Autophagy is a key regulator of cellular homeostasis that can be activated by pathogen-associated molecules and recently has been shown to influence IL-1beta secretion by macrophages. However, the mechanisms behind this are unclear. Here, we describe a novel role for autophagy in regulating the production of IL-1beta in antigen-presenting cells. After treatment of macrophages with Toll-like receptor ligands, pro-IL-1beta was specifically sequestered into autophagosomes, whereas further activation of autophagy with rapamycin induced the degradation of pro-IL-1beta and blocked secretion of the mature cytokine. Inhibition of autophagy promoted the processing and secretion of IL-1beta by antigen-presenting cells in an NLRP3- and TRIF-dependent manner. This effect was reduced by inhibition of reactive oxygen species but was independent of NOX2. Induction of autophagy in mice in vivo with rapamycin reduced serum levels of IL-1beta in response to challenge with LPS. These data demonstrate that autophagy controls the production of IL-1beta through at least two separate mechanisms: by targeting pro-IL-1beta for lysosomal degradation and by regulating activation of the NLRP3 inflammasome.