Development of improved soluble inhibitors of FasL and CD40L based on oligomerized receptors

J Immunol Methods. 2000 Apr 3;237(1-2):159-73. doi: 10.1016/s0022-1759(99)00239-2.

Abstract

TNF receptor family members fused to the constant domain of immunoglobulin G have been widely used as immunoadhesins in basic in vitro and in vivo research and in some clinical applications. In this study, we assemble soluble, high avidity chimeric receptors on a pentameric scaffold derived from the coiled-coil domain of cartilage oligomeric matrix protein (COMP). The affinity of Fas and CD40 (but not TNFR-1 and TRAIL-R2) to their ligands is increased by fusion to COMP, when compared to the respective Fc chimeras. In functional assays, Fas:COMP was at least 20-fold more active than Fas:Fc at inhibiting the action of sFasL, and CD40:COMP could block CD40L-mediated proliferation of B cells, whereas CD40:Fc could not. In conclusion, members of the TNF receptor family can display high specificity and excellent avidity for their ligands if they are adequately multimerized.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • B-Lymphocytes / cytology
  • B-Lymphocytes / drug effects
  • B-Lymphocytes / immunology
  • CD40 Antigens / metabolism
  • CD40 Ligand
  • Cartilage Oligomeric Matrix Protein
  • Extracellular Matrix Proteins / genetics
  • Extracellular Matrix Proteins / metabolism
  • Extracellular Matrix Proteins / pharmacology
  • Fas Ligand Protein
  • Glycoproteins / genetics
  • Glycoproteins / metabolism
  • Glycoproteins / pharmacology
  • Humans
  • Jurkat Cells
  • Ligands
  • Lymphocyte Activation / drug effects
  • Matrilin Proteins
  • Membrane Glycoproteins / antagonists & inhibitors*
  • Membrane Glycoproteins / metabolism
  • Mice
  • Mice, Knockout
  • Receptors, Fc / genetics
  • Receptors, Fc / metabolism
  • Receptors, Tumor Necrosis Factor / genetics
  • Receptors, Tumor Necrosis Factor / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Fusion Proteins / pharmacology
  • Solubility
  • Tumor Cells, Cultured
  • fas Receptor / metabolism

Substances

  • CD40 Antigens
  • Cartilage Oligomeric Matrix Protein
  • Extracellular Matrix Proteins
  • FASLG protein, human
  • Fas Ligand Protein
  • Fasl protein, mouse
  • Glycoproteins
  • Ligands
  • Matn1 protein, mouse
  • Matrilin Proteins
  • Membrane Glycoproteins
  • Receptors, Fc
  • Receptors, Tumor Necrosis Factor
  • Recombinant Fusion Proteins
  • TSP5 protein, human
  • fas Receptor
  • CD40 Ligand