Dubova-Mihailova M, Komori S, Kameda K, Tsuji Y, Koyama K, Isojima S
Department of Obstetrics and Gynecology, Hyogo Medical College, Japan.
J Reprod Immunol. 1994 Mar;26(2):97-110. doi: 10.1016/0165-0378(94)90034-5.
A monoclonal antibody, MAb 1G1, possessing a strong fertilization-blocking activity was prepared by immunizing a BALB/c mouse with capacitated human sperm in order to study the molecular nature of sperm antigens relevant to fertilization. MAb 1G1 inhibited human sperm penetration into zona-free hamster eggs. It reacted to the apical portion of acrosome-reacted human sperm, but did not react to live sperm before the acrosome reaction as demonstrated by immunofluorescence staining. In paraffin-embedded testis sections, the round spermatids, spermatocytes and spermatozoa were stained with MAb 1G1, but the spermatogonia were not stained. Neither Sertoli cells, Leydig cells nor other somatic tissues were stained. The sperm of Japanese monkey, bull, boar, hamster and mouse were not stained. Therefore, the staining of sperm was species specific. The antigen corresponding to MAb 1G1 showed a band at 27 kDa by immunoblotting. The reactivity of the antigenic component was not destroyed by periodic acid treatment. From the results obtained, it was postulated that the antigenic molecule might be a polypeptide. These results indicated that this MAb might be a useful tool for studying the mechanism of human sperm-egg fertilization and the development of a contraceptive vaccine.
为研究与受精相关的精子抗原的分子本质,用获能的人精子免疫BALB/c小鼠,制备了具有强受精阻断活性的单克隆抗体MAb 1G1。MAb 1G1抑制人精子穿透去透明带仓鼠卵。免疫荧光染色显示,它与顶体反应后的人精子顶端部分发生反应,但不与顶体反应前的活精子发生反应。在石蜡包埋的睾丸切片中,圆形精子细胞、精母细胞和精子用MAb 1G1染色,但精原细胞未被染色。支持细胞、间质细胞及其他体细胞均未被染色。日本猴、公牛、公猪、仓鼠和小鼠的精子未被染色。因此,精子的染色具有种属特异性。通过免疫印迹法,与MAb 1G1对应的抗原在27 kDa处显示一条带。抗原成分的反应性未被高碘酸处理破坏。根据所得结果推测,抗原分子可能是一种多肽。这些结果表明,该单克隆抗体可能是研究人类精卵受精机制和开发避孕疫苗的有用工具。