Use of monoclonal antibodies against surface antigens of germ cells

近年来，对哺乳动物生殖细胞受精、增殖和分化的分子机制的研究取得了很大进展。分离Y或X精子的尝试并不完全成功，尽管已经开发出可以完全分离Y和X精子的方法；这种方法对家畜生产有很大的有利影响。这项研究的第一个目的是建立杂交瘤细胞，产生识别Y或X精子上特定表面抗原的单抗。第二个目的是阐明这些抗原的结构和生理功能。用从牛睾丸制备的成熟精子免疫小鼠，制备产生识别这些抗原的抗体的杂交瘤细胞。三种单抗与牛精子特异结合，但不与其他细胞结合。它们不仅与牛精子相互作用，还与小鼠和大鼠精子相互作用。他们识别特定的区域；第一个抗体分别与顶体突起结合，第二个抗体与中段结合，第三个抗体与尾巴结合。研究了这些抗体对精子生育力的影响。从早熟小鼠和大鼠的睾丸中提取细胞免疫小鼠，并制备了识别这些抗原的杂交瘤细胞。在精原细胞、精细胞、支持细胞和间质细胞等多种睾丸细胞上均有特异性结合的单抗。对这些抗原的鉴定以及精子抗原在精子发生过程中的表达进行了研究。有人试图克隆其中一些抗原的cDNA。

Recently, a great advance has been made in understanding molecular mechanisms of fertilization, proliferation and differentiation of mammalian germ cells. An attempt to separate Y or X sperm has not been fully successful, although development of the method by which Y and X sperms can be completely fractionated has been aspired ; such method has a great beneficial effect on production of livestock. The first purpose of this research is to establish hybridoma cells producing monoclonal antibodies that recognize specific surface antigens on Y or X sperm. The second aim is to elucidate structures and physiological functions of these antigens.Mice were immunized with matured sperms prepared from bovine testis and hybridoma cells producing antibodies that recognize the antigens, were prepared. Three monoclonal antibodies bound specifically with bovine sperm but not with other cells. They interacted not only with bovine sperm but also with mouse and rat sperms. They recognized specific regions ; the first antibody bound to the acrosoihal process, the second, to the midpiece, and the third, to the tail, respectively. Effects of these antibodies on fertility of sperms were investigated.Cells prepared from testes of prematured mice and rats gave to mice for immunization and hybridoma cells that recognize the antigens were prepared. There were monoclonal antibodies bound specifically to a various kinds of cells that are present in testes such as spermatogonia, spermatids, Sertoli cell, and Leydig cells. Identification of these antigens, and expression of the sperm antigens on spermatogenesis were investigated. An attempt to clone CDNA of some of these antigens has been made.