The study on the etiological meaning of oligosaccharide residures in immunological infertility and development of treatment for that infertility.

我们利用体外成熟的人透明带精子穿透试验的方法证明了不孕妇女血清中精子固定抗体的病因意义。在本项目中，我们研究了针对这些精子固定抗体的抗原的病因学意义，特别是寡糖残基在精子-卵子相互作用中的作用。（方法）采用体外成熟人透明带精子固定化试验（SIT）和精子穿透试验（SPT）。（结果）(1)从WHO库血清和部分不孕妇女血清中分离到具有精子固定活性的高效价血清。所有采集到的血清都阻止精子穿透并附着在人类透明带上。由此证明，精子固定抗体具有阻断精子穿透带和受精的作用。(2)利用几种凝集素进行了SIT实验，结果表明，魔豆蛋白A （C…More on A）、小麦胚芽凝集素（WGA）和蓖麻凝集素（RCA）具有SIT的阻断活性。(3)我们检测了这3种凝集素在人精子和透明带上的寡糖残留。我们用荧光技术在精子和透明带表面发现了这些低聚糖的存在。(4)我们研究了将这些寡糖残基添加到经凝集素处理的无精子穿透活性的卵母细胞上，以恢复精子穿透活性。由此可见，3种凝集素中只有甘露糖有恢复精子穿透的作用。我们证明了甘露糖在精子-卵子相互作用中起重要作用。(5)另一方面，我们检测了几种低聚糖预孵育精子时精子穿透活性的变化。我们还发现，只有甘露糖具有阻断精子穿透的活性。这种活性是可逆的，与精子顶体反应无关。（结论）提示精子和卵子上的寡糖残基在精卵相互作用中起重要作用，在免疫性不孕中具有病因学意义。我们发现低聚寡糖具有治疗这种免疫性不孕症的可能性。少

We have demonstrated that the etiological meaning of sperm immobilizing antibodies in sera of infertile women using the method of sperm penetration test using human zona pellucida matured in vitro. In this project, we studied on the etiological meaning of the antigens against those sperm immobilizing antibodies, especially the role of oligosaccharide residues in sperm-egg interaction.(Methods) Sperm immobilizing test (SIT) and sperm penetration test using human zona pellucida matured in vitro (SPT) were used.(Results) (1) We picked up the high titer sera with sperm immobilizing activities by SIT from WHO bank sera and the sera of some kinds of infertile women. All of the picked up sera blocked sperm penetration and attachment to human zona pellucida. From this result, we demonstrated that sperm immobilizing antibodies have the blocking activities of sperm penetration to zona and fertilization. (2) We performed the SIT using some kinds of lectins, and demonstrated that Concanavalin A (C … More on A), Wheat germ agglutinin (WGA) and Ricinus communis agglutinin (RCA) have the blocing activities of SIT. (3) We examined the existence of the oligosaccharide residures against these 3 lectins on human sperm and zona pellucida. We found the existence of these oligosaccharides on the surface of sperm and zona pellusida by fluoresent technique. (4) We studied the recovery of sperm penetration activities by addition of these oligosaccharide residues on the oocytes that were treated by lectins and had no activity of sperm penetration. From this result, only one lectin of these 3 lectins, mannose, had the effect of recovery of sperm penetration. We demonstrated that mannose have the important role on sperm-egg interaction. (5) On the other hand, we examined the change of activity of sperm penetration when sperm were pre-incubated with some kinds of oligosaccharides. We also found that only mannose have the blocking activity of sperm penetration. This activites were reversible and were not concerned with sperm acrosome reaction.(Conclusion) These date indicated that the oligosaccharide residues on sperm and egg have the important role on sperm-egg interaction, and have the etiological meaning in immunological infertility. And we found that oligosaochride had posibility of treatment for this immunological infertility using oligosaccharides. Less