A novel recombinant fusion protein encoding a 20-amino acid residue of the third extracellular (E3) domain of CCR2 neutralizes the biological activity of CCL2

J Immunol. 2009 Jul 1;183(1):732-9. doi: 10.4049/jimmunol.0802746. Epub 2009 Jun 17.

Abstract

CCL2 is a key CC chemokine that has been implicated in a variety of inflammatory autoimmune diseases and in tumor progression and it is therefore an important target for therapeutic intervention in these diseases. Soluble receptor-based therapy is a known approach for neutralizing the in vivo functions of soluble mediators. Owing to the complexity of seven-transmembrane G protein-coupled receptors, efforts to generate neutralizing soluble chemokine receptors have so far failed. We developed a strategy that is based on the generation of short recombinant proteins encoding different segments of a G protein-coupled receptor, and tested the ability of each of them to bind and neutralize its target chemokine. We show that a fusion protein comprised of as few as 20 aa of the third extracellular (E3) domain of the CCL2 receptor, stabilized by the IgG H chain Fc domain (E3-IgG or BL-2030), selectively binds CCL2 and CCL16 and effectively neutralizes their biological activities. More importantly, E3-IgG (BL-2030) could effectively suppress the in vivo biological activity of CCL2, attenuating ongoing experimental autoimmune encephalomyelitis, as well as the development of human prostate tumor in SCID mice.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Cell Line, Tumor
  • Cell Migration Inhibition / immunology
  • Cell Proliferation
  • Chemokine CCL2 / antagonists & inhibitors*
  • Chemokine CCL2 / metabolism
  • Chemokine CCL2 / physiology*
  • Female
  • Humans
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Mice, SCID
  • Prostatic Neoplasms / immunology
  • Prostatic Neoplasms / therapy
  • Protein Binding / immunology
  • Protein Structure, Tertiary
  • Receptors, CCR2 / metabolism
  • Receptors, CCR2 / physiology*
  • Recombinant Fusion Proteins / chemical synthesis
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Fusion Proteins / physiology*

Substances

  • CCL2 protein, human
  • CCR2 protein, human
  • Chemokine CCL2
  • Receptors, CCR2
  • Recombinant Fusion Proteins