人精子抗原决定簇的末端单糖参与小鼠IgA单抗与抗原的结合  

THE TERMINAL MONOSACCHARIDE OF ANTIGENIC DETERMINANT IS INVOLVED IN THE BINDING OF HUMAN SPERM TO SPECIFIC MONO-CLONAL IGA ANTIBODY

作　　者：陈云亮[1] 曹筱梅[1] 

机构地区：[1]中国科学院昆明动物研究所,昆明650223

出　　处：《实验生物学报》1996年第2期101-107,共7页Acta Biologiae Experimentalis Sinica

基　　金：国家自然科学基金(39170762);云南省应用基础研究基金93C070的资助~~

摘　　要：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。In order to know whether terminal monosaccharide of human sperm antigen is involved in stimulating antisperm immune response (especialy IgA production) in mucosal immune system, 33 IgA, 12 IgG and 35 IgM monoclonal antibodies (MAbs) all obtained by intragastrointes-tinal immunization were used in the present study. The molecular weight (MW) of sperm antigens reactive with MAbs were dectected by Western blotting. The MW range of sperm antigens to the above mon-clonal IgA, IgM and IgG class, except 12 MAbs not reactive with blotted sperm antigen on nitrocellulose strip, is 10-89 KDa, 11-75KDa and .12-94 KDa respectively. The sperm antigens were separately blocked and digested with 5 lectins and 4 glycosidases. After that, the capacity of human sperm antigens bind with the above antibodies were determined by ELISA. One or more terminal monosaccharides of sperm antigens are involved with the reaction of most MAbs tested. The loss of terminal α-fucose, α-N-acetylgalactosamine, α-N-acetylglucosamine, mannose and β-galactose but neuraminic acid in the antigenic determinant of human sperm were found to seriously affect the IgA binding with the corresponding sperm antigen. The binding of IgG with sperm antigen was obviously damaged by the removing of terminal α-fucose,α-N-acetylgalactosamine and α-mannose. The results suggest that the terminal monosaccharides of the sperm antigens play an important role in stimulation of antisperm IgA and other antibodies produced by using intragastrointestinal immunization.

关 键 词：疫苗 精子抗原 免疫球蛋白A 抗体 末端单糖 

分 类 号：R392－33[医药卫生—免疫学]