IMMUNOLOGIC MECHANISMS IN HUMAN FEMALE INFERTILITY

患者血清、卵泡液和/或 子宫颈粘液已显示与生育力降低的大鼠相关。 该研究项目将开发一种灵长类动物模型， 精子凝集抗原-1（佐贺-1），一种精子 一种糖蛋白，已被鉴定为参与 抗体介导的人类不育。佐贺-1被鉴定为具有 鼠单克隆抗体（mAb），S19，凝集人 精子，抑制精子盲，阻止精子穿透， 子宫颈粘液，并诱发抖动现象。佐贺-1也是 由H6- 3C 4识别的人精子凝集mAb， 免疫性不孕妇女的淋巴细胞。人类H6- 3C 4 抗体和功能抑制性mAb S19识别碳水化合物 表位存在于佐贺-1糖蛋白上。微序列分析 首次证实佐贺-1与GPI锚定蛋白CDw 52相关 在淋巴细胞上鉴定并参与信号转导。S19和 H6- 3C 4碳水化合物表位不存在于CDw 252糖蛋白上。 因此，佐贺-1和CDw 52代表糖型，即糖蛋白 具有相同的肽序列但具有不同的 低聚糖侧链。本研究提案的具体目标 包括阐明佐贺-1的碳水化合物结构的研究 糖型佐贺-1 S19糖蛋白的组织特异性表达 将用免疫化学分析研究表位。推定的 佐贺-1在精子-精子/细胞-细胞外基质相互作用中的作用将 使用人半胱氨酸测定进行检查。为了研究佐贺-1作为一种 免疫性不育相关的自身、异源抗原，雌性猕猴 将用佐贺-1免疫。血清中的抗SAGA-1抗体滴度， 将监测宫颈粘液和输卵管液， 这些抗体对精子功能的影响将在体外进行测定。的 拟议的研究将有助于了解佐贺-1的作用， 精子糖萼在精子-精子/细胞-细胞外基质相互作用中作用 并将为抗体的研究提供一个非人灵长类动物模型- 介导的不育。

Titers of anti-sperm antibodies in patient sera, follicular fluid, and/or cervical mucus have been shown to correlate with reduced fertility rats. This research project will develop a primate model for immunologic infertility based on Sperm Agglutination Antigen-1 (SAGA-1), a sperm glycoprotein that has been identified as a surface antigen involved in antibody-mediated infertility in humans. SAGA-1 was identified with a murine monoclonal antibody (mAb), S19, that agglutinates human spermatozoa, inhibits sperm-zona blinding, blocks sperm penetration of cervical mucus, and induces the shaking phenomena. SAGA-1 is also recognized by H6-3C4, a human sperm-agglutinating mAb produced using lymphocytes from an immunologically infertile women. The human H6-3C4 antibody and the function-inhibitory mAb S19 recognize carbohydrate epitopes present on the SAGA-1 glycoprotein. Microsequence analysis demonstrated that SAGA-1 is related to CDw52, a GPI-anchored protein first identified on lymphocytes and involved in signal transduction. The S19 and H6-3C4 carbohydrate epitopes are not present on the CDw252 glycoprotein. Therefore, SAGA-1 and CDw52 represent glycoforms, i.e. glycoprotein isoforms that have the same peptide sequence but possess different oligosaccharide side chains. The specific aims of this research proposal includes studies to elucidate the carbohydrate structure of the SAGA-1 glycoform. The tissue-specific expression of the SAGA-1 S19 carbohydrate epitope will be investigated with immunochemical analyses. The putative role of SAGA-1 in sperm-zona/cell-extracellular matrix interactions will be examined using the human hemi-zona assay. To investigate SAGA-1 as an auto-, iso-, antigen involved in immunologic infertility, female macaques will be immunized with SAGA-1. Anti-SAGA-1 antibody titers in sera, cervical mucus, and oviductal fluid will be monitored and the effect(s) of these antibodies on sperm function will be determined in vitro. The proposed research will contribute to understanding the roles of SAGA-1 and the sperm glycocalyx in sperm-zona/cell-extracellular matrix interactions and will provide a non-human primate model for the study of antibody- mediated infertility.