Defensins are effector molecules of the innate immunity with a broad antimicrobial spectrum, including HIV. They also link innate and adaptive immunity, displaying chemotactic activity for monocytes, T cells, and dendritic cells (DCs). alpha-Defensins 1-3 are mainly produced by neutrophils, but their production by other leukocyte subsets has also been reported. Herein, we studied whether monocyte-derived DCs (MDDCs), which are regarded as a model for myeloid DCs, produce alpha-defensins 1-3. We found that immature MDDCs (imMDDCs) produce alpha-defensins 1-3 mRNA, but this production is undetectable or barely detectable following 48 h of maturation with the proinflammatory cytokine cocktail (IL-1beta+IL-6+TNF-alpha) or LPS. It is surprising that alpha-defensins 1-3 production was up-regulated when exposed to each one of the proinflammatory cytokines alone, especially IL-1beta. alpha-Defensins 1-3 produced by imMDDCs were mainly secreted peptides. Production and secretion of alpha-defensins 1-3 by imMDDCs can have biological relevance for the antigen processing of pathogens and can contribute to understanding differences in susceptibility to infections, an issue of special interest in the field of HIV infection.