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

J Immunol Methods. 1998 Aug 1;217(1-2):97-102. doi: 10.1016/s0022-1759(98)00098-2.

Abstract

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.

MeSH terms

  • Animals
  • Biological Assay*
  • DNA, Complementary / genetics*
  • Endotoxemia / blood*
  • Endotoxemia / chemically induced
  • Endotoxemia / immunology
  • Genetic Vectors
  • Humans
  • Interferon-gamma / biosynthesis
  • Interleukin-18 / analysis*
  • Interleukin-18 / metabolism
  • Interleukin-18 Receptor alpha Subunit
  • Lipopolysaccharides / toxicity
  • Mice
  • Mice, Inbred C57BL
  • Monocytes / metabolism
  • Propionibacterium acnes
  • Protein Binding
  • Receptors, Interleukin / genetics
  • Receptors, Interleukin / metabolism*
  • Receptors, Interleukin-18
  • Recombinant Fusion Proteins / metabolism
  • Signal Transduction
  • Tumor Cells, Cultured

Substances

  • DNA, Complementary
  • IL18R1 protein, human
  • Il18r1 protein, mouse
  • Interleukin-18
  • Interleukin-18 Receptor alpha Subunit
  • Lipopolysaccharides
  • Receptors, Interleukin
  • Receptors, Interleukin-18
  • Recombinant Fusion Proteins
  • Interferon-gamma