Establishment of the cells useful for murine interleukin-18 bioassay by introducing murine interleukin-18 receptor cDNA into human myelomonocytic KG-1 cells

• Published in: Journal of immunological methods Vol. 217; no. 1; pp. 97 - 102
• Main Authors: Taniguchi, Mutsuko, Nagaoka, Katsue, Ushio, Shimpei, Nukada, Yoshiyuki, Okura, Takanori, Mori, Tsutomu, Yamauchi, Hiroshi, Ohta, Tsunetaka, Ikegami, Hakuo, Kurimoto, Masashi
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.08.1998; Elsevier
• Subjects: Animal cells; Animals; Bioassay; Biological and medical sciences; Biological Assay; Cell cultures. Hybridization. Fusion; Cytokine; DNA, Complementary - genetics; Endotoxemia - blood; Endotoxemia - chemically induced; Endotoxemia - immunology; Fundamental and applied biological sciences. Psychology; Genetic Vectors; Human cells; Humans; Interferon-gamma - biosynthesis; Interleukin-18 - analysis; Interleukin-18 - metabolism; Interleukin-18 Receptor alpha Subunit; Lipopolysaccharides - toxicity; Mice; Mice, Inbred C57BL; Molecular and cellular biology; Monocytes - metabolism; Murine IL-18; Propionibacterium acnes; Protein Binding; Receptors, Interleukin - genetics; Receptors, Interleukin - metabolism; Receptors, Interleukin-18; Recombinant DNA; Recombinant Fusion Proteins - metabolism; Signal Transduction; Tumor Cells, Cultured; IL-18R, interleukin-18 receptor; Bioassay; Murine IL-18; cDNA, complementary DNA; Cytokine; IFNγ, interferon-γ; Recombinant DNA; IL-18, interleukin-18; Human cells; Human; Cell line; Transfection; Established cell line; Bone marrow; Biosynthesis; Gamma interferon; Biological receptor; Genetic transfer
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/S0022-1759(98)00098-2

We genetically engineered human myelomonocytic KG-1 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-γ (IFN-γ). We could estimate the amount of murine IL-18 based on the amounts of IFN-γ 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.