D'Cruz O J, Lambert H, Haas G G
Department of Obstetrics and Gynecology, University of Oklahoma Health Sciences Center, Oklahoma City, USA.
Am J Reprod Immunol. 1997 Feb;37(2):172-83. doi: 10.1111/j.1600-0897.1997.tb00209.x.
The carbohydrate epitope 3-fucosyl-N-acetyllactosamine (CD15) is a constituent of cell surface glycoconjugates that has been implicated in cell-cell adhesion mediated by carbohydrate-specific ligands. The present study was designed to investigate whether CD15 is present on human sperm and whether it plays a role in human sperm-egg interaction.
Fluorescent flow cytometry was used to quantitate the binding of monoclonal antibodies (mAb) to sperm-bound CD15 and CD46 antigens on acrosome-intact (AI) and acrosome-reacted (AR) sperm. The location of the binding site of these mAbs was assessed by fluorescence microscopy. The effects of anti-CD15 and anti-CD46 mAbs on gamete interaction were tested utilizing both homologous (human zona binding and penetration) and heterologous (zona-free hamster egg binding and penetration) systems.
The mean percentage of capacitated sperm which bound anti-CD15 or anti-CD46 mAbs was low (4.8% and 5.1%, respectively). Exposure to calcium ionophore A23187 (CaI) resulted in an increase in anti-CD15 (38.6 +/- 4%) and anti-CD46 (83.4 +/- 2%) binding to sperm. Both anti-CD15 and anti-CD46 binding sites were localized by fluorescence microscopy on the sperm acrosomal region. In four experiments, the percent of zona-free hamster eggs penetrated by human sperm were medium control 93% (62/66), irrelevant mAb 74% (70/94), anti-CD46 0% (0/107), and anti-CD15 10% (9/90). One hundred percent (6/6) of human zona were penetrated by human sperm exposed to medium control, 88% (8/9) following exposure to irrelevant mAb, 0% (0/11) following exposure to anti-CD46, and 50% (5/10) following exposure to anti-CD15. The mean (+/- SD of tightly bound sperm to hamster eggs were medium control 57 +/- 18%, irrelevant mAb 64 +/- 16%, anti-CD46 37 +/- 13%, and anti-CD15 19 +/- 10%. The corresponding values for human zona were: medium control 118 +/- 14%, irrelevant mAb 61 +/- 11%, anti-CD46 39 +/- 18%, and anti-CD15 99 +/- 19%.
CD15 antigen is expressed on human sperm that have undergone acrosomal loss. mAb to CD15 was shown to inhibit significantly sperm binding and penetration of zona-free hamster eggs and penetration of human zona pellucida. These findings suggested that sperm-egg interaction may be mediated in part by the CD15 antigen. Capsule: Acrosome-reacted human sperm bind monoclonal antibodies specific for CD15 (Lewis(x)) epitope. The binding sites were located on the sperm head. Anti-CD15 antibody impaired both the binding and penetration of zona-free hamster eggs and the penetration of human zona by human sperm.
碳水化合物表位3-岩藻糖基-N-乙酰乳糖胺（CD15）是细胞表面糖缀合物的一个组成部分，与碳水化合物特异性配体介导的细胞间黏附有关。本研究旨在调查CD15是否存在于人类精子上，以及它是否在人类精卵相互作用中发挥作用。
采用荧光流式细胞术定量单克隆抗体（mAb）与顶体完整（AI）和顶体反应（AR）精子上结合的CD15和CD46抗原的结合情况。通过荧光显微镜评估这些mAb结合位点的位置。利用同源（人透明带结合和穿透）和异源（无透明带仓鼠卵结合和穿透）系统测试抗CD15和抗CD46 mAb对配子相互作用的影响。
结合抗CD15或抗CD46 mAb的获能精子的平均百分比很低（分别为4.8%和5.1%）。暴露于钙离子载体A23187（CaI）导致抗CD15（38.6±4%）和抗CD46（83.4±2%）与精子的结合增加。通过荧光显微镜观察，抗CD15和抗CD46的结合位点均位于精子顶体区域。在四项实验中，人精子穿透无透明带仓鼠卵的百分比分别为：培养基对照93%（62/66）、无关mAb 74%（70/94）、抗CD46 0%（0/107）和抗CD15 10%（9/90）。暴露于培养基对照的人精子穿透人透明带的比例为100%（6/6），暴露于无关mAb后为88%（8/9），暴露于抗CD46后为0%（0/11），暴露于抗CD15后为50%（5/10）。紧密结合到仓鼠卵上的精子的平均值（±标准差）分别为：培养基对照57±18%、无关mAb 64±16%、抗CD46 37±13%和抗CD15 19±10%。人透明带的相应值为：培养基对照118±14%、无关mAb 61±11%、抗CD46 39±18%和抗CD15 99±19%。
CD15抗原在经历顶体丢失的人类精子上表达。已证明抗CD15的mAb能显著抑制精子与无透明带仓鼠卵的结合和穿透以及人精子对人透明带的穿透。这些发现表明精卵相互作用可能部分由CD15抗原介导。总结：顶体反应的人类精子结合针对CD15（Lewis(x)）表位的单克隆抗体。结合位点位于精子头部。抗CD15抗体损害了无透明带仓鼠卵的结合和穿透以及人精子对人透明带的穿透。
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