respond to IL-13 as well as to a number of other human and murine cytokines. CaIL-13 mRNA was readily detectable by reverse transcriptase-polymerase chain reaction (RT-PCR) in cells from lymph nodes and peripheral blood. The gene sequence and biologically active recombinant protein for CaIL-13 will be useful reagents to determine the role of IL-13 in the regulation of canine immune responses.