Newborn mouse spleen, whose cells strongly suppress the in vitro humoral response of adult spleen cells, is essentially a hematopoietic organ. It contains a large percentage of proliferating cells, among which about 50% are erythroblasts (identified by their spectrin content) and about 15% are cells of the myelocytic and monocytic lineage. Lymphoid cells are a minority, with about 20% B and only 1-2% T lymphocytes. After a 4 days, a culture of newborn spleen cells contains 5-10 times more macrophages than that of an adult spleen. Most of these macrophage precursors from the newborn spleen are proliferating cells, partially glass- or plastic-adherent, which differentiate in culture into activated macrophages producing large amounts of plasminogen activator. It is this macrophage excess which is responsible for the immunosuppressive effect of newborn spleen cells in culture, as indicated by (a) the effect of silica particles added to the cultures, which both relieve the suppression and prevent the accumulation of macrophages and (b) the suppression of the humoral response of adult spleen cells when they are cultured on the adherent cells from a newborn but not from an adult spleen. The suppressive effect of macrophages seems to result, at least in part, from the production of prostaglandin, since it can be relieved by indomethacin or aspirin. Suppression is not related to arginine depletion of the medium or to production of an excess of plasminogen activator. T lymphocytes from newborn spleen or lymph nodes have no suppressive capability.