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Distribution and role on fertilization of porcine zona pellucida antigens analyzed by monoclonal antibodies
regional staining of sperm structures was noted with different MoAb's while some MoAb's recognized similar epitopes on sperm of several species. We have identified MoAb's that inhibited human sperm from penetration of cervical mucus, zona-free animal ova or both some of which were independent of agglutinating and immobilizing activity. In the rabbit, MoAb's were found that inhibited sperm-ovum interactions as well as in vivo fertilization and in the case of two MoAb's fertilization occurred but a postfertilization infertility was noted. The latter is possibly due to some type of extra nuclear developmental signal originating on the plasma membrane of the sperm. Identity of the corresponding antigens of the MoAb's has been achieved using the Western Blot procedure. The MoAb's demonstrating antifertility effects were reactive with antigens on the plasma membrane of sperm as observed by indirect immunofluorescence. A profile of sperm antigens and their possible involvement in fertility will be useful for further delineation of immunologic infertility and development of an immunologic contraceptive.
DISTRIBUTION AND ROLE ON FERTILIZATION OF PC~CINE ZON~ PELLUCIDA ANTIGENS ANALYZED BY MONOCLONAL ANTIBODIES S.Isojima, K.Koyama, A.Hasegawa, Y.Tsunoda* and A.Hanada* Department of Obstetrics and Gynecology, Hyogo Medical College, Nishinomiya, 663 and *National Institute of Animal Industry, Tsukuba, 305, Japan Five hybridomas which produce monoclonal antibodies (Moabs) to porcine zona pellucida (Z.P.) were established. The immunoglobulin classes were IgG2a from hybridoma BIIC8 and IgM from the other four. Under immunofluorescent staining, the Moab from BIIC8 stained only porcine oocytes, the Moab from GIOG5 stained oocytes of pigs and hamsters but Moabs from C6HI, D3H4, GIOF9 stained oocytes of pigs, humans, hamsters, rats and mice. Moabs from BIIC8 and GIOF9 strongly blocked boar sperm binding to porcine Z.P. but other Moabs only did slightly. However no Moab could block the sperm penetration on in vitro fertilization (IVF) of hamster oocytes. When a second antibody was applied after each Moab, only Moab from GIOG5 impaired IVF of Hamster oocytes. Two hybridomas producing Moabs to solubilized porcine Z.P. protein which reacted only to Z.P.s of pigs and humans were also selectively established. The immunoglobulin classes from both hybridomas (3A4-2GI, IDS-2B7) were IgGl. These Moabs stained specifically Z.P. but no other tissues of pigs by immunofluorescent staining. When human oocytes were pretreated with each Moab or a combination of two Moabs, no inhibitory effects on the sperm binding to oocytes were observed, whereas the pretreatment of human oocytes with conventional mouse antiserum to porcine Z.P. blocked completely the sperm binding to oocytes. When rabbit antiserum to mouse y-globulin was added as a second antibody to the human oocy~ tes pretreated with each Moab, the spex~ binding to oocytes was completed impaired with formation of a light scattering immunoprecipitin line on the surface of Z.P.
IMMUNOHISTOLOGICAL AND SEROLOGICAL APPLICATIONS OF MONOCLONAL ANTIBODIES TO HUMAN TROPHOBLAST MEMBRANE ANTIGENS IN PREGNANCY AND MALIGNANCY P.M. JOHNSON Dept. of Irmnunology, University of Liverpool, P.O. Box 147, Liverpool. L69 3BX, England. Monoclonal antibodies (mAbs) have proven useful in the identification of human trophoblast membrane components that might influence immunological events
DISTRIBUTION AND ROLE ON FERTILIZATION OF PC~CINE ZON~ PELLUCIDA ANTIGENS ANALYZED BY MONOCLONAL ANTIBODIES S.Isojima, K.Koyama, A.Hasegawa, Y.Tsunoda* and A.Hanada* Department of Obstetrics and Gynecology, Hyogo Medical College, Nishinomiya, 663 and *National Institute of Animal Industry, Tsukuba, 305, Japan Five hybridomas which produce monoclonal antibodies (Moabs) to porcine zona pellucida (Z.P.) were established. The immunoglobulin classes were IgG2a from hybridoma BIIC8 and IgM from the other four. Under immunofluorescent staining, the Moab from BIIC8 stained only porcine oocytes, the Moab from GIOG5 stained oocytes of pigs and hamsters but Moabs from C6HI, D3H4, GIOF9 stained oocytes of pigs, humans, hamsters, rats and mice. Moabs from BIIC8 and GIOF9 strongly blocked boar sperm binding to porcine Z.P. but other Moabs only did slightly. However no Moab could block the sperm penetration on in vitro fertilization (IVF) of hamster oocytes. When a second antibody was applied after each Moab, only Moab from GIOG5 impaired IVF of Hamster oocytes. Two hybridomas producing Moabs to solubilized porcine Z.P. protein which reacted only to Z.P.s of pigs and humans were also selectively established. The immunoglobulin classes from both hybridomas (3A4-2GI, IDS-2B7) were IgGl. These Moabs stained specifically Z.P. but no other tissues of pigs by immunofluorescent staining. When human oocytes were pretreated with each Moab or a combination of two Moabs, no inhibitory effects on the sperm binding to oocytes were observed, whereas the pretreatment of human oocytes with conventional mouse antiserum to porcine Z.P. blocked completely the sperm binding to oocytes. When rabbit antiserum to mouse y-globulin was added as a second antibody to the human oocy~ tes pretreated with each Moab, the spex~ binding to oocytes was completed impaired with formation of a light scattering immunoprecipitin line on the surface of Z.P.
IMMUNOHISTOLOGICAL AND SEROLOGICAL APPLICATIONS OF MONOCLONAL ANTIBODIES TO HUMAN TROPHOBLAST MEMBRANE ANTIGENS IN PREGNANCY AND MALIGNANCY P.M. JOHNSON Dept. of Irmnunology, University of Liverpool, P.O. Box 147, Liverpool. L69 3BX, England. Monoclonal antibodies (mAbs) have proven useful in the identification of human trophoblast membrane components that might influence immunological events