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目的:评价口服卵透明带DNA避孕疫苗pVAX1-pZP3α壳聚糖纳米微粒诱发的黏膜免疫反应、抗生育作用和对卵巢结构的影响,探讨用黏膜免疫避免卵透明带抗原免疫引起卵巢功能紊乱的可行性。方法:制备pVAX1-pZP3α壳聚糖纳米微粒,10只昆明系小鼠口服免疫,第3周和第6周分别加强免疫一次,剂量为0 5mL/只,含pVAX1-pZP3αDNA50μg;对照组10只雌小鼠口服相同剂量不含pVAX1-pZP3α的壳聚糖。免疫前和免疫后隔周取小鼠血清、阴道冲洗液,用ELISA法检测抗pZP3IgA和IgG。首次免疫后第12周雌雄合笼交配,统计雌鼠怀孕情况;解剖取出双侧卵巢,制作石蜡切片,光镜观察卵巢病理学变化。结果:免疫组80%小鼠的血清和50%小鼠的阴道冲洗液可检测到抗pZP3IgA,反应强度与持续时间有明显的个体差异;免疫组5只小鼠未怀孕,对照组全部怀孕,抗生育效果与合笼时抗pZP3IgA反应有关;免疫组未怀孕小鼠卵巢组织结构与免疫组怀孕小鼠及对照组小鼠无差别。结论:口服pVAX1-pZP3α壳聚糖纳米微粒免疫小鼠能诱发生殖道黏膜免疫反应,发挥抗生育作用,对卵巢组织结构没有影响。黏膜免疫是克服卵透明带抗原免疫引起卵巢功能紊乱的一种值得深入研究的方法。
OBJECTIVE: To evaluate the mucosal immune response, anti-fertility and ovarian structure induced by ovo zona pellucida contraceptive vaccine pVAX1-pZP3α chitosan nanoparticles and to explore the feasibility of using mucosal immunization to prevent ovarian zona pellucida antigen-induced ovarian dysfunction Sex. Methods: 10 mice were immunized orally with pVAX1-pZP3α chitosan nanoparticles and 10 Kunming mice at the third week and the sixth week respectively. The dose was 0 5 mL / mouse, containing 50 μg of pVAX1-pZP3α DNA. The control group of 10 female Mice orally administered the same dose of pVAX1-pZP3α-free chitosan. Serum and vaginal washings were taken before and after immunization, and anti-pZP3IgA and IgG were detected by ELISA. Twelve weeks after the first immunization, males and females were mated to calculate the pregnancy status of females. The ovaries were dissected and paraffin sections were made. The pathological changes of ovaries were observed with light microscope. Results: The anti-pZP3IgA was detected in 80% of the sera and 50% of the vaginal washes in the immunized group. There was a significant individual difference in the intensity and duration of the reaction. Five mice in the immunized group were not pregnant and all the pregnant women in the control group were pregnant. The anti-fertility effect was related to the anti-pZP3IgA response when the cage was closed. The ovarian tissue structure of the non-pregnant mice in the immunized group was not different from that of the immunized group. Conclusion: Oral administration of pVAX1-pZP3α chitosan nanoparticles can induce reproductive tract mucosal immune response and exert anti-fertility effects on ovarian tissue structure. Mucosal immunity is a method worthy of further study to overcome ovarian dysfunction caused by immunization of zona pellucida antigen.