Uemura K, Macher B A, DeGregorio M, Scudder P, Buehler J, Knapp W, Feizi T
Biochim Biophys Acta. 1985 Jul 30;846(1):26-36. doi: 10.1016/0167-4889(85)90106-5.
Six monoclonal antibodies with known specificities for the carbohydrate antigens i, X or Y, and seven anti-myeloid antibodies (determinants unknown) selected for their differing reaction patterns with human leucocytes were tested in chromatogram binding assays for reactions with myeloid cell glycolipids derived from normal human granulocytes and chronic myelogenous leukemia cells. Antigenicities were found exclusively on minor glycolipids which were barely or not at all detectable with orcinol-sulphuric acid stain. Among these, a neutral glycosphingolipid bound the anti-i antibody Den and chromatographed as the ceramide octasaccharide, Gal beta 1----4GlcNac beta 1----3Gal beta 1----4GlcNac beta 1----3Gal beta 1----4GlcNAc beta 1----3Gal beta 1----4Glc-Cer. Several species of neutral glycosphingolipids with six to more than ten monosaccharides were detected which carry the X antigen and others the Y antigen: Gal beta 1----4(Fuc alpha 1----3)GlcNAc and Fuc alpha 1----2Gal beta 1----4(Fuc alpha 1----3)GlcNAc, respectively. In addition, three new types of carbohydrate specificities were detected among the myeloid cell glycolipids. Two were associated with neutral glycolipids: the first, recognised by anti-myeloid antibodies VIM-1 and VIM-10, was expressed on a distinct set of glycolipids with six or more monosaccharides, and the second, recognized by VIM-8, was expressed on glycolipids with more than ten monosaccharides. The third specificity, recognised by the anti-myeloid antibody VIM-2, was expressed on slow migrating sialoglycolipids with backbone structures of the poly-N-acetyllactosamine type that are susceptible to degradation with endo-beta-galactosidase. Thus, we conclude that the i and Y antigens occur among the glycolipids of normal myeloid and chronic myelogenous leukemia cells and that a high proportion of hybridoma antibodies raised against differentiation antigens of myeloid cells are directed at carbohydrate structures.
选用了六种对碳水化合物抗原i、X或Y具有已知特异性的单克隆抗体，以及七种因其与人白细胞不同反应模式而挑选出的抗髓细胞抗体（决定簇未知），在色谱结合试验中检测它们与源自正常人粒细胞和慢性粒细胞白血病细胞的髓细胞糖脂的反应。抗原性仅存在于微量糖脂上，用苔黑酚 - 硫酸染色几乎检测不到或根本检测不到。其中，一种中性糖鞘脂与抗 - i抗体Den结合，色谱分析显示为神经酰胺八糖，即Galβ1→4GlcNacβ1→3Galβ1→4GlcNacβ1→3Galβ1→4GlcNAcβ1→3Galβ1→4Glc - Cer。检测到几种含有六个至十多个单糖的中性糖鞘脂，其中一些带有X抗原，另一些带有Y抗原：分别为Galβ1→4(Fucα1→3)GlcNAc和Fucα1→2Galβ1→4(Fucα1→3)GlcNAc。此外，在髓细胞糖脂中还检测到三种新的碳水化合物特异性类型。两种与中性糖脂相关：第一种由抗髓细胞抗体VIM - 1和VIM - 10识别，在一组独特的含有六个或更多单糖的糖脂上表达；第二种由VIM - 8识别，在含有十多个单糖的糖脂上表达。第三种特异性由抗髓细胞抗体VIM - 2识别，在具有多聚N - 乙酰乳糖胺类型主链结构且易被内切β - 半乳糖苷酶降解的慢迁移唾液酸糖脂上表达。因此，我们得出结论，i和Y抗原存在于正常髓细胞和慢性粒细胞白血病细胞的糖脂中，并且针对髓细胞分化抗原产生的杂交瘤抗体中有很大一部分针对的是碳水化合物结构。
