ABSTRACT: Pregnant and progesterone-treated, ovariectomized ewes accumulate secretory products in the uterus that are immunosuppressive in mitogen-stimulated and mixed lymphocyte sheep cell cultures. In this study, uterine secretions from pregnant (Preg-UTM) and progesterone-treated, ovariectomized (P-UTM) ewes were equally effective in suppressing 3H-Tdr incorporation in mouse spleen cells stimulated with PHA. P-UTM inhibited PHA-stim-ulated, purified T-cells and separated L3T4+ and Lyt2+ T-cell subpopulations more than Preg-UTM, however. Both fluids were slightly inhibitory to conA-stimulated mouse spleen cells, enriched T-cells, and Lyt2+ T-cells but neither inhibited L3T4+ T-cells. For LPS-stimulated cells, P-UTM caused more suppression than Preg-UTM of enriched B-cells; however, suppression was similar for the two fluids on unseparated splenic cells. In antigen-stimulated mouse spleen cell cultures, both fluids inhibited antibody-forming cell responses to sheep erythrocytes, a thymus-dependent antigen, but neither suppressed antibody-forming cell responses to TNP-Ficoll, a thymus-independent antigen. These data indicate that uterine secretions in the ewe produced under the influence of progesterone or pregnancy contain immunoregulatory molecule(s) which modulate the activity of both homologous sheep and unrelated mouse lymphocytes. These studies establish the mouse as a useful in vivo model for studying the biological actions of these molecules.