Characterization of receptors for human tumour necrosis factor and their regulation by gamma-interferon

Nature. 1985 Dec 19-1986 Jan 1;318(6047):665-7. doi: 10.1038/318665a0.

Abstract

Tumour necrosis factors, TNF-alpha and TNF-beta (previously called lymphotoxin), are the products of activated monocytes and lymphocytes, respectively, and both have recently been purified, sequenced and cloned by recombinant DNA methods, revealing 35% identity and 50% homology in the amino-acid sequence. Both proteins have been found to be specifically toxic to many tumour cells. Furthermore, it has been reported that various interferons are synergistic with TNF for anti-tumour effects in vitro, while activities attributed to the two proteins have also been shown to necrotize various tumours in vivo. We have now prepared 125I-labelled highly purified recombinant human TNF-alpha to study in detail its binding to the human cervical carcinoma cell line ME-180. Our results indicate that there is a single class of specific high-affinity receptors for TNF on this cell line which has a Kd of about 0.2 nM and an average of 2,000 receptor sites per cell. The binding of labelled TNF-alpha to these cells can be inhibited by both TNF-alpha and TNF-beta but not by gamma-interferon (IFN-gamma). However, preincubation of cells with IFN-gamma increases the total number of TNF receptors two to threefold without any significant change in the affinity constant. This is the first report that TNF-alpha and -beta share a common receptor and that the receptors can be up-regulated by interferon. Our results may explain previous observations regarding similar biological activities observed for these two cytotoxic proteins and also their synergistic action with interferons.

MeSH terms

  • Cell Line
  • Female
  • Glycoproteins / metabolism*
  • Humans
  • Interferon-gamma / pharmacology*
  • Iodine / metabolism
  • Molecular Weight
  • Receptors, Cell Surface / metabolism*
  • Receptors, Tumor Necrosis Factor
  • Recombinant Proteins / metabolism
  • Substrate Specificity
  • Time Factors
  • Tumor Necrosis Factor-alpha
  • Uterine Cervical Neoplasms / analysis

Substances

  • Glycoproteins
  • Receptors, Cell Surface
  • Receptors, Tumor Necrosis Factor
  • Recombinant Proteins
  • Tumor Necrosis Factor-alpha
  • Interferon-gamma
  • Iodine