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.
Copyright 2010 Elsevier Inc. All rights reserved.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Antibody Specificity / immunology
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Cell Adhesion
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Cell Adhesion Molecules / antagonists & inhibitors*
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Cell Adhesion Molecules / metabolism
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Cell Differentiation
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Colorimetry
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Dendritic Cells / cytology*
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Dendritic Cells / metabolism*
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Flow Cytometry
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Fluoresceins / metabolism
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Fluorescence
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Humans
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Immunoassay / methods*
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Inhibitory Concentration 50
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Lectins, C-Type / antagonists & inhibitors*
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Lectins, C-Type / metabolism
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Ligands
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Receptors, Cell Surface / antagonists & inhibitors*
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Receptors, Cell Surface / metabolism
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Reproducibility of Results
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Succinimides / metabolism
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Time Factors
Substances
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5-(6)-carboxyfluorescein diacetate succinimidyl ester
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Cell Adhesion Molecules
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DC-specific ICAM-3 grabbing nonintegrin
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Fluoresceins
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Lectins, C-Type
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Ligands
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Receptors, Cell Surface
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Succinimides