Various inflammatory cells and cytokines have been identified in otitis media with effusion (OME). The presence of neutrophils has been linked to interleukin-8, but no chemotactic factor has as yet been identified for monocytes. The chemokine RANTES (Regulated upon Activation, Normal T Expressed and Secreted) attracts and activates primarily monocytes and may contribute to the pathogenesis of middle ear inflammation. We investigated the presence of RANTES by: 1) ELISA measurement in 114 middle ear effusions from children suffering from OME, 2) immunohistochemical localisation in experimental OME rabbit middle ear mucosa, and 3) expression in cultured rabbit middle ear epithelium in response to proinflammatory stimuli. RANTES was detectable in 94 (82%) of 114 effusions with a median concentration of 79.7 pg/mg total protein content. The concentration of RANTES was positively correlated with the endotoxin content. Immunohistochemically, RANTES was localized to the epithelial layer in experimental OME. In vitro, RANTES was expressed in middle ear epithelium in response to proinflammatory stimuli (TNF-α) in a dose-dependent manner. The expression of RANTES may explain the recruitment of monocytes in OME, possibly as a result of TNF-α-mediated endotoxin stimulation.