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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.

我们证明了从一名免疫性不育妇女获得的血清可以诱导人类精子的头对头凝集，并与人类精子蛋白相互作用。从人类身上收集足够数量的精液是一项艰巨的任务。为了绕过这一供应问题，我们决定研究千叶县和福冈县家畜实验站和国家畜牧业研究所供应充足的猪精子。该血清还能诱导射出的猪精子发生头对头的凝集。用间接免疫荧光法测定病人制备的免疫球蛋白G(Ig G)定位于精子头部顶体区，取自附睾体和附睾尾。通过SDS-PAGE免疫印迹技术测定，该免疫球蛋白与猪精子蛋白相互作用，估计其相对分子质量为45 kDa。然而，从从睾丸和附睾头或非性腺组织中提取的精子中提取的蛋白质不与免疫球蛋白相互作用。这种免疫球蛋白与附睾体和附睾尾液中存在的约75 kDa和38 kDa的额外蛋白质相互作用，而不与附睾头液体中的蛋白质相互作用。相互作用的蛋白在刀豆蛋白A琼脂糖柱上被吸附，表明它们是糖蛋白。研究猪精子抗原作为免疫原对实验动物生育力的影响是开发抗生育疫苗的一个有前途的方法。我们已研制出抗猪精子蛋白的单抗，可在体外诱导猪精子凝集。