中国科学院机构知识库网格
Chinese Academy of Sciences Institutional Repositories Grid
抗小鼠乳酸脱氢酶C4(LDH-C4)IgA单克隆抗体的制备及其对精子功能的影响

文献类型:学位论文

作者封树林
学位类别硕士
答辩日期1990
授予单位中国科学院研究生院
授予地点北京
导师贲昆龙
关键词乳酸脱氢酶C4 精子
中文摘要乳酸脱氢酶C4 (LDH-C4)是一种人和哺乳动物精子特有的乳酸脱氢酶同功酶。用纯化的小鼠LDH-C4 免疫动物,有一定的避孕效果。这种避孕效果与血清特异性抗体水平并不完全一致。这可能是由于受精过程是在生殖道内进行的,而生殖道内又有粘膜免疫因素存在。IgA抗体是生殖道内的主要抗体成分,研究抗LDH-C4 IgA抗体的抗精子作用有助于了解局部分泌性免疫系统在抗精子免疫避孕中的作用。由于足够量的特异性IgA抗体难于从动物或人粘膜分泌液中分离到,为了获得供体内外试验用的该种抗体,直接证明它在抗生育方面的作用,我们采用一种特殊的免疫方法制备了一系列的抗LDH-C4的单克隆抗体,包括6株IgA和9株IgM。这种免疫方法的主要特点是将抗原直接注射到派伊尔氏淋巴小结(PP)或小肠腔内。ELISA检测表明,按这种方法免疫后,分泌IgA的克隆出现的比例明显高于常规免疫的结果。这是因为PP是粘膜免疫的中枢,其中含有大量的IgA前体细胞,直接将抗原注射到PP内有助于刺激IgA前体细胞的分化和增殖,诱导局部分泌性免疫反应。我们用所得到的单克隆抗体研究了LDH-C4在人,小鼠和树鼩精子表面的定位。结果表明,大多数单克隆抗体可以结合到这些精子的表面;不同的单抗在同一物种的精子表面呈现不同的结合区域。这一方面说明来源于人,小鼠和树鼩的LDH-C4的抗原决定簇有很高的同源性;另一方面提示LDH-C4在精子表面不同的区域所暴露的抗原决定簇不同。初步的功能试验表明,某些抗LDH-C4的IgA单克隆抗体可以凝集或制动精子,说明生殖道内的IgA抗体可以通过凝集和制动作用来影响精子的功能,从而影响精子的受精力。
英文摘要Lactic dehydrogenase C4 (LDH-C4) is a sperm specific lactic dehydrogenase in mammals and human. Significant reduction in fertility was observed in female mice, rabbits and baboons immunized with purified mouse LDH-C4. Interestingly, the serum antibody level is not in parallel with the suppression of fertility in these animals, perhaps because that the immune system-sperm interaction site is in the reproductive tract where the factor of local immunity exists. IgA is the major immunoglobulin component in the genital secretions. Research on the antisperm effects of IgA against LDH-C4 will provide us with some information on the role played by the whole mucosal immune system in antifertility. It is impossible to collect sufficient specific IgA from the mucosal secretions of human and experimental mammals to test its effects on fertility. So, in present study hybridoma technique was used to produce monoclonal IgA to murine LDH-C4. Female Balb/c mice were immunized by directly injecting purified mouse LDH-C4 into Peyer's patches(PP) and intralumen of small instestine. Totally 15 McAbs including six IgA and nine IgM have been obtained. The result suggests that immunization by directly injecting the antigen into submucosa of PP be efficient in inducing mucosal immune response. The localization of LDH-C4 on the sperm surface of three species(mouse, tree shrew and human) was investigated with indirect immunofluorescence using unfixed or fixed sperm. Most of the McAbs are cross-reactive with sperm from these species, showing that LDH-C4 of the three species are much homologous in three-dimensional structure or antigenicity. Additionally, it appears that these McAbs have different binding region on the mouse sperm surface. This phenomenon probably means that LDH-C4 is present on the sperm surface in various kinds of conformation, which are taken by concrete LDH-C4 molecules and are dependent on the region in which they reside. The epitope differences exist between LDH-C4 molecules in different region of the sperm surface, causing the variation in McAb binding pattern. The effect of six monoclonal IgAs on sperm function was studied. Sperm immobilization and agglutination effects were observed when McAbs PA1 and PA5 were added. Therefore, the IgA in the reproductive tract may influence sperm function to reduce fertility through both sperm agglutination and sperm immobilization.
语种中文
公开日期2010-10-15
源URL[http://159.226.149.42:8088/handle/152453/6230]  
专题昆明动物研究所_其他
推荐引用方式
GB/T 7714
封树林. 抗小鼠乳酸脱氢酶C4(LDH-C4)IgA单克隆抗体的制备及其对精子功能的影响[D]. 北京. 中国科学院研究生院. 1990.

入库方式: OAI收割

来源:昆明动物研究所

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