Parainfluenza virus infection of cultured airway epithelial cells

Abstract

Studies of the cellular effects of respiratory viruses have generally used cultures of non-airway (particularly renal) epithelial cells. This requires the assumption that, despite the marked differences between renal epithelium and airway epithelium, the virus-host cell interactions in cultures of renal epithelium will be relevant to those in airway epithelium. To study viral infection of airway epithelial cells, we removed the epithelial cells from ferret tracheas using 0.1% pronase solution, and plated them at a density of 5 × 10 5 cells/cm 2 in collagen-coated plastic tissue culture wells. Cultures grew to confluence after 5–7 days. Viral inocula, consisting of supernatants from parainfluenza type 1-infected rhesus monkey kidney cell monolayers, were added to the culture medium in a concentration 10 3 times that sufficient to produce infection in 50% of rhesus monkey kidney monolayers (TCID 50). Cytopathic changes, consisting of cellular elongation and detachment, became apparent after 3–6 days, at which time the medium contained 5 × 10 8 TCID 50/ml. The monolayer appeared to be uniformly infected as revealed by adsorption of guinea pig erythrocytes. Specific immunofluorescence revealed uniformly positive staining for parainfluenza type 1 antigens. The ability to infect pure cultures of airway epithelial cells with viruses will allow us to examine the effects of these viruses on epithelial cell function, and to study virus-host cell interactions in cell cultures derived from the natural host cell.