Cloning, expression analyses, and chromosomal location of porcine interleukin-18 receptor alpha chain (IL-18Ralpha)

J Interferon Cytokine Res. 2002 Sep;22(9):995-1002. doi: 10.1089/10799900260286704.

Abstract

We cloned and sequenced a cDNA that contains the coding sequence of the porcine interleukin-18 receptor alpha chain (PoIL-18Ralpha). Based on the conserved nucleotide sequences between human (HuIL-18Ralpha) and murine IL-18Ralpha (MuIL-18Ralpha), we performed reverse transcription-polymerase chain reaction (RT-PCR) with total RNA prepared from porcine peripheral blood lymphocytes (PBLs) stimulated with PoIL-12 to clone the cDNA of PoIL-18Ralpha. The open reading frame (ORF) of the PoIL-18Ralpha cDNA is 1620 base pairs (bp) in length and encodes 539 amino acids. The predicted amino acid sequence showed 68.2% and 50.2% identity to the human and murine amino acid sequences, respectively. Stimulation with concanavalin A (ConA) and IL-12, but not with IL-4, was shown to upregulate the expression of IL-18Ralpha mRNA in pig PBLs by RT-PCR analysis. Flow cytometric analysis also demonstrated that IL-18Ralpha was constitutively expressed on PoPBLs, and this expression was augmented by ConA stimulation. Furthermore, the PoIL-18Ralpha gene was mapped by fluorescence in situ hybridization (FISH) to porcine chromosome 3 (3q13-q14), near the location at which the IL-1beta gene had already been mapped. The present results will be helpful for understanding PoIL-18 and interferon gamma (IFN-gamma)-mediated T helper 1 (Th1) cell development.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Chromosome Mapping / veterinary*
  • Cloning, Molecular
  • Concanavalin A / pharmacology
  • DNA, Complementary / genetics
  • DNA, Complementary / isolation & purification
  • Gene Expression Regulation / drug effects*
  • Humans
  • In Situ Hybridization, Fluorescence
  • Interleukin-12 / pharmacology
  • Interleukin-18 Receptor alpha Subunit
  • Interleukin-4 / pharmacology
  • Lymphocytes / drug effects
  • Lymphocytes / metabolism
  • Mice
  • Molecular Sequence Data
  • Open Reading Frames
  • Protein Subunits / genetics
  • RNA, Messenger / biosynthesis
  • Radiation Hybrid Mapping
  • Receptors, Interleukin / biosynthesis
  • Receptors, Interleukin / genetics*
  • Receptors, Interleukin-18
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Species Specificity
  • Swine / genetics*

Substances

  • DNA, Complementary
  • IL18R1 protein, human
  • Il18r1 protein, mouse
  • Interleukin-18 Receptor alpha Subunit
  • Protein Subunits
  • RNA, Messenger
  • Receptors, Interleukin
  • Receptors, Interleukin-18
  • Concanavalin A
  • Interleukin-12
  • Interleukin-4