Largent B L, Walton K M, Hoppe C A, Lee Y C, Schnaar R L
J Biol Chem. 1984 Feb 10;259(3):1764-9.
The ability of rabbit alveolar macrophages to specifically recognize and adhere to surfaces derivatized with carbohydrates was examined. Otherwise inert polyacrylamide gels were derivatized with aminohexylglycosides as previously described (Guarnaccia, S. P., and Schnaar, R. L. (1982) J. Biol. Chem. 257, 14288-14292). Intact viable rabbit alveolar macrophages, isolated by lung lavage, were placed in contact with surfaces derivatized with different glycosides. Only those surfaces derivatized with alpha-D-mannose residues were capable of supporting rabbit alveolar macrophage adhesion. Adhesion was rapid, obtaining maximal levels within 10 min, and occurred readily at either 0 or 37 degrees C. The carbohydrate specificity of the cell adhesion was investigated by the use of soluble carbohydrate inhibitors. The potency of various saccharides to block the adhesion correlated with that demonstrated for blocking the uptake or binding of radiolabeled soluble glycoproteins (Shepherd, V. L., Lee, Y. C., Schlesinger, P. H., and Stahl, P. D. (1981) Proc. Natl. Acad. Sci. U.S.A. 78, 1019-1022). Thus, the order of potency observed was: D-Man congruent to L-Fuc greater than D-GlcNAc congruent to D-Glc much greater than D-Gal congruent to D-GalNAc congruent to L-rhamnose. While soluble monosaccharides were capable of blocking adhesion when added in millimolar concentrations, polymannosylated neoglycoproteins were able to block adhesion in the nanomolar concentration range. Adhesion to the mannose-derivatized surfaces was a dynamic event even at 0 degrees C, since adhesion was less susceptible to monosaccharide inhibition at later incubation times. Surfaces derivatized with aminohexyl S-mannoside ligands were more effective in supporting adhesion than those derivatized with the corresponding O-mannosides. Soluble inhibitor studies suggest that this was due to a more favorable conformation of the S-glycoside for binding to the cell surface receptor. The results reported here demonstrate that the previously reported alveolar macrophage mannose/fucose receptor can mediate carbohydrate-specific cell adhesion.
对兔肺泡巨噬细胞特异性识别并黏附于碳水化合物衍生化表面的能力进行了检测。如前所述(瓜尔纳恰,S.P.,和施纳尔,R.L.(1982年)《生物化学杂志》257,14288 - 14292),用氨基己基糖苷对原本惰性的聚丙烯酰胺凝胶进行衍生化处理。通过肺灌洗分离得到的完整存活兔肺泡巨噬细胞,与用不同糖苷衍生化的表面接触。只有那些用α - D - 甘露糖残基衍生化的表面能够支持兔肺泡巨噬细胞黏附。黏附迅速,在10分钟内达到最高水平,且在0或37℃时均能轻易发生。通过使用可溶性碳水化合物抑制剂来研究细胞黏附的碳水化合物特异性。各种糖类阻断黏附的效力与阻断放射性标记可溶性糖蛋白摄取或结合所显示的效力相关(谢泼德,V.L.,李,Y.C.,施莱辛格,P.H.,和斯塔尔,P.D.(1981年)《美国国家科学院院刊》78,1019 - 1022)。因此,观察到的效力顺序为:D - 甘露糖≈L - 岩藻糖>D - N - 乙酰葡糖胺≈D - 葡萄糖>>D - 半乳糖≈D - N - 乙酰半乳糖胺≈L - 鼠李糖。虽然可溶性单糖在以毫摩尔浓度添加时能够阻断黏附,但多聚甘露糖基化新糖蛋白在纳摩尔浓度范围内就能阻断黏附。即使在0℃时,对甘露糖衍生化表面的黏附也是一个动态过程,因为在后续孵育时间,黏附对单糖抑制的敏感性降低。用氨基己基S - 甘露糖苷配体衍生化的表面在支持黏附方面比用相应O - 甘露糖苷衍生化的表面更有效。可溶性抑制剂研究表明,这是由于S - 糖苷与细胞表面受体结合的构象更有利。此处报道的结果表明,先前报道的肺泡巨噬细胞甘露糖/岩藻糖受体可介导碳水化合物特异性细胞黏附。