Interleukin-18 (IL-18) is a costimulatory factor for interferonγ (IFNγ) production. Processing of pro–IL-18 by IL-1β–converting enzyme (ICE) leads to the release of bioactive IL-18. Compared with wild-type (WT) mice, splenocytes from ICE-deficient mice produced low IFNγ after lipopolysaccharide (LPS) or zymosan (50% and 80% reduction). In contrast, IFNγ production was unimpaired in ICE-deficient mice using Concanavalin A (Con A). Comparable results were obtained when endogenous IL-18 was blocked with a neutralizing antibody. LPS-induced IFNγ was also reduced by an ICE inhibitor. Exogenous IL-18 augmented zymosan-induced IFNγ production in WT mice. In ICE-deficient cells, IFNγ production was only partially restored by IL-18. The reduced levels of IFNγ in ICE-deficient mice were not due to a lack of IL-12, because zymosan induced IL-12 equally in WT and in ICE-deficient mice. IFNγ is an important regulator of cell proliferation. In accordance, splenocytes from ICE-deficient mice proliferated more when stimulated with LPS, but not with Con A. Furthermore, in ovalbumin-sensitized ICE-deficient mice, proliferation of lymph node cells in response to the specific antigen was not altered. Exogenous IFNγ inhibited, whereas blockade of endogenous IFNγ or IL-18 increased, LPS induced splenocyte proliferation both in WT and in ICE-deficient mice. Our results show that IL-18 is an IL-12–independent regulator of IFNγ production and of cell proliferation induced by microbial stimuli. However, ICE-dependent processing of IL-18 is not needed for response to mitogens or antigens.