Symington F W, Hedges D L, Hakomori S
J Immunol. 1985 Apr;134(4):2498-506.
Glycolipid and cell surface carbohydrate antigens of human polymorphonuclear neutrophils (PMN) and of HL-60 myeloid leukemia cells were analyzed with a panel of defined, monoclonal anti-carbohydrate antibodies. Antigenicities of intact PMN, HL-60, and retinoic acid-induced HL-60 (r.a.-HL-60) were studied by flow cytofluorometry. These three cell populations displayed quantitative differences, some of which were induction dependent, in their expression of lactosyl, N-acetyllactosaminyl, Y-hapten (Fuc alpha 1----2Gal beta 1----4(Fuc alpha 1----3)GlcNAc beta 1----R), and sialosyl-X-hapten (SA alpha 2----3Gal beta 1----4(Fuc alpha 1----3)GlcNAc beta 1----R) specificities. Structures reactive with antibodies specific for long-chain mono-, and di- or tri- alpha 1----3 fucosylated lacto-series glycolipids were also detected. Glycosphingolipids purified from organic extracts of these cells were analyzed to seek information concerning the chemical basis for these surface antigenic differences, to assess the structural and antigenic diversity of PMN and HL-60 glycolipids, and to quantitate chemically and antigenically prominent glycolipids. Binding of monoclonal antibodies to thin-layer chromatograms demonstrated that each of the specificities on intact cells was carried by one or more distinct glycolipids. The abundance of immunoreactive glycolipids in the extracts paralleled the relative staining intensities of the intact cell populations. Several "cryptic" glycolipid antigens, including alpha 2----6 sialosylated structures enriched five- to 10-fold in PMN extracts, were not detected on intact cells. Lactosylceramide accounted for two-thirds of the approximately 1.5 X 10(9) glycolipid molecules contained in each PMN. The remaining glycolipid antigens appeared to include structurally diverse fucolipids, fucogangliosides, and neutral and sialosylated glycolipids with Gal beta 1----4GlcNAc beta 1----R terminal core structure. The abundance, diversity, and induction-dependent expression of these structures suggest that they may participate in PMN maturation and function.
利用一组经过鉴定的单克隆抗碳水化合物抗体，对人多形核中性粒细胞（PMN）和HL - 60髓系白血病细胞的糖脂及细胞表面碳水化合物抗原进行了分析。通过流式细胞荧光术研究了完整PMN、HL - 60及维甲酸诱导的HL - 60（r.a.-HL - 60）的抗原性。这三种细胞群体在乳糖基、N - 乙酰乳糖胺基、Y - 半抗原（Fucα1----2Galβ1----4（Fucα1----3）GlcNAcβ1----R）和唾液酸基 - X - 半抗原（SAα2----3Galβ1----4（Fucα1----3）GlcNAcβ1----R）特异性的表达上表现出数量差异，其中一些差异是诱导依赖性的。还检测到了与针对长链单、二或三α1----3岩藻糖基化乳糖系列糖脂特异性抗体反应的结构。对从这些细胞的有机提取物中纯化的糖鞘脂进行分析，以获取有关这些表面抗原差异化学基础的信息，评估PMN和HL - 60糖脂的结构和抗原多样性，并对化学和抗原上突出的糖脂进行定量。单克隆抗体与薄层色谱图的结合表明，完整细胞上的每种特异性由一种或多种不同的糖脂携带。提取物中免疫反应性糖脂的丰度与完整细胞群体的相对染色强度平行。在完整细胞上未检测到几种“隐蔽”的糖脂抗原，包括在PMN提取物中富集了5至10倍的α2----6唾液酸基化结构。乳糖神经酰胺占每个PMN中约1.5×10⁹个糖脂分子的三分之二。其余的糖脂抗原似乎包括结构多样的岩藻糖脂、岩藻糖神经节苷脂以及具有Galβ1----4GlcNAcβ1----R末端核心结构的中性和唾液酸化糖脂。这些结构的丰度、多样性和诱导依赖性表达表明它们可能参与PMN的成熟和功能。
