Nasal immunization with mature sperm specific antigen CD52 for developing a contraceptive vaccine

据报道，人CD 52是存在于淋巴细胞、单核细胞、男性生殖道、射精精子和精浆中的糖基磷脂酰肌醇锚蛋白。这种抗原诱导抗体，导致妇女不孕，损害精子活力。本研究作为模型实验进行，以检查小鼠mrt-CD 52是否在小鼠中产生抗体，以阐明mer-CD 52抗原导致不育，并且该抗原可用于避孕疫苗开发。[方法]取ICR小鼠输精管制备mrt-C52，与完全弗氏佐剂一起注射给雌雄C57 BL小鼠。或者，通过鼻内途径用霍乱毒素B免疫小鼠。通过ELISA、蛋白质印迹和免疫荧光染色评估抗体产生。通过精子固定试验、体外受精试验和交配试验检测抗体的生物学效应。[结果]纯化的小鼠mrt-CD 52经皮下免疫雄性和雌性小鼠，均产生自身抗体和同种抗体。所有产生的抗体均不与mrt-CD 52的肽部分反应，表明碳水化合物部分具有免疫原性。抗体在补体存在下固定小鼠精子，但不抑制体外受精或体内生育力。[讨论]提示抗mrt-C52抗体可能导致部分不孕妇女不孕。进一步的实验是必要的，以产生伊加类抗体在女性生殖道的粘膜免疫与mrt-CD 52的避孕疫苗的开发。

Human CD52 has been reported to be a glycosylphosphatidylinositol anchor protein that is present in lymphocytes, monocytes, male reproductive tracts, ejaculated sperm and seminal plasma. This antigen induces antibodies causing infertility in women and impairs sperm motility. The present study is conducted as a model experiment to examine whether mouse mrt-CD52 produces antibodies in mice to clarify mer-CD52 antigen cause infertility and also this antigen is available for contraceptive vaccine development. [Methods] mrt-C52 was prepared from ICR mouse vas deferens and injected in male and female C57BL mice with complete Freund's adjuvant. Alternatively, mice were immunized by intranasal route with cholera toxin B. Antibody production was assessed by ELISA, western blotting and immunofluorescent staining. Biological effects of the antibodies were examined by sperm immobilizing test, in-vitro fertilization and mating experiments. [Results] Male and female mice produced auto- and iso-antibodies by immunization with purified mouse mrt-CD52 by a subcutaneous route. All the produced antibodies did not react to the peptide portion of mrt-CD52, suggesting that the carbohydrate moiety is immunogenic. The antibodies immobilized mouse sperm in the presence of complement but not inhibited in-vitro fertilization or in-vivo fertility. [Discussion] The results suggested that the antibody to mrt-C52 possibly cause infertility in some infertile women. Further experiment is necessary to produce IgA class antibody in female genital tracts for developing a contraceptive vaccine by mucosal immunization with mrt-CD52.