Attachment of influenza A virus to ferret tracheal epithelium at different maturational stages.

Influenza virus attaches primarily to ciliated cells in mature airways epithelium. This process is mediated by a viral envelope glycoprotein (hemagglutinin) that binds to sialic acid-containing receptors in the apical membrane of host cells. The purpose of this study was to determine the cellular distribution of these receptors as a function of tracheal epithelial maturation in the ferret, which is susceptible to influenza virus infection at all ages and undergoes postnatal ciliation. To assay for virus attachment, tracheal strips from ferrets at ages 0, 7, 14, and 28 d were incubated at 4 degrees C for 1 h with a concentrated suspension of influenza A virus. Transmission electron microscopy demonstrated virus attachment to the apical surface of 77 to 87% of ciliated cells, but only to 1 to 9% of nonciliated surface epithelial cells at all ages, including the newborn, which has few ciliated cells (less than 10% of total cells). Virions also attached to most of the preciliated cells identified. Pretreatment of tracheal strips with neuraminidase virtually eliminated viral attachment. These findings demonstrate preferential influenza virus binding to sialylated receptors on ciliated cells and their immediate precursors. The sparsity of ciliated cells with no evidence for increased influenza virus binding per cell in newborn ferret tracheas suggests that the previously demonstrated high risk of death from influenza infection in newborn ferrets is due to factors other than increased susceptibility to virus attachment. Influenza virus receptors appear to be selective membrane markers for ciliated cells and may be particularly useful for the identification of preciliated cells.