人精子膜抗原PH-20 DNA疫苗载体的构建及其体内外表达

发布时间：2018-07-15 13:08

【摘要】：目的免疫避孕与免疫不孕是生殖研究领域的两大重要课题，寻找合适的精子膜抗原是解决这两个问题的关键所在。精子膜抗原与受精和胚胎的早期发育有密切关系，其中部分抗原能引起机体产生相应的抗体，导致免疫性不孕。 PH-20(posterior head 20)作为传统的精子膜抗原，是由磷脂酰肌醇(glycophosphatidylinositol，GPI)锚定的膜结合型透明质酸酶，最适PH值呈中性，它分布于精子表面浆膜和顶体内膜，是一种透明质酸酶诱导信号转导的受体和识别透明带的受体。最初的研究发现，用豚鼠PH-20免疫雌雄两种豚鼠，获得了100％的避孕效果。接受免疫的25只雌性豚鼠全部产生了高滴度抗体，专门识别精液中的PH-20和阻止精子与卵透明带结合。但这些豚鼠在免疫后的6-15个月又逐渐恢复了生育能力。由此可知，豚鼠PH-20免疫豚鼠可得到明显、持久但又可逆的抗生育效应。PH-20在精子表面的分布及其在受精过程中的作用，使其作为免疫避孕候选疫苗一直受到人们的关注。 DNA疫苗是二十世纪九十年代发展起来的一种新型疫苗，它是指将编码特定抗原的基因克隆到合适的质粒载体上，制备成核酸表达载体，通过肌肉注射等方法将其导入机体内，通过宿主细胞的转录系统合成抗原蛋白质，，从而激发机体产生特异性的免疫应答，所以又被称为核酸疫苗或基因疫苗。DNA疫苗具有免疫持续时间长、制备、携带方便、廉价等优点，它在免疫避孕方面的应用促进了避孕疫苗的发展。本实验应用RT-PCR方法扩增出人PH-20(hPH-20)基因片段，与真核
[Abstract]:Objective Immunocontraception and immune infertility are two important subjects in the field of reproductive research. Finding suitable sperm membrane antigen is the key to solve these two problems. Sperm membrane antigens are closely related to fertilization and early development of embryos, and some of them can cause the body to produce corresponding antibodies. PH-20 (posterior head 20, as a traditional sperm membrane antigen, is a membrane-bound hyaluronidase anchored by phosphatidylinositol (GPI). It is a hyaluronidase-induced signal transduction receptor and a zona-recognition receptor. The initial study found that 100% contraceptive effect was achieved by immunizing both male and female guinea pigs with PH-20. All 25 female guinea pigs immunized produced high titer antibodies that specifically identified PH-20 in semen and prevented sperm from binding to the pellucida. But the guinea pigs gradually recovered their fertility 6-15 months after immunization. It can be concluded that guinea pigs immunized with PH-20 have obvious, persistent but reversible antifertility effect. PH-20 distribution on sperm surface and its role in fertilization. DNA vaccine is a new vaccine developed in the 1990s. It refers to the cloning of genes encoding specific antigens into appropriate plasmid vectors, the preparation of nucleic acid expression vectors, and the introduction of them into the body by intramuscular injection, and the synthesis of antigen proteins through the transcription system of host cells. Therefore, it is also called nucleic acid vaccine or gene vaccine. DNA vaccine has the advantages of long immune duration, easy preparation, easy to carry, low cost and so on. Its application in the field of immunological contraception has promoted the development of contraceptive vaccine. Human PH-20 (hPH-20) gene fragment was amplified by RT-PCR.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392

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