摘要
为了测定抗精子ＩｇＡ在免疫不育和抗精子避孕疫苗研制方面的生物学作用，用肠道内免疫的方法制备了一组抗乳酸脱氢酶Ｃ４（ＬＤＨ－Ｃ４）的单克隆ＩｇＡ抗体（ｍｏＩｇＡ）。以免疫印迹证实了它们的异质同形体。大部分ｍｏＩｇＡ（ＰＡ１－ＰＡ５）是用肠道内免疫和以派依尔氏淋巴细胞作为亲本细胞进行融合来获得的。在豚鼠血清补体存在的情况下，小鼠精子可以被ｍｏＩｇＡＰＡ１、ＰＡ２和ＰＡ４所制动。高浓度ＰＡ４和ＰＡ５可凝集小鼠精子。小鼠体外受精率可被３个ｍｏＩｇＡ（ＰＡ２、ＰＡ３和ＰＡ４）显著降低，但用ＰＡ１、ＰＡ２和ＰＡ５被动免疫之后，小鼠体内受精无明显变化。纯化的小鼠胆汁分泌片可同纯化的ｍｏＩｇＡ或腹水中的ｍｏＩｇＡ在体外组装起来。同分泌片结合之后，ｍｏＩｇＡ对精子的制动、凝集和体外受精无明显变化。这些研究结果提供了抗ＬＤＨ－Ｃ４的ｍｏＩｇＡ和分泌性ＩｇＡ对精子功能和体外受精的生物学作用的直接证据，在免疫不育的防治，避孕疫苗的研制以及性传播疾病的防治方面均有一定的指导意义。
in order to determine the biological role of antisperm IgA, especially secretory IgA,in immunological infertility and in the development of an antisperm contraceptive vaccine, a panel of monoclonal IgA(moIgA) antibodies to lactate dehydrogenase C4(LDH-C4) were prepared by intraintestinal immunization, and their heterogeneous isoforms were demonstrated by Western blotting. Most moIgAs(PA1-PA5) were obtained by the combined use of intraintestinal immunization and fusion of Peyer's pateh cells for parent cells.In the presence of guinea pig serum complement,mouse sperm were immobilized by moIgA PA1, PA2 and PA4. Mouse sperm were agglutinated by PA4 and PA5 at high concentration.The rates of in vitro fertilization were significantly reduced by 3 of 7 moIgA tested,but moIgA (PA1, PA2 and PA5) had no significant effects on mouse in vivo fertilization by passive immunization. Purified murine bile secretory component was assembled with either purified moIgA or moIgA in ascites in viyro.The effects of moIgA on sperm immobilization,sperm agglutination and in vitro fertilization were not significantly changed following the binding of the moIgA with secretory component.These results provide direct evidence of a positive biological role of anti-LDH-C4 moIgA and secretory IgA in sperm function and iv vitro fertilization.
关键词
单克隆抗体
免疫球蛋白
乳酸脱氢酶
Monoclonal IgA
Lactate dehydrogenase C4
Sperm immobilization and agglutination
In vitro fertilization
Secretory component
