论文部分内容阅读
精子特异性乳酸脱氢酶是精子能量代谢的一个关键酶,具有很强的组织特异性和免疫原性.在本研究中,将人和小鼠(Mus musculus)的精子特异性乳酸脱氢酶编码序列分别与真核表达载体pVAX1连接,构建了两个DNA疫苗:pVAX1-hLDHC和pVAX1-mLDHC.将这两个DNA疫苗通过黏膜表面滴注的方式分别免疫雌雄两性BALB/c小鼠,在实验小鼠的血清和雌性实验小鼠的阴道分泌液中均检测到特异性抗体的存在,滴度随免疫次数的增加而升高.交配实验中,实验组出生子代的数量明显减少,有的小鼠甚至未生育,与未免疫组和pVAX1空质粒免疫组在统计学上有显著性差异.精子凝集实验表明,实验小鼠血清和雌性实验小鼠阴道分泌液均可引起正常雄性小鼠的精子悬液发生凝集反应.免疫组化结果显示:实验小鼠血清和雌性实验小鼠阴道分泌液中的抗体与位于正常精子胞质、顶体外膜及顶体囊中的乳酸脱氢酶抗原发生特异性结合.本研究初步显示了人用避孕DNA疫苗免疫效果,为以后可能进行的临床实验奠定了基础.
Sperm-specific lactate dehydrogenase is a key enzyme of sperm energy metabolism, with strong tissue specificity and immunogenicity.In this study, human and mouse (Mus musculus) sperm-specific lactate dehydrogenase Two DNA vaccines, pVAX1-hLDHC and pVAX1-mLDHC, were constructed by ligating the two DNA vaccines with the eukaryotic expression vector pVAX1, respectively. BALB / c mice were immunized with the two DNA vaccines through mucosal instillation, The presence of specific antibodies was detected in both the serum of the experimental mice and the vaginal secretions of the female experimental mice, and the titer increased with the increase of the number of immunizations.In the mating experiment, the number of progeny in the experimental group decreased significantly Of the mice did not even have fertility and there was a statistically significant difference between the immunized group and the non-vaccinated group and the pVAX1 empty plasmid immunization group.Sperm agglutination test showed that both experimental mice serum and female experimental mice vaginal secretion can cause normal male mice Of the sperm suspension agglutination reaction.Immunohistochemical results showed that: experimental mice serum and female mice vaginal secretions test antibody and located in the normal sperm cytoplasm, the apical and acrosomal lactic acid dehydrogenation Antigen-specific binding. This preliminary study shows people with immune effect of DNA contraceptive vaccine, it laid the foundation for future clinical trials may be conducted.