Identification and characterization of the rat intestinal epithelial cell (IEC-18) interleukin-1 receptor

Am J Physiol. 1994 May;266(5 Pt 1):C1198-203. doi: 10.1152/ajpcell.1994.266.5.C1198.

Abstract

Interleukin-1 (IL-1), an important mediator of inflammation, may act in chronic inflammatory disorders of the intestine such as idiopathic inflammatory bowel diseases. Although the IL-1 receptor (IL-1R) has been studied extensively in many cell lines, including T cells, B cells, and fibroblasts, it has not been demonstrated on intestinal epithelial cells. IL-1 affects intestinal epithelial cell proliferation in vitro, suggesting the presence of IL-1Rs on intestinal epithelium. This paper demonstrates and further characterizes an IL-1R in a rat intestinal epithelial cell. Cells from rat intestinal epithelial cell line IEC-18 were grown to confluence in six-well plates, and association studies with 125I-labeled IL-1 beta to determine specific binding and equilibrium conditions were performed. A competitive-inhibition curve verified the IL-1 concentration required to saturate the IL-1R, and a Scatchard plot revealed a dissociation constant (Kd) of 1.2 x 10(-10) M, with 1,000 receptors per epithelial cell. Binding studies using increasing concentrations of 125I IL-1 beta alone confirmed the receptor density. Cross-linking 125I-IL-1 beta to the IEC-18 IL-1R demonstrated an IL-1R of approximately 80.5 kDa. cDNA transcribed from IEC-18 mRNA was used as a template for amplification of a segment of the IL-1R using complementary oligonucleotides. The resulting sequence of the IL-1R demonstrated a high degree of homology with both the human and mouse type I IL-1R.(ABSTRACT TRUNCATED AT 250 WORDS)

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • Binding, Competitive
  • Cell Line
  • Conserved Sequence
  • DNA Primers
  • DNA, Complementary / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Epithelium / immunology
  • Epithelium / metabolism
  • Fibroblasts / immunology
  • Fibroblasts / metabolism
  • Humans
  • Interleukin-8 / metabolism*
  • Intestines
  • Kinetics
  • Mice
  • Molecular Sequence Data
  • Molecular Weight
  • Polymerase Chain Reaction / methods
  • Rats
  • Receptors, Interleukin / genetics
  • Receptors, Interleukin / isolation & purification
  • Receptors, Interleukin / metabolism*
  • Receptors, Interleukin-8A
  • Sequence Homology, Nucleic Acid
  • T-Lymphocytes / immunology
  • T-Lymphocytes / metabolism
  • Transcription, Genetic

Substances

  • DNA Primers
  • DNA, Complementary
  • Interleukin-8
  • Receptors, Interleukin
  • Receptors, Interleukin-8A

Associated data

  • GENBANK/S70238