Saji F, Ohashi K, Kamiura S, Negoro T, Tanizawa O
Department of Obstetrics and Gynecology, Osaka University Medical School, Japan.
Am J Reprod Immunol Microbiol. 1988 Aug;17(4):128-33. doi: 10.1111/j.1600-0897.1988.tb00216.x.
A sperm antigen has been isolated from radiolabeled human sperm cell membrane by detergent solubilization, lectin affinity chromatography, gel filtration, and indirect immune precipitation using sperm-immobilizing antisera from patients with unexplained infertility. Isolated material was characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Among 20 infertile women's sera with sperm-immobilizing antibodies, two were found to react predominantly with a sperm membrane polypeptide having the approximate molecular weight of 15,000 daltons. No significant binding to this molecule was observed in any sera from pregnant women, unmarried women, and normal men. By the absorption with spermatozoa, the antisera lost their binding activity to the molecule, while the sera absorbed with seminal plasma did not lose the activity. The results indicated that the molecule is a genuine sperm antigen and not a sperm-coating seminal plasma antigen. By the indirect immunofluorescence of washed ejaculated spermatozoa with the antisera, strong fluorescence was localized only in an equatorial segment of the acrosome, while no specific staining was observed in the controls. The antigen is relatively unstable against acid, alkali, and heat treatment. Treatment with proteolytic enzymes such as pronase and trypsin inactivated the antigen activity, indicating that the antigen epitope could be a peptide portion of the glycoprotein.
通过去污剂增溶、凝集素亲和层析、凝胶过滤以及使用不明原因不孕症患者的精子制动抗血清进行间接免疫沉淀，从放射性标记的人精子细胞膜中分离出一种精子抗原。分离出的物质通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）进行表征。在20名具有精子制动抗体的不育女性血清中，发现有两份血清主要与一种分子量约为15,000道尔顿的精子膜多肽发生反应。在孕妇、未婚女性和正常男性的任何血清中均未观察到与该分子的明显结合。用精子吸收后，抗血清失去了与该分子的结合活性，而用精浆吸收的血清则未失去该活性。结果表明该分子是一种真正的精子抗原，而不是精子包被的精浆抗原。用抗血清对洗涤后的射出精子进行间接免疫荧光检测，强荧光仅定位在顶体的赤道段，而在对照中未观察到特异性染色。该抗原对酸、碱和热处理相对不稳定。用链霉蛋白酶和胰蛋白酶等蛋白水解酶处理会使抗原活性失活，表明抗原表位可能是糖蛋白的肽段部分。
