UCSF

While human or animal models are often considered the gold standard experimental system for defining virulence factors, cell culture-based infection models have proven useful for identifying important virulence factors and for examining the interactions between pathogens and the epithelial barrier. The first step in infections for most mucosal pathogens involves binding (adhesion) to the epithelial cells that line the mucosa. Successful pathogens can then penetrate the barrier by (1) inducing their uptake (i.e., "entry" or "invasion") into epithelial cells, (2) crossing the barrier by inducing epithelial cell death, and/or (3) penetrating between cells. This chapter describes growth conditions to form polarized cultures, either two-dimensional monolayers or three-dimensional cysts, of various immortalized epithelial cell lines. It describes assays to measure key early interactions between P. aeruginosa and host cells, including binding, invasion, and cytotoxicity. Many virulence factors defined by these criteria have been shown to be important for pathogenesis of P. aeruginosa infections in animals or humans. These methods are also applicable to other pathogens.