In the mare, early pregnancy loss is common, but involvement of the maternal immune system in the pathogenesis of this condition has not been investigated in detail so far. In the present study, we assessed effects of exposure of the endometrium to semen or seminal plasma in oestrous mares on the response of regulatory T lymphocytes (Tregs) in the peripheral circulation as well as in the endometrium. Raw semen, seminal plasma or PBS (control) were introduced into the uterus of oestrous mares (n = 12). Blood was collected immediately before insemination or PBS infusion (time 0), and 12, 24 and 48 h thereafter. Endometrial biopsies were collected at 24 h. In peripheral blood, Treg (CD4+Foxp3+) cells were determined by flow cytometry. In endometrial biopsies, Tregs were assessed as cells staining positive for Foxp3 by immunohistochemistry. The percentage of Tregs in blood decreased (p < 0.05) at 12 h after exposure to seminal plasma, tended to decrease in response to raw semen (p = 0.095) but not to PBS. Leukocyte and PMN counts were not affected. In the endometrium, numbers of Foxp3 positive cells at 24 h after insemination or PBS infusion were not changed by treatment. Results of the present study provide only little evidence that maternal tolerance of pregnancy in the horse is modulated already by exposure of the oestrous endometrium to seminal plasma at mating.