Oligolysine-based oligosaccharide clusters - Selective recognition and endocytosis by the mannose receptor and dendritic cell-specific intercellular adhesion molecule 3 (ICAM-3)-grabbing nonintegrin
Oligolysine-based oligosaccharide clusters - Selective recognition and endocytosis by the mannose receptor and dendritic cell-specific intercellular adhesion molecule 3 (ICAM-3)-grabbing nonintegrin
复制标题
DOI：
10.1074/jbc.m302483200
复制
发表时间：
2003-06-27
影响因子：
4.8
通讯作者：
Roche, AC
中科院分区：
文献类型：
--
作者：
Frison, N;Taylor, ME;Roche, AC
树突状细胞是一种高效的抗原提呈细胞，表达多种膜凝集素，包括甘露糖受体和DC-SIGN(树突状细胞特异性ICAM-3-抓取非整合素)。为了确定这些树突状细胞受体的高度特异性配体，将寡糖转化为糖合成子(Os-1)，并用于制备基于寡聚赖氨酸的糖簇，Os-[Lys(Os)](N)-Ala-Cys-NH2。合成了含有2~6个二甘露糖苷的簇合物以及含有4个或5个五糖(Lewis(A)或Lewis(X))或六糖(Lewis(B))的簇合物。附加的半胱氨酸残基的硫醇基团允许通过荧光探针容易地标记或方便地用抗原替代。表面等离子共振用于确定不同糖簇对纯化的甘露糖受体和DC-SIGN的亲和力，而流式细胞术和共聚焦显微镜分析可以评估细胞对荧光素标记的糖簇的摄取。甘露糖受体可识别二烯醇苷簇，其亲和力常数接近10(6)·m o l(-1)，但对DC-SIGN的亲和力很低(小于104 m e·m o l(-1))。相反，Lewis簇对DC-SIGN的亲和力高于对甘露糖受体的亲和力。人树突状细胞和表达甘露糖受体的大鼠成纤维细胞都能有效地摄取二芒果苷簇，但表达DC-SIGN的HeLa细胞或大鼠成纤维细胞不能有效地摄取；表达DC-SIGN的细胞可以摄取Lewis簇。结果表明，含有甘露糖苷簇的配体可以特异性地靶向甘露糖受体，而含有Lewis簇的配体将靶向DC-SIGN。
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 (Os-1) 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 (Lewis(a) or Lewis(x)) or hexasaccharides (Lewis(b)) 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 10(6) liter.mol(-1) but have a very low affinity for DC-SIGN (less than 104 liter.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.