ULTRASTRUCTURAL ANALYSIS OF PRIMARY HUMAN URETHRAL EPITHELIAL-CELL CULTURES INFECTED WITH NEISSERIA-GONORRHOEAE

In men with gonococcal urethritis, the urethral epithelial cell is a s ite of infection. To study the pathogenesis of gonorrhea in this fell type, we have developed a method to culture primary human urethral epi thelial cells obtained at the time of urologic surgery. Fluorescent an alysis demonstrated that 100% of the cells stained for keratin, Micros copic analyses indicated that these epithelial cells arrayed in a patt ern similar to that seen in urethral epithelium. Using immunoelectron and confocal microscopy, we compared the infection process seen in pri mary cells with events occurring during natural infection of the same cell type in men,vith gonococcal urethritis. Immunoelectron microscopy studies of cells infected with Neisseria gonorrhoeae 1291 Opa(+) P+ s howed adherence of organisms to the epithelial cell membrane, pedestal formation with evidence of intimate association between the gonococca l and the epithelial cell membranes, and intracellular gonococci prese nt in vacuoles, Confocal studies of primary urethral epithelial cells showed actin polymerization upon infection, Polyclonal antibodies to t he asialoglycoprotein receptor (ASGP-R) demonstrated the presence of t his receptor on infected cells in the primary urethral cell culture. I n situ hybridization using a fluorescent-labeled probe specific to the ASGP-R mRNA demonstrated this message in uninfected and infected cell s, These features were identical to those seen in urethral epithelial cells in exudates from males with gonorrhea, Infection of primary uret hral cells in culture mimics events seen in natural infection and will allow detailed molecular analysis of gonococcal pathogenesis in a hum an epithelial cell which is commonly infected.