Purification and characterization of the human interleukin-18 receptor

J Biol Chem. 1997 Oct 10;272(41):25737-42. doi: 10.1074/jbc.272.41.25737.

Abstract

Interleukin (IL)-18 was identified as a molecule that induces IFN-gamma production and enhances NK cell cytotoxicity. In this paper, we report upon the purification and characterization of human IL-18 receptor (hIL-18R). We selected the Hodgkin's disease cell line, L428, as the most strongly hIL-18R-expressing cell line based on the results of binding assays. This binding was inhibited by IL-18 but not by IL-1beta. The dissociation constant (Kd) of 125I-IL-18 binding to L428 cells was about 18.5 nM, with 18,000 binding sites/cell. After immunizing mice with L428 cells and cloning, a single monoclonal antibody (mAb) against hIL-18R was obtained (mAb 117-10C). Sequentially, hIL-18R was purified from 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid (CHAPS)-extracted L428 cells by wheat germ lectin-Sepharose 4B chromatography and mAb 117-10C-Sepharose chromatography. The internal amino acid sequences of hIL-18R all matched those of human IL-1 receptor-related protein (IL-1Rrp), the ligand of which was unknown to date. When expressed in COS-1 cells, the cDNA of IL-1Rrp conferred IL-18 binding properties on the cells and the capacity for signal transduction. From these results, we conclude that a functional IL-18 receptor component is IL-1Rrp.

Publication types

  • Comparative Study

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal
  • COS Cells
  • Cell Membrane / chemistry
  • Cytokines / metabolism*
  • Hodgkin Disease / metabolism
  • Humans
  • Interleukin-1 / metabolism
  • Interleukin-18
  • Interleukin-18 Receptor alpha Subunit
  • Kinetics
  • Mice
  • Mice, Inbred BALB C
  • Molecular Sequence Data
  • NF-kappa B / metabolism
  • Receptors, Interleukin / chemistry
  • Receptors, Interleukin / isolation & purification*
  • Receptors, Interleukin-18
  • Tumor Cells, Cultured

Substances

  • Antibodies, Monoclonal
  • Cytokines
  • IL18R1 protein, human
  • Il18r1 protein, mouse
  • Interleukin-1
  • Interleukin-18
  • Interleukin-18 Receptor alpha Subunit
  • NF-kappa B
  • Receptors, Interleukin
  • Receptors, Interleukin-18