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Development of monoclonal antibody for human sperm protein

人精子蛋白单克隆抗体的研制

基本信息

批准号：
05671300
负责人：
HANEJI Tatsuji
金额：
$ 1.41万
依托单位：
Kyushu Dental College
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1995
项目状态：
已结题

项目摘要

We demonstrated that serum obtained from an immunologic infertile woman induced head-to-head agglutination of human sperm and interacted with human sperm proteins. To collect sufficient amounts of semen from human beings is a difficult task. to circumvent this problem of supply, we decided to study boar sperm which is in abundant supply from the Livestock Experimental Station of Chiba and Fukuoka Prefecture, and the National Institute of Animal Industry. The serum also induced a head-to-head agglutination of ejaculated boar sperm. The immunoglobulin G (IgG) prepared from the patient localized to the acrosomal region of the sperm head obatined from the corpus and cauda epididymis as determined by an indirect immunofluorescent method. The IgG interacted with a boar sperm protein with an estimated molecular weight of 45-kDa, determined by SDS-PAGE immunoblotting technique. However, the IgG did not interact with proteins extracted from sperm obtained from the testis and caput epididymis or from non-gonadal tissues. The IgG interacted with additional proteins of about 75- and 38-kDa present in the corpus and cauda epididymal fluids but not those in the caput epididymal fluid. The interacting proteins were adsorbed when chromatographed on Concanavalin A Sepharose column, suggesting that they are glycoproteins.A promising approach in the development of an antifertility vaccine is to study the effects of the boar sperm antigens as immunogens on the fertility of experimental animals. We have developed monoclonal antibodies against boar sperm proteins that induce boar sperm agglutination in vitro.

我们证明了从免疫性不孕妇女获得的血清诱导人精子的头对头凝集，并与人精子蛋白相互作用。从人类身上收集足够数量的精液是一项艰巨的任务。为了解决这一供应问题，我们决定研究从千叶和福冈县的家畜实验站以及国家动物工业研究所大量供应的公猪精子。该血清还诱导射出的公猪精子的头对头凝集。用间接免疫荧光法测定，病人的IgG定位于附睾体和尾部精子头部的顶体区。通过SDS-PAGE免疫印迹技术测定，IgG与估计分子量为45-kDa的猪精子蛋白相互作用。然而，IgG不与从睾丸和附睾头或非性腺组织获得的精子中提取的蛋白质相互作用。IgG与附睾体和尾部液中存在的约75和38 kDa的额外蛋白质相互作用，但不与附睾头液中的蛋白质相互作用。研究猪精子抗原作为免疫原对实验动物生育力的影响，将有助于抗生育疫苗的研制。我们已经开发了抗猪精子蛋白的单克隆抗体，在体外诱导猪精子凝集。