Establishment of the cells useful for murine interleukin-18 bioassay by introducing murine interleukin-18 receptor cDNA into human myelomonocytic KG-1 cells.
We genetically engineered human myelomonocytic KG-I cells by introducing cDNA of murine interleukin-18 receptor (MuIL-18R) and established human cells which were capable of responding to MuIL-18. These cells expressed larger number of MuIL-18R (> 13,000 sites/cell) than intrinsic human IL-18 receptor (HuIL-18R) (< 2,500 sites/cell). And the cells responded to MuIL-18 as well as to HuIL-18 in a dose-dependent manner, and produced large amounts of interferon-gamma (IFN-gamma). We could estimate the amount of murine IL-18 based on the amounts of IFN-gamma produced by these cells. The stoichiometry was observed up to 150 ng/ml of MuIL-18. By using these cells, a large amount of MuIL-18 (448 +/- 89.2 ng/ml) was detected in sera of Propionibacterium acnes (P. acnes)/lipopolysaccharide (LPS)-treated endotoxic mice (the same conditions in which IL-18 was first identified). These cells provide us with a useful tool for determining the bioactivity of MuIL-18.
Fujisaki Institute, Hayashibara Biochemical Laboratories Inc., Okayama, Japan.
M
M
Taniguchi
K
K
Nagaoka
S
S
Ushio
Y
Y
Nukada
T
T
Okura
T
T
Mori
H
H
Yamauchi
T
T
Ohta
H
H
Ikegami
M
M
Kurimoto
D000818:Animals; D001681:Biological Assay; D018076:DNA, Complementary / Q000235:genetics*; D019446:Endotoxemia / Q000097:blood* / Q000139:chemically induced / Q000276:immunology; D005822:Genetic Vectors; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis; D020382:Interleukin-18 / Q000032:analysis* / Q000378:metabolism; D053727:Interleukin-18 Receptor alpha Subunit; D008070:Lipopolysaccharides / Q000633:toxicity; D051379:Mice; D008810:Mice, Inbred C57BL; D009000:Monocytes / Q000378:metabolism; D011425:Propionibacterium acnes; D011485:Protein Binding; D018123:Receptors, Interleukin / Q000235:genetics / Q000378:metabolism*; D053724:Receptors, Interleukin-18; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015398:Signal Transduction; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; C508572:IL18R1 protein, human; C508573:Il18r1 protein, mouse; D020382:Interleukin-18; D053727:Interleukin-18 Receptor alpha Subunit; D008070:Lipopolysaccharides; D018123:Receptors, Interleukin; D053724:Receptors, Interleukin-18; D011993:Recombinant Fusion Proteins; D007371:Interferon-gamma