Incidence of anti-zona pellucida and anti-sperm antibodies among infertile Jordanian women and its relation to mycoplasmas

Anti-zona-pellucida autoantibodies [AZP-Ab] and anti-sperm isoantibodies [ASA] were assessed in the cervical secretions from 73 infertile Jordanian women and 41 fertile control women using latex agglutination. Significantly more women with infertility had AZP-Ab and ASA [16.4% and 8.2% respectively] compared with fertile women [9.4% and 0%], with no relation to the etiology of infertility. Using polymerase chain reaction Mycoplasma hominis and Ureaplasma urealyticum were detected in cervical secretions of 19.2% and 13.7% of infertile women, and the presence of mycoplasma was significantly correlated with the presence of AZP-Ab and ASA

Antígenos de zona pelúcida en el ovocio humano: su importancia en las estrategias anticonceptivas / Zona pellucida antigens of human oocyte: its relevance in contraceptive strategies

La zona pelúcida (ZP) es una capa de glicoproteínas que rodea al ovocito de los mamíferos. Esta cubierta está formada por tre familias de glicoproteínas denominadas ZP1, ZP2 y ZP3, que difieren en sus propiedades inmunológicas y funcionales debido a modificaciones postraduccionales. Estudios llevados a cabo en el ratón sugieren que la función de estas proteínas se encuentra relacionada con el reconocimiento del espermatozoide por la ZP, confiriéndole muy probablemente la connotación de receptores. Esta observación ha permitido que varios laboratorios hayan iniciado la producción y obtención de las proteínas de la ZP, permitiendo la exploración de su papel en procesos fisiológicos y clínicos, y ha abierto la posibilidad de utilizarlas en el desarrollo de un método inmunológico anticonceptivo. En la actualidad la posibilidad de obtener anticuerpos específicos contra los constituyentes proteínicos de la ZP representa una estrategia novedosa para el control de la fertilidad en el humano.

Characterization of antigenic determinants on porcine zona pellucida-3 beta(ZP3 beta) glycoprotein by monoclonal antibodies.

Out of 11 monoclonal antibodies(MAbs) developed against porcine zona pellucida-3 beta(ZP3 beta) glycoprotein, 6 (MA-451, -454, -455, -462, -467 and -470) reacted with ZP2 beta, deglycosylated ZP3 beta(DGZP3 beta), and reduced and carboxyamidomethylated ZP3 beta(RCMZP3 beta) in ELISA and Western blot suggesting that these monoclonals recognise a linear protein epitope. Five MAbs(MA-452, -456, -459, -474 and -476) showed reduced binding with DGZP3 beta and RCMZP3 beta as compared to ZP3 beta in ELISA suggesting that these may recognize either carbohydrate moities or conformation dependent epitopes. Moreover, these MAbs failed to recognize these antigens on Western blot. Based on competitive inhibition studies in ELISA for the ability of MAbs to inhibit the binding of biotinylated ZP3 beta to solid phase antibody, 6 distinct domains were discernible. Domain V recognised by MA-467 partially overlaps with the domain corresponding to MA-454 and MA-455. All 11 MAbs reacted with zona pellucida of intact oocytes as revealed by indirect immunofluorescence but only 3 antibodies (MA-454, -455 and -467) delayed the zona lysis by trypsin. In immunoblots MA-467 recognised 18kDa fragment of RCMZP3 beta digested with endoproteinase lys-C and 37 and 30kDa bands of elastase digest of ZP3 beta. The results will help in delineation of functionally relevant domains and design of contraceptive vaccine based on zona pellucida aiming for pre-fertilization block.

Generation and characterization of monoclonal antibodies to porcine zona pellucida-3 alpha glycoprotein.

All 10 monoclonal antibodies generated reacted with deglycosylated ZP3 alpha both in ELISA and Western blots, thereby suggesting that these do not recognize carbohydrate determinants. Moreover, 7 antibodies (MAs -402, -403, -405, -412, -420, -421 and -423) recognized reduced and carboxyamidomethylated ZP3 alpha in Western blot suggesting that these antibodies recognized linear epitopes. The epitopes recognized by MAs -410, -413 and -425 were sequential or conformational, stabilized by disulphide bonds. Three antibodies namely, MA -405, -420 and -421 inhibited in vitro, zona lysis by trypsin.