Two distinct affinity binding sites for IL-1 on human cell lines

J Immunol. 1989 Aug 15;143(4):1168-74.

Abstract

We used two human cell lines, NK-like YT-C3 and an EBV-containing B cell line, 3B6, as models to study the receptor(s) for IL-1. Two distinct types of saturable binding sites were found on both cell lines at 37 degrees C. Between 1 pM and 100 pM of 125I-IL-1-alpha concentration, saturable binding sites were detected on the YT-C3 cells with a K of 4 x 10(-11) M. The K found for the IL-1-alpha binding sites on 3B6 cells was 7.5 x 10(-11) M. An additional binding curve was detected above 100 pM on YT-C3 cells with a K of 7 x 10(-9) M and on 3B6 cells with a K of 5 x 10(-9) M. Scatchard plot analysis revealed 600 sites/cell with high affinity binding and 7000 sites/cell with low affinity for YT-C3 cells and 300 sites/cell with high affinity binding and 6000 sites/cell with low affinity for 3B6 cells. At 37 degrees C, the internalization of 125I-labeled IL-1 occurred via both high and low affinity IL-1R on both YT-C3 and 3B6 cells, whereas the rates of internalization for high affinity binding sites on YT-C3 cells were predominant in comparison to that of low affinity binding sites. In chemical cross-linking studies of 125I-IL-1-alpha to 3B6 and YT-C3 cells, two protein bands were immunoprecipitated with Mr around 85 to 90 kDa leading to an estimation of the Mr of the IL-1R around 68 to 72 kDa. In similar experiments, the Mr found for the IL-1R expressed on the murine T cell line EL4 was slightly higher (around 80 kDa). Whether these distinct affinity binding sites are shared by a single molecule or by various chains remains to be elucidated.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Animals
  • Binding, Competitive
  • Cross-Linking Reagents
  • Endocytosis
  • Humans
  • Interleukin-1 / metabolism*
  • Interleukin-1 / physiology
  • Iodine Radioisotopes / metabolism
  • Kinetics
  • Male
  • Mice
  • Mice, Inbred C3H
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma / metabolism
  • Receptors, Immunologic / analysis*
  • Receptors, Immunologic / physiology
  • Receptors, Interleukin-1
  • Temperature
  • Thymoma / metabolism
  • Tumor Cells, Cultured / metabolism

Substances

  • Cross-Linking Reagents
  • Interleukin-1
  • Iodine Radioisotopes
  • Receptors, Immunologic
  • Receptors, Interleukin-1