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抗人受精抗原-1(FA-1)单克隆抗体抑制牛的体外受精:在免疫避孕中的应用。

Monoclonal antibody to human fertilization antigen-1 (FA-1) inhibits bovine fertilization in vitro: application in immunocontraception.

作者信息

Coonrod S A, Westhusin M E, Naz R K

机构信息

Department of Veterinary Physiology and Pharmacology, Texas A&M University, College Station 77843.

出版信息

Biol Reprod. 1994 Jul;51(1):14-23. doi: 10.1095/biolreprod51.1.14.

Abstract

A monoclonal antibody (mAb) to the human sperm plasma membrane protein, fertilization antigen-1 (FA-1), was tested for its reactivity with bovine spermatozoa and its effects on bovine fertilization in vitro. Western blot analysis revealed that the FA-1 mAb reacted with proteins of similar molecular mass (53 +/- 2 kDa) in human and bovine sodium deoxycholate (DOC)-solubilized sperm extracts. Indirect immunofluorescence, using epifluorescence microscopy and laser scanning confocal microscopy, revealed that the FA-1 antigen is present in the post-acrosomal region of bovine spermatozoa, which is similar to human FA-1 localization. In bovine in vitro fertilization (IVF) trials, using oocytes obtained from slaughterhouse ovaries, addition of 20, 40, or 80 micrograms/ml of FA-1 mAb to the IVF medium resulted in a linear decrease in the fertilization rate from 86.3% in the controls to 54.6%, 21.6%, and 1.8% in the respective experimental groups (p < 0.01). There was no inhibitory effect (p > 0.10) of the FA-1 mAb on percent sperm motility or other motility characteristics tested, suggesting that human FA-1 mAb inhibits bovine sperm cell function at some point after capacitation. In conclusion, the evolutionarily conserved antigen FA-1 has a molecular identity in bovine sperm similar to that in human sperm, and mAb to human sperm FA-1 inhibits fertilization of bovine oocytes. These results indicate that FA-1 is a promising candidate for the development of a contraceptive vaccine. The research also suggests that bovine species could be used as a model for investigating the use of FA-1 as an immunovaccine in ruminants.

摘要

对一种针对人类精子质膜蛋白受精抗原-1(FA-1)的单克隆抗体(mAb)进行了检测,观察其与牛精子的反应性及其对牛体外受精的影响。蛋白质免疫印迹分析显示,FA-1单克隆抗体与经脱氧胆酸钠(DOC)溶解的人及牛精子提取物中分子量相似(53±2 kDa)的蛋白质发生反应。利用落射荧光显微镜和激光扫描共聚焦显微镜进行的间接免疫荧光显示,FA-1抗原存在于牛精子的顶体后区域,这与人FA-1的定位相似。在牛体外受精(IVF)试验中,使用从屠宰场卵巢获得的卵母细胞,向IVF培养基中添加20、40或80微克/毫升的FA-1单克隆抗体,导致受精率从对照组的86.3%呈线性下降,在各实验组中分别降至54.6%、21.6%和1.8%(p<0.01)。FA-1单克隆抗体对所检测的精子活力百分比或其他活力特征没有抑制作用(p>0.10),这表明人FA-1单克隆抗体在获能后的某个时间点抑制牛精子细胞功能。总之,进化上保守的抗原FA-1在牛精子中的分子特性与人精子相似,并且针对人精子FA-1的单克隆抗体抑制牛卵母细胞的受精。这些结果表明,FA-1是开发避孕疫苗的一个有前景的候选物。该研究还表明,牛物种可作为研究将FA-1用作反刍动物免疫疫苗的模型。

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