An assay for functional dendritic cell-specific ICAM-3-grabbing nonintegrin (DC-SIGN) inhibitors of human dendritic cell adhesion.
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.
Obermajer
Natasa
N
Department of Biotechnology, Jozef Stefan Institute, SI-1000 Ljubljana, Slovenia.
Svajger
Urban
U
Jeras
Matjaz
M
Sattin
Sara
S
Bernardi
Anna
A
Anderluh
Marko
M
D000918:Antibody Specificity / Q000276:immunology; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000037:antagonists & inhibitors* / Q000378:metabolism; D002454:Cell Differentiation; D003124:Colorimetry; D003713:Dendritic Cells / Q000166:cytology* / Q000378:metabolism*; D005434:Flow Cytometry; D005452:Fluoresceins / Q000378:metabolism; D005453:Fluorescence; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D020128:Inhibitory Concentration 50; D037181:Lectins, C-Type / Q000037:antagonists & inhibitors* / Q000378:metabolism; D008024:Ligands; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015203:Reproducibility of Results; D013388:Succinimides / Q000378:metabolism; D013997:Time Factors; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
D015815:Cell Adhesion Molecules; D005452:Fluoresceins; D037181:Lectins, C-Type; D008024:Ligands; D011956:Receptors, Cell Surface; D013388:Succinimides; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin; C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester