Pseudomonas aeruginosa cystic fibrosis isolates induce rapid, type III secretion-dependent, but ExoU-independent, oncosis of macrophages and polymorphonuclear neutrophils.
Pseudomonas aeruginosa, an opportunistic pathogen responsible most notably for severe infections in cystic fibrosis (CF) patients, utilizes the type III secretion system for eukaryotic cell intoxication. The CF clinical isolate CHA shows toxicity towards human polymorphonuclear neutrophils (PMNs) which is dependent on the type III secretion system but independent of the cytotoxin ExoU. In the present study, the cytotoxicity of this strain toward human and murine macrophages was demonstrated. In low-multiplicity infections (multiplicity of infection, 10), approximately 40% of the cells die within 60 min. Analysis of CHA-infected cells by transmission electron microscopy, DNA fragmentation assay, and Hoechst staining revealed the hallmarks of oncosis: cellular and nuclear swelling, disintegration of the plasma membrane, and absence of DNA fragmentation. A panel of 29 P. aeruginosa CF isolates was screened for type III system genotype, protein secretion profile, and cytotoxicity toward PMNs and macrophages. This study showed that six CF isolates were able to induce rapid ExoU-independent oncosis on phagocyte cells.
Dacheux
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D
Département de Biologie Moléculaire et Structurale, BBSI, UMR-314 CNRS, CEA-Grenoble, France.
Toussaint
B
B
Richard
M
M
Brochier
G
G
Croize
J
J
Attree
I
I
D036002:ADP Ribose Transferases / Q000378:metabolism*; D000818:Animals; D017209:Apoptosis; D001402:B-Lymphocytes / Q000166:cytology; D001426:Bacterial Proteins / Q000378:metabolism*; D001427:Bacterial Toxins / Q000378:metabolism*; D001562:Benzimidazoles; D002460:Cell Line; D003550:Cystic Fibrosis / Q000382:microbiology*; D003603:Cytotoxins / Q000378:metabolism; D053938:DNA Fragmentation; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005456:Fluorescent Dyes; D005964:Glucosyltransferases / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008264:Macrophages / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D008854:Microscopy, Electron; D009504:Neutrophils / Q000166:cytology* / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000302:isolation & purification / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods; D013997:Time Factors; D015534:Trans-Activators / Q000378:metabolism*
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D001562:Benzimidazoles; D003603:Cytotoxins; D004268:DNA-Binding Proteins; C095092:ExsA protein, bacteria; D005456:Fluorescent Dyes; D015534:Trans-Activators; C111749:pseudomonas exoprotein A protein, Pseudomonas aeruginosa; C111504:ExoY protein, bacteria; D005964:Glucosyltransferases; D036002:ADP Ribose Transferases; C017807:bisbenzimide ethoxide trihydrochloride