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Different mechanisms have been proposed, including virus-induced damage to respiratory cells predisposing to opportunistic bacterial infection or up-regulation of bacterial adhesion molecules by viral infection [13�C15]. The aim of the present study was to analyse whether bacterial exposure can influence susceptibility to HMPV infection. First, we demonstrated for 57 healthy children that they were immune-competent; indeed, all children had seroconverted to MV at the age of 24?months, approximately half a year after MMR vaccination. Subsequently, we screened the colonization state of these 57 children for four common respiratory bacterial species (H.?influenzae, M.?catarrhalis, S.?aureus and S.?pneumoniae) in relation to seroconversion to HMPV during the first 2?years of life. Whereas no relationship was detected between Selleckchem Ribociclib exposure to H.?influenzae, M.?catarrhalis or S.?aureus and HMPV seroconversion, S.?pneumoniae exposure was significantly associated with increased seroconversion levels to HMPV. These increased HMPV seroconversion levels could be due to increased susceptibility to HMPV infection, increased viral replication or virus spread, or enhanced immune responses to infection. Differences in bacterial exposure and HMPV seroconversion could also be related to a common external factor (e.g. attendance at a day-care centre). However, in that case we would expect that the carrier state of H.?influenzae, M.?catarrhalis, S.?aureus and S.?pneumoniae would also be affected. On the basis of the serology data, we concluded that either Otenabant HMPV infection leads to more frequent S.?pneumoniae carriage or exposure to S.?pneumoniae increases the susceptibility to HMPV infections. To test our hypothesis that bacterial exposure could increase the susceptibility to HMPV infection, we used wd-NHBE cells cultured on the air-liquid interface. The cells clearly showed beating cilia and mucus production, and may therefore be considered the best possible in vitro mimic of the target cells of HMPV infection in vivo. Surprisingly, apical infection of wd-NHBE cells with HMPV-EGFP at an estimated MOI of 10 resulted in a low frequency MI-773 cost (