title: Afa/Dr-expressing, diffusely adhering Escherichia coli strain C1845 triggers F1845 fimbria-dependent phosphatidylserine externalization on neutrophil-like differentiated PLB-985 cells through an apoptosis-independent mechanism.
abstract: The enterovirulent Escherichia coli strains potentially involved in inflammatory bowel diseases include diffusely adherent strains expressing Afa/Dr fimbriae (Afa/Dr DAEC). We have previously observed type 1 pilus-mediated interleukin-8 (IL-8) hyperproduction in infected neutrophils. As pathogen induction of host cell death programs and clearance of apoptotic infected cells are crucial for innate immune system homeostasis and host integrity, we examined modulation of neutrophil cell death by Afa/Dr DAEC. Using the human PLB-985 cell line differentiated into fully mature neutrophils, we found that the wild-type enterovirulent E. coli strain C1845 and the recombinant strain DH5alpha/pF1845 (expressing the fimbrial adhesin F1845) similarly induced time-dependent phosphatidylserine (PS) externalization, suggesting a major specific role of this virulence factor. Using small interfering RNA (siRNA) decay-accelerating factor (DAF)-transfected PLB-985 cells, we then showed that this PS externalization was triggered in part by glycosylphosphatidylinositol (GPI)-anchored DAF receptor engagement (leading to tyrosine kinase and protein kinase C activation) and that it required cytoskeleton and lipid raft architectural integrity. PS externalization under these conditions was not dependent on caspases, mitochondria, lysosomes, or reactive oxygen or nitrogen species. F1845-mediated PS externalization was sufficient to enable macrophage engulfment of infected differentiated PLB-985 cells. These findings provide new insights into the neutrophil response to Afa/Dr DAEC infection and highlight a new role for F1845 fimbriae. Interestingly, although apoptosis pathways were not engaged, C1845-infected PLB-985 cells displayed enhanced removal by macrophages, a process that may participate in the resolution of Afa/Dr DAEC infection and related inflammation.
authors.affiliation: Inserm, UMR756 Signalisation et Physiopathologie des Cellules Epithéliales, and Université Paris-Sud XI, Faculté de Pharmacie, Châtenay-Malabry, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Sémiramoth
authors.forename: Aude
authors.initials: A
authors.lastname: Gleizes
authors.forename: Isabelle
authors.initials: I
authors.lastname: Turbica
authors.forename: Catherine
authors.initials: C
authors.lastname: Sandré
authors.forename: Viviana
authors.initials: V
authors.lastname: Marin-Esteban
authors.forename: Roseline
authors.initials: R
authors.lastname: Gorges
authors.forename: Alain
authors.initials: A
authors.lastname: Servin
authors.forename: Sylvie
authors.initials: S
authors.lastname: Chollet-Martin
mesh_terms: D018830:Adhesins, Escherichia coli / Q000502:physiology; D017209:Apoptosis / Q000502:physiology; D001422:Bacterial Adhesion / Q000502:physiology; D015153:Blotting, Western; D045744:Cell Line, Tumor; D054308:Enteropathogenic Escherichia coli / Q000502:physiology*; D004927:Escherichia coli Infections / Q000382:microbiology; D029968:Escherichia coli Proteins / Q000502:physiology*; D036781:Fimbriae Proteins / Q000502:physiology; D010861:Fimbriae, Bacterial / Q000502:physiology*; D006098:Granulocytes / Q000382:microbiology / Q000502:physiology; D006801:Humans; D008247:Lysosomes / Q000382:microbiology / Q000502:physiology; D008264:Macrophages / Q000382:microbiology / Q000502:physiology; D009504:Neutrophils / Q000382:microbiology / Q000502:physiology; D010587:Phagocytosis / Q000502:physiology; D014157:Transcription Factors / Q000502:physiology*
chemical_list: D018830:Adhesins, Escherichia coli; C080953:DaaA protein, E coli; D029968:Escherichia coli Proteins; D014157:Transcription Factors; D036781:Fimbriae Proteins
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