Perez Martinez S, Conesa D, Cuasnicú P S
Instituto de Biología y Medicina Experimental, Buenos Aires, Argentina.
J Reprod Immunol. 1995 May;29(1):31-45. doi: 10.1016/0165-0378(95)00927-d.
Previous studies in our laboratory indicated that immunization of male and female Wistar and Lewis rats with epididymal protein DE, resulted in the development of anti-DE antibodies in over 90% of the animals, with a significant and reversible reduction of fertility. In the present study, ELISA assays performed to analyze the evolution of the immune response indicated that antibody levels in the sera of immunized animals reached a maximum at 8 weeks after the initial injection and then gradually decreased, returning to control values by the end of the sixth month. Western blot experiments demonstrated that the immune sera specifically recognized DE in epididymal sperm extracts and epididymal cytosol, while no reaction was observed with different reproductive and essential organs. The immune sera were also capable of recognizing DE on the surface of both fresh and capacitated sperm as indicated by indirect immunofluorescence experiments. Finally, the exposure of sperm to immune sera prior to uterine insemination resulted in a significant (P < 0.05) reduction in the percentage of fertilized eggs compared to controls, with no effect on sperm motility and viability, nor on their ability to undergo capacitation. Together, these results support the participation of the raised antibodies as mediators of the antifertility effect and suggest a specific interference at the sperm-egg interaction level.
我们实验室之前的研究表明,用附睾蛋白DE对雄性和雌性Wistar及Lewis大鼠进行免疫,超过90%的动物体内会产生抗DE抗体,且生育力会出现显著且可逆的下降。在本研究中,为分析免疫反应的演变而进行的ELISA检测表明,免疫动物血清中的抗体水平在初次注射后8周达到最高值,随后逐渐下降,在第六个月末恢复到对照值。蛋白质印迹实验表明,免疫血清能特异性识别附睾精子提取物和附睾细胞溶胶中的DE,而在不同的生殖器官和重要器官中未观察到反应。间接免疫荧光实验表明,免疫血清也能够识别新鲜精子和获能精子表面的DE。最后,与对照组相比,在子宫授精前让精子接触免疫血清会导致受精卵百分比显著降低(P < 0.05),且对精子活力、生存能力及其获能能力均无影响。这些结果共同支持了所产生的抗体作为抗生育作用介质的参与,并表明在精子 - 卵子相互作用水平存在特异性干扰。