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研究消化道免疫后获得的抗人精子单抗(特别是IgA单抗)的靶抗原分子性质,有助于了解哪一些抗原可以通过消化道免疫,刺激机体产生局部分泌性兔疫反应。这对于抗精子避孕疫苗研制中精子有效抗原和免疫避孕投药途径的选择,阐明精子局部免疫的发生机制和弄清免疫性不育病人的病因有一定的意义。本文对消化道免疫获得的33个抗精子IgA单抗,12个IgG和35个IgM单抗靶抗原的生化性质及末端单糖做了鉴定。免疫印迹的结果显示,IgA,IgM和IgG类单抗靶抗原的分子量范围分别为10—89,11—75和12—94 KDa。有12个单抗的靶抗原为非蛋白类的糖复合物,一个IgA单抗(A 22)的靶抗原为不含糖的蛋白。凝集素封闭和糖苷酶消化试验的结果显示,98.7%单抗的靶抗原分子末端含一种或几种糖。五种凝集素对IgA类单抗靶抗原的封闭效应均较强,表明IgA类单抗靶抗原的抗原决定簇含有岩藻糖,乙酰氨基葡萄糖,乙酰氨基半乳精,半乳糖和甘露糖等末端单糖者较多。IgG类单抗靶抗原的抗原决定簇则含有带岩藻糖,乙酰氨基半乳糖和α-甘露糖等末端单糖者较多。内切-β-半乳糖苷酶消化试验的结果表明,54.4%IgA类单抗的靶抗原为依赖半乳糖苷连接的糖肽化合物。这些结果表明,经消化道免疫能产生IgA及其它类别抗体的绝大多数人精子抗原为含多种类型末端单糖的膜表面分子。
Studying the molecular properties of target antigens of anti-human sperm monoclonal antibodies (especially IgA monoclonal antibodies) obtained after digestive tract immunization helps to understand which antigens can be immunized through the digestive tract and stimulate the body to produce a local secretory immune response. It is of some significance for the selection of sperm effective antigens and immunocontraception in the development of anti-sperm contraception vaccine, clarifying the mechanism of sperm local immunity and clarifying the etiology of immune-infertility patients. In this paper, biochemical properties and terminal monosaccharides of 33 anti-sperm IgA monoclonal antibodies, 12 IgGs and 35 IgM monoclonal antibodies obtained from digestive tract immunization were identified. The results of immunoblotting showed that the molecular weights of the target antigen of IgA, IgM and IgG monoclonal antibodies ranged from 10-89, 11-75 and 12-94 KDa, respectively. The target antigen for 12 mAbs was a non-protein carbohydrate complex and the target antigen for one IgA mAb (A 22) was a sugar-free protein. Results of lectin blocking and glycosidase digestion assays show that one or more sugars are present at the end of the target antigen molecule at 98.7% of the mAb. The blocking effects of the five lectins on the target antigens of IgA monoclonal antibodies were strong, indicating that the antigenic determinants of the IgA antigens include fucose, acetylglucosamine, acetamidogalactan, galactose, mannose and the like End monosaccharides are more. The antigenic determinants of the IgG monoclonal antibodies target more monosaccharides with terminal monosaccharides such as fucose, acetamido galactose and alpha-mannose. The results of the endo-beta-galactosidase digestion assay showed that the target antigen of 54.4% IgA monoclonal antibody was a galactoside-linked glycopeptide compound. These results indicate that most human sperm antigens that produce IgA and other classes of antibodies via digestive immunity are membrane surface molecules that contain multiple types of terminal monosaccharides.