Short-term stimulation of mouse spleen cells in vitro with interleukin (IL)-3 induces the secretion of the Th2 cytokines IL-4 and IL-6. Non-B/non-T cells were the target of this IL-3 effect. However, during long-term antigen-dependent culture, T cells are the major source of IL-4 and IL-6. The addition of IL-3 to such cultures also led to a significant increase in IL-4 and IL-6 production. This Th2 cytokine secretion was amplified by the addition of irradiated non-B/non-T cells at the initiation of culture, and was inhibited by anti-IL-4 antibodies. These findings suggest that IL-3 induces the rapid release of IL-4 and IL-6 by non-B/non-T cells, thereby creating an immune milieu conducive to the development of antigen-specific IL-4 and IL-6-secreting Th2 cells.