Jiang H, Pillai S
Department of Obstetrics and Gynecology, Medical University of South Carolina, Charleston 29425, USA.
Am J Reprod Immunol. 1998 Apr;39(4):243-8. doi: 10.1111/j.1600-0897.1998.tb00360.x.
To determine whether complement regulatory proteins are present on human spermatozoa and whether antibodies to these proteins adversely affect sperm motility.
Human sperm membrane proteins were solubilized and subjected to polyacrylamide gel electrophoresis followed by Western blot analysis against antibodies to complement component 1 inhibitor (C1-INH), decay-activating factor (DAF; CD55), membrane cofactor protein (MCP; CD46), and homologous restriction factor (HRF; CD59). Spermatozoa, obtained by a swim-up technique, were incubated in medium (control 1) and medium supplemented with antibodies to human albumin (control 2) and antibodies to these complement regulatory proteins. We used a computerized sperm motion analysis to determine the effect of these antibodies on sperm motion characteristics.
Complement regulatory proteins such as C1-INH, CD55, CD46, and CD59 were found in the sperm extracts as shown by Western blot analysis. The heat-treated (56 degrees C, 30 min) IgG fraction of antibodies to these proteins significantly reduced sperm motility in general and other motion parameters. Addition of complement did not affect these results except in the antibodies to CD46 in which the reducing action was further amplified.
Our data suggest that C1-INH, CD55, CD46, and CD59 are present on the sperm surface. These proteins may have biological functions, such as affecting sperm motility, besides the complement regulatory functions. In infertile men and women with antibodies that recognize one or more of these complement regulatory proteins, there may be problems related to poor sperm motility and survival in the reproductive tracts.
确定补体调节蛋白是否存在于人类精子上,以及针对这些蛋白的抗体是否会对精子活力产生不利影响。
将人类精子膜蛋白溶解,进行聚丙烯酰胺凝胶电泳,然后针对补体成分1抑制剂(C1-INH)、衰变加速因子(DAF;CD55)、膜辅因子蛋白(MCP;CD46)和同源限制因子(HRF;CD59)的抗体进行蛋白质印迹分析。通过上游技术获得的精子在培养基(对照1)以及补充了抗人白蛋白抗体(对照2)和这些补体调节蛋白抗体的培养基中孵育。我们使用计算机化的精子运动分析来确定这些抗体对精子运动特征的影响。
蛋白质印迹分析显示,在精子提取物中发现了补体调节蛋白,如C1-INH、CD55、CD46和CD59。这些蛋白的抗体经热处理(56摄氏度,30分钟)的IgG部分总体上显著降低了精子活力以及其他运动参数。除了针对CD46的抗体中还原作用进一步增强外,添加补体并不影响这些结果。
我们的数据表明,C1-INH、CD55、CD46和CD59存在于精子表面。这些蛋白除了具有补体调节功能外,可能还具有生物学功能,如影响精子活力。在识别这些补体调节蛋白中的一种或多种的抗体阳性的不育男性和女性中,可能存在与生殖 tract 中精子活力差和存活率低相关的问题。 (注:原文中“reproductive tracts”直译为“生殖道”,这里的“tract”复数形式可能表述不准确,推测原文想表达的是“生殖系统”,但按照要求未做修改。)