Purification and characterization of the human interleukin-18 receptor.
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.
Torigoe
K
K
Fujisaki Institute, Hayashibara Biochemical Laboratories, Inc., 675-1 Fujisaki, Okayama, 702 Japan.
Ushio
S
S
Okura
T
T
Kobayashi
S
S
Taniai
M
M
Kunikata
T
T
Murakami
T
T
Sanou
O
O
Kojima
H
H
Fujii
M
M
Ohta
T
T
Ikeda
M
M
Ikegami
H
H
Kurimoto
M
M
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D019556:COS Cells; D002462:Cell Membrane / Q000737:chemistry; D016207:Cytokines / Q000378:metabolism*; D006689:Hodgkin Disease / Q000378:metabolism; D006801:Humans; D007375:Interleukin-1 / Q000378:metabolism; D020382:Interleukin-18; D053727:Interleukin-18 Receptor alpha Subunit; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism; D018123:Receptors, Interleukin / Q000737:chemistry / Q000302:isolation & purification*; D053724:Receptors, Interleukin-18; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D016207:Cytokines; C508572:IL18R1 protein, human; C508573:Il18r1 protein, mouse; D007375:Interleukin-1; D020382:Interleukin-18; D053727:Interleukin-18 Receptor alpha Subunit; D016328:NF-kappa B; D018123:Receptors, Interleukin; D053724:Receptors, Interleukin-18