R-025. Effect of peritoneal fluid from fertile women and from infertile women with endometriosis or unexplained infertility on the acrosome reaction of capacitated spermatozoa

or an influence on sperm motility and the acrosome reaction was observed.Materials and methods: Spermatozoa from ejaculates of healthy donors with normal sperm parameters were washed twice in IVF buffer and prepared using the swim-up procedure.Isolation and purification of the membrane proteins were achieved by solubilization in demembranated medium, stripping of the membrane with a homogenizer, sonication and several steps of ultracentrifugation.The highly enriched human sperm membrane proteins were separated using a two-dimensional electrophoresis technique.Isoelectric focusing, the first dimension, was performed on precast immobiline dry strip gels at pH 4-7.The second dimension of electrophoresis was carried out on 12% SDS-polyacrylamide gels.The proteins in acrylamide gels were stained with Coomassie Blue R® or silver.Proteins were electrotransferred onto polyvinylidine difluoride sheets and stained with Coomassie blue R® or exposed to seminal plasma.Seminal plasma samples from patients containing antisperm antibodies and control samples underwent immunoblotting to identify the antigenic proteins.Results: Some antigens were detected with the seminal plasma antibodies.The molecular weights and isoelectric points were determined.Some antigens were common in many patients, while other antigens were detected only in individual cases. Conclusion:The result of this study will help to classify the phenomenon of immunological infertility.