An assay for functional dendritic cell-specific ICAM-3-grabbing nonintegrin (DC-SIGN) inhibitors of human dendritic cell adhesion

Anal Biochem. 2010 Nov 15;406(2):222-9. doi: 10.1016/j.ab.2010.07.018. Epub 2010 Jul 25.

Abstract

We report a new dendritic cell adhesion assay, using either immature or mature dendritic cells, for identifying functional dendritic cell-specific ICAM-3-grabbing nonintegrin (DC-SIGN) inhibitors. Because immature dendritic cells are responsible for pathogen binding and invasion, this in vitro assay provides an important link between in vitro results and pathogen-based in vivo assays. Furthermore, this assay does not require laborious expression, refolding, and purification of DC-SIGN carbohydrate recognition domain or extracellular domain as receptor-based assays. The assay power evaluated with Z and Z' parameters enables screening of compound libraries and determination of IC(50) values in the first stage of DC-SIGN inhibitor development.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibody Specificity / immunology
  • Cell Adhesion
  • Cell Adhesion Molecules / antagonists & inhibitors*
  • Cell Adhesion Molecules / metabolism
  • Cell Differentiation
  • Colorimetry
  • Dendritic Cells / cytology*
  • Dendritic Cells / metabolism*
  • Flow Cytometry
  • Fluoresceins / metabolism
  • Fluorescence
  • Humans
  • Immunoassay / methods*
  • Inhibitory Concentration 50
  • Lectins, C-Type / antagonists & inhibitors*
  • Lectins, C-Type / metabolism
  • Ligands
  • Receptors, Cell Surface / antagonists & inhibitors*
  • Receptors, Cell Surface / metabolism
  • Reproducibility of Results
  • Succinimides / metabolism
  • Time Factors

Substances

  • 5-(6)-carboxyfluorescein diacetate succinimidyl ester
  • Cell Adhesion Molecules
  • DC-specific ICAM-3 grabbing nonintegrin
  • Fluoresceins
  • Lectins, C-Type
  • Ligands
  • Receptors, Cell Surface
  • Succinimides