Release of host-derived membrane vesicles following pilus-mediated adhesion of Neisseria gonorrhoeae

Abstract

Following attachment of Neisseria gonorrhoeae to human epithelial cell lines, the cellular pilus receptor CD46 is shed from the cell and accumulates in the media. In this report, we assess Neisseria-induced alterations in CD46 surface distribution and characterize this complement regulatory protein following its release from the infected cell. Within 3 h of attachment of gonococci to human epithelial cell lines, CD46 is enriched beneath sites of microcolony adhesion. By 6 h post infection, differential ultracentrifugation of culture media from ME-180 monolayers resulted in sedimentation of structurally and functionally intact CD46. Electron microscopy of these 100 000 g pellets revealed 30-200 nm vesicles. These vesicles likely originated from the host cell as they contained additional host cell surface proteins including CD55 and the epidermal growth factor receptor. Further, these vesicles were visualized by quick-freeze, deep-etch electron microscopy in association with the surface of infected ME-180 cells and with pili of adherent gonococci. Like CD46 shedding, CD46 redistribution and vesicle release were insensitive to colchicine and cytochalasin-D but dependent on expression of the pilus retraction protein PilT. This vesiculation may represent a host cell defence response in which surface proteins that are commonly exploited by pathogens, such as CD46, are removed from the cell. © 2005 Blackwell Publishing Ltd.