Zhang Q, Young T F, Ross R F
Veterinary Medical Research Institute, Iowa State University, Ames 50011.
Infect Immun. 1994 Oct;62(10):4367-73. doi: 10.1128/iai.62.10.4367-4373.1994.
Glycolipid receptors for Mycoplasma hyopneumoniae attachment were analyzed by using a thin-layer chromatography (TLC) overlay assay. M. hyopneumoniae bound specifically to sulfatide, globoside, and monosialoganglioside GM3. No binding to sphingomyelin, cerebroside, lactosyl ceramide, ceramide trihexoside, monosialogangliosides GM1 and GM2, disialogangliosides (GD1a, GD1b, and GD3), trisialoganglioside (GT1b), cholesterol, cholesterol sulfate, palmitic acid, tripalmitin, or cholesteryl palmitate was detected. Total lipids extracted from cilia of the swine respiratory epithelium, the natural targets of M. hyopneumoniae infection, were also separated on TLC plates and overlaid with mycoplasmas. M. hyopneumoniae bound specifically to three ciliary glycolipids identified as La, Lb, and Lc. Binding to Lc was stronger than to La and Lb. All three lipids were believed to be sulfated glycolipids, as determined by laminin binding and staining with azure A. Lc was identified as a putative sulfatide because it has a mobility similar to that of authentic sulfatide and comigrated with sulfatide on TLC plates. Laminin bound to La, Lb, and Lc and produced dose-dependent inhibition of adherence of the mycoplasma to the three ciliary receptors. Binding of the mycoplasma to sulfatide, La, Lb, and Lc was partially inhibited by dextran sulfate, heparin, fucoidan, mucin, and chondroitin sulfate B. These substances blocked the adherence of M. hyopneumoniae to cilia and ciliated cells as shown in a previous study (Q. Zhang, T. F. Young, and R. F. Ross, Infect. Immun. 62:1616-1622, 1994). These results indicate that La, Lb, and Lc are the major native receptors for M. hyopneumoniae adherence to ciliated cells.
通过使用薄层层析（TLC）覆盖分析来分析猪肺炎支原体附着的糖脂受体。猪肺炎支原体特异性结合硫脂、球苷脂和单唾液酸神经节苷脂GM3。未检测到与鞘磷脂、脑苷脂、乳糖基神经酰胺、神经酰胺三己糖苷、单唾液酸神经节苷脂GM1和GM2、双唾液酸神经节苷脂（GD1a、GD1b和GD3）、三唾液酸神经节苷脂（GT1b）、胆固醇、硫酸胆固醇、棕榈酸、三棕榈精或胆固醇棕榈酸酯的结合。从猪呼吸道上皮纤毛中提取的总脂质（猪肺炎支原体感染的天然靶标）也在TLC板上进行分离，并用支原体覆盖。猪肺炎支原体特异性结合三种鉴定为La、Lb和Lc的纤毛糖脂。与Lc的结合强于与La和Lb的结合。通过层粘连蛋白结合和天青A染色确定，所有三种脂质均被认为是硫酸化糖脂。Lc被鉴定为假定的硫脂，因为它在TLC板上具有与 authentic硫脂相似的迁移率并与硫脂共迁移。层粘连蛋白与La、Lb和Lc结合，并对支原体与三种纤毛受体的粘附产生剂量依赖性抑制。硫酸葡聚糖、肝素、岩藻依聚糖、粘蛋白和硫酸软骨素B部分抑制了支原体与硫脂、La、Lb和Lc的结合。如先前研究（Q. Zhang、T. F. Young和R. F. Ross，Infect. Immun. 62:1616 - 1622，1994）所示，这些物质阻断了猪肺炎支原体与纤毛和纤毛细胞的粘附。这些结果表明，La、Lb和Lc是猪肺炎支原体粘附纤毛细胞的主要天然受体。
