【摘要】：目的 免疫避孕與免疫不孕是生殖研究領(lǐng)域的兩大重要課題，尋找合適的精子膜抗原是解決這兩個問題的關(guān)鍵所在。精子膜抗原與受精和胚胎的早期發(fā)育有密切關(guān)系，其中部分抗原能引起機體產(chǎn)生相應(yīng)的抗體，導(dǎo)致免疫性不孕。 PH-20(posterior head 20)作為傳統(tǒng)的精子膜抗原，是由磷脂酰肌醇(glycophosphatidylinositol，GPI)錨定的膜結(jié)合型透明質(zhì)酸酶，最適PH值呈中性，它分布于精子表面漿膜和頂體內(nèi)膜，是一種透明質(zhì)酸酶誘導(dǎo)信號轉(zhuǎn)導(dǎo)的受體和識別透明帶的受體。最初的研究發(fā)現(xiàn)，用豚鼠PH-20免疫雌雄兩種豚鼠，獲得了100％的避孕效果。接受免疫的25只雌性豚鼠全部產(chǎn)生了高滴度抗體，專門識別精液中的PH-20和阻止精子與卵透明帶結(jié)合。但這些豚鼠在免疫后的6-15個月又逐漸恢復(fù)了生育能力。由此可知，豚鼠PH-20免疫豚鼠可得到明顯、持久但又可逆的抗生育效應(yīng)。PH-20在精子表面的分布及其在受精過程中的作用，使其作為免疫避孕候選疫苗一直受到人們的關(guān)注。 DNA疫苗是二十世紀(jì)九十年代發(fā)展起來的一種新型疫苗，它是指將編碼特定抗原的基因克隆到合適的質(zhì)粒載體上，制備成核酸表達(dá)載體，通過肌肉注射等方法將其導(dǎo)入機體內(nèi)，通過宿主細(xì)胞的轉(zhuǎn)錄系統(tǒng)合成抗原蛋白質(zhì)，，從而激發(fā)機體產(chǎn)生特異性的免疫應(yīng)答，所以又被稱為核酸疫苗或基因疫苗。DNA疫苗具有免疫持續(xù)時間長、制備、攜帶方便、廉價等優(yōu)點，它在免疫避孕方面的應(yīng)用促進(jìn)了避孕疫苗的發(fā)展。本實驗應(yīng)用RT-PCR方法擴(kuò)增出人PH-20(hPH-20)基因片段，與真核
[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.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R392

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