Oligolysine-based oligosaccharide clusters: selective recognition and endocytosis by the mannose receptor and dendritic cell-specific intercellular adhesion molecule 3 (ICAM-3)-grabbing nonintegrin.
Dendritic cells are potent antigen-presenting cells that express several membrane lectins, including the mannose receptor and DC-SIGN (dendritic cell-specific ICAM-3-grabbing nonintegrin). To identify highly specific ligands for these dendritic cell receptors, oligosaccharides were converted into glycosynthons (Os1) and were used to prepare oligolysine-based glycoclusters, Os-[Lys(Os)]n-Ala-Cys-NH2. Clusters containing two to six dimannosides as well as clusters containing four or five pentasaccharides (Lewisa or Lewisx) or hexasaccharides (Lewisb) were synthesized. The thiol group of the appended cysteine residue allows easy tagging by a fluorescent probe or convenient substitution with an antigen. Surface plasmon resonance was used to determine the affinity of the different glycoclusters for purified mannose receptor and DC-SIGN, whereas flow cytometry and confocal microscopy analysis allowed assessment of cell uptake of fluoresceinyl-labeled glycoclusters. Dimannoside clusters are recognized by the mannose receptor with an affinity constant close to 106 liter.mol-1 but have a very low affinity for DC-SIGN (less than 104 liter x mol-1). Conversely, Lewis clusters have a higher affinity toward DC-SIGN than toward the mannose receptor. Dimannoside clusters are efficiently taken up by human dendritic cells as well as by rat fibroblasts expressing the mannose receptor but not by HeLa cells or rat fibroblasts expressing DC-SIGN; DC-SIGN-expressing cells take up Lewis clusters. The results suggest that ligands containing dimannoside clusters can be used specifically to target the mannose receptor, whereas ligands containing Lewis clusters will be targeted to DC-SIGN.
Frison
Natacha
N
Glycobiologie-Vectorologie et Trafic Intracellulaire, Centre de Biophysique Moléculaire, CNRS, Rue Charles-Sadron, 45071 Orléans cedex 02, France.
Taylor
Maureen E
ME
Soilleux
Elizabeth
E
Bousser
Marie-Thérèse
MT
Mayer
Roger
R
Monsigny
Michel
M
Drickamer
Kurt
K
Roche
Annie-Claude
AC
D019264:Adoptive Transfer / Q000379:methods; D015815:Cell Adhesion Molecules / Q000378:metabolism; D003713:Dendritic Cells / Q000737:chemistry / Q000276:immunology; D004705:Endocytosis; D005434:Flow Cytometry; D005643:Fucose; D006801:Humans; D037102:Lectins / Q000378:metabolism; D037181:Lectins, C-Type / Q000378:metabolism; D008024:Ligands; D008239:Lysine; D008358:Mannose; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins / Q000378:metabolism; D009844:Oligosaccharides / Q000138:chemical synthesis* / Q000493:pharmacokinetics* / Q000627:therapeutic use; D011956:Receptors, Cell Surface / Q000378:metabolism; D020349:Surface Plasmon Resonance; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
D015815:Cell Adhesion Molecules; D005643:Fucose; D037102:Lectins; D037181:Lectins, C-Type; D008024:Ligands; D008239:Lysine; D008358:Mannose; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins; D009844:Oligosaccharides; D011956:Receptors, Cell Surface; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin