Purification of outer membrane vesicles from Pseudomonas aeruginosa and their activation of an IL-8 response.
Considerable lung injury results from the inflammatory response to Pseudomonas aeruginosa infections in patients with cystic fibrosis (CF). The P. aeruginosa laboratory strain PAO1, an environmental isolate, and isolates from CF patients were cultured in vitro and outer membrane vesicles from those cultures were quantitated, purified, and characterized. Vesicles were produced throughout the growth phases of the culture and vesicle yield was strain-independent. Strain-dependent differences in the protein composition of vesicles were quantitated and identified. The aminopeptidase PaAP (PA2939) was highly enriched in vesicles from CF isolates. Vesicles from all strains elicited IL-8 secretion by lung epithelial cells. These results suggest that P. aeruginosa colonizing the CF lung may produce vesicles with a particular composition and that the vesicles could contribute to inflammation.
Bauman
Susanne J
SJ
Duke University Medical Center, Department of Biochemistry, Box 3711, Research Drive, Durham, NC 27710, USA.
Kuehn
Meta J
MJ
D001425:Bacterial Outer Membrane Proteins / Q000032:analysis / Q000737:chemistry; D003550:Cystic Fibrosis / Q000382:microbiology*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004847:Epithelial Cells / Q000276:immunology / Q000378:metabolism; D006801:Humans; D016209:Interleukin-8 / Q000276:immunology / Q000378:metabolism*; D008168:Lung / Q000276:immunology / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods; D011552:Pseudomonas Infections / Q000382:microbiology*; D011550:Pseudomonas aeruginosa / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D022142:Secretory Vesicles / Q000737:chemistry* / Q000276:immunology* / Q000378:metabolism
D001425:Bacterial Outer Membrane Proteins; D016209:Interleukin-8