This project is to provide a foundation for the prevention of endometriosis-associated reproductive failure through the identification of etiologic factors, new diagnostic assays, and best predictors of successful therapy. Our objectives are to further define immunological mechanisms that are involved in the pathogenesis of endometriosis-associated reproductive failure, and to develop new diagnostic approaches based on these mechanisms for detection and treatment of this disease. Immunohistologic and Northern/slot-blot analyses will be performed to detect white blood cell populations, cytokine-positive cells and cytokine mRNA levels in biopsies of ectopic (endometriosis) and eutopic (intrauterine) endometrium. Peritoneal fluid cells will be immunohistologically characterized and quantified. Immunoassays will be used to measure selected cytokine and C3 levels in peritoneal fluid and peripheral serum to identify potential new markers for further characterization and diagnosis of endometriosis-associated reproductive failure. Peritoneal fluid containing elevated levels of cytokines of C3 components will be tested for effects on in vitro models of reproductive events including: human sperm motility; hamster egg-human sperm interaction; and murine embryo viability, development and implantation events. Factors in peritoneal fluid inhibiting these assays will be characterized using immunoaffinity chromatography and other biochemical methods. Immunological variables will be correlated with epidemiological variables to determine the role of these factors in the development of endometriosis-associated reproductive failure by prospectively following patients to determine the occurrence and outcome of subsequent pregnancy.
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