The designer cytokine hyper-interleukin-6 is a potent activator of STAT3-dependent gene transcription in vivo and in vitro

J Biol Chem. 1999 Jan 15;274(3):1257-66. doi: 10.1074/jbc.274.3.1257.

Abstract

Interleukin-6 (IL-6) triggers pivotal pathways in vivo. The designer protein hyper-IL-6 (H-IL-6) fuses the soluble IL-6 receptor (sIL-6R) through an intermediate linker with IL-6. The intracellular pathways that are triggered by H-IL-6 are not defined yet. Therefore, we studied the molecular mechanisms leading to H-IL-6-dependent gene activation. H-IL-6 stimulates haptoglobin mRNA expression in HepG2 cells, which is transcriptionally mediated as assessed by run-off experiments. The increase in haptoglobin gene transcription correlates with higher nuclear translocation of tyrosine-phosphorylated STAT3 and its DNA binding. As H-IL-6 stimulates STAT3-dependent gene transcription, we compared the molecular mechanism between IL-6 and H-IL-6. Transfection experiments were performed with a STAT3-dependent luciferase construct. The same amount of H-IL-6 stimulated luciferase activity faster, stronger, and for a longer period of time. Dose response experiments showed that a 10-fold lower dose of H-IL-6 stimulated STAT3-dependent gene transcription comparable with the higher amount of IL-6. Cotransfection with the gp80 and/or gp130 receptor revealed that the effect of H-IL-6 on STAT3-dependent gene transcription is restricted to the gp80/gp130 receptor ratio. High amounts of gp130 increased and high amounts of gp80 decreased the effect on H-IL-6-dependent gene transcription. To investigate the in vivo effect of H-IL-6 on gene transcription in the liver, H-IL-6 and IL-6 were injected into C3H mice. H-IL-6 was at least 10-fold more effective in stimulating the DNA binding and nuclear translocation of STAT3, which enhances haptoglobin mRNA and protein expression. Thus H-IL-6 stimulates STAT3-dependent gene transcription in liver cells in vitro and in vivo at least 10-fold more effectively than IL-6. Our results provide evidence that H-IL-6 is a promising designer protein for therapeutic intervention during different pathophysiological conditions also in humans.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, CD / metabolism
  • Biological Transport
  • Cell Line
  • Cell Nucleus / metabolism
  • Cytokine Receptor gp130
  • DNA / metabolism
  • DNA-Binding Proteins / physiology*
  • Gene Expression Regulation / drug effects*
  • Haptoglobins / genetics
  • Humans
  • Interleukin-6 / metabolism*
  • Interleukin-6 / pharmacology*
  • Membrane Glycoproteins / metabolism
  • Mice
  • Mice, Inbred C3H
  • Phosphorylation
  • RNA, Messenger / metabolism
  • Receptors, Interleukin / metabolism*
  • Receptors, Interleukin-6 / metabolism
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Fusion Proteins / pharmacology
  • STAT3 Transcription Factor
  • Trans-Activators / physiology*
  • Transcription, Genetic*
  • Transcriptional Activation
  • Transfection
  • Tyrosine / metabolism

Substances

  • Antigens, CD
  • DNA-Binding Proteins
  • Haptoglobins
  • IL6ST protein, human
  • Il6st protein, mouse
  • Interleukin-6
  • Membrane Glycoproteins
  • RNA, Messenger
  • Receptors, Interleukin
  • Receptors, Interleukin-6
  • Recombinant Fusion Proteins
  • STAT3 Transcription Factor
  • STAT3 protein, human
  • Stat3 protein, mouse
  • Trans-Activators
  • interleukin 6-interleukin 6 receptor fusion protein, recombinant
  • interleukin-6 receptor alpha
  • Cytokine Receptor gp130
  • Tyrosine
  • DNA