The linkage between in utero exposure to diethylstilbestrol (DES) and the manifestation of a variety of reproductive disorders and possibly immune alterations in adults (i.e., human and mice) is suggestive of a fetal basis of adult disease. While the long-term adverse consequences of prenatal DES-exposure on reproductive disorders are well known, there is a paucity of data with regard to immune outcome. We hypothesize that prenatal DES-exposure “imprints” the immune system, altering the response to subsequent exposure to DES in adult mice. In this pilot study, C57BL/6 mice were prenatally exposed to DES or vehicle only (oil) and then exposed to DES at 1 year of age. Potential alterations in the spleen were then examined. Female DES-exposed mice (DES prenatal /DES adult ) or female DES had higher serum levels of interferon-gamma (IFNγ) in response to administration of an IFNγ -inducer (soluble proteins-derived from Toxoplasma gondii), compared to female controls, which received oil during prenatal life (Oil prenatal /DES adult ). Splenic lymphocytes from female DES prenatal /DES adult mice, when activated with Concanavalin A (ConA), also secreted higher levels of IFNγ compared to female controls (Oil prenatal /DES adult ) when examined at 14-months of age. This increase in IFNγ in prenatal DES-exposed mice is not due to enhanced numbers of splenocytes or increased relative percentages of CD4 + or CD8 + cells. ConA-activated T-cells from female DES prenatal /DES adult had increased expression of the co-stimulatory molecule, CD28. These above immune changes were not evident in the males prenatally exposed to DES. Prenatal DES exposure also did not induce autoimmunity in non-autoimmune C57BL/6 mice. Overall, results from these prefatory studies suggest that prenatal DES exposure may have long-term immune alterations, which become evident following a secondary exposure to DES in adult life. [ABSTRACT FROM AUTHOR]