title: IpaB mediates macrophage apoptosis induced by Shigella flexneri.
abstract: Shigella flexneri kills macrophages through apoptosis, involving the induction of host cell DNA fragmentation and characteristic morphological changes. Shigella can only cause damage if it escapes from the phagolysosome into the cytoplasm. The S. flexneri cytotoxic genes have been localized to the ipa operon of shigella's virulence plasmid. ipaB, C and D deletion mutants are not invasive and therefore not cytotoxic. In order to distinguish genes involved in the escape from the phagolysosome as distinct from cytotoxicity, we constructed Shigella strains that secrete low amounts of Escherichia coli haemolysin (hly(low)). These strains can escape into the cytoplasm of the macrophage even in the absence of the invasion plasmid as verified by electron microscopy and resistance to chloroquine. Macrophages were infected with different ipa mutants expressing hly(low). Both delta ipaC hly(low) and delta ipaD hly(low) were cytotoxic whilst delta ipaB hly(low) and a hly(low) strain cured of shigella's pathogenicity plasmid were not. Furthermore, both delta ipaC hly(low) and delta ipaD hly(low) killed through apoptosis as shown by both changes in ultrastructural morphology and fragmentation of the host cell DNA. These results demonstrate that ipaB is essential for S. flexneri to induce apoptosis in macrophages.
authors.affiliation: Unité de Pathogénie Microbienne Moléculaire, INSERM U199, Université de Paris Sud, Orsay, France.
authors.forename: A
authors.initials: A
authors.lastname: Zychlinsky
authors.forename: B
authors.initials: B
authors.lastname: Kenny
authors.forename: R
authors.initials: R
authors.lastname: Ménard
authors.forename: M C
authors.initials: MC
authors.lastname: Prévost
authors.forename: I B
authors.initials: IB
authors.lastname: Holland
authors.forename: P J
authors.initials: PJ
authors.lastname: Sansonetti
mesh_terms: D000818:Animals; D017209:Apoptosis; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D002738:Chloroquine / Q000494:pharmacology; D029968:Escherichia coli Proteins; D005839:Gentamicins / Q000494:pharmacology; D006460:Hemolysin Proteins / Q000235:genetics / Q000378:metabolism; D008247:Lysosomes / Q000382:microbiology; D008264:Macrophages / Q000382:microbiology*; D051379:Mice; D009876:Operon; D010587:Phagocytosis; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017384:Sequence Deletion; D012763:Shigella flexneri / Q000187:drug effects / Q000235:genetics / Q000472:pathogenicity*; D014774:Virulence / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D005839:Gentamicins; D006460:Hemolysin Proteins; C036507:Hlya protein, E coli; D011993:Recombinant Fusion Proteins; C087447:ipaB protein, Shigella; D002738:Chloroquine