Induction of necrosis in human neutrophils by Shigella flexneri requires type III secretion, IpaB and IpaC invasins, and actin polymerization.
Infection by Shigella flexneri is characterized by infiltration of neutrophils in the intestinal mucosa and by a strong inflammatory reaction. Although neutrophils are constitutively programmed to die by apoptosis, we show that isolated human neutrophils undergo necrosis 2 h after infection with virulent S. flexneri strain M90T but not with the virulence plasmid-cured strain BS176. This was demonstrated by the release of azurophil granule proteins concomitant with the release of lactate dehydrogenase (LDH), disruption of the plasma membrane, and absence of DNA fragmentation. Mutants with the mxiD1 gene, coding for an essential component of the secretion type III machinery, or the genes coding for IpaB or IpaC invasins deleted were not cytotoxic. Neutrophil necrosis occurred independently of the bacterial ability to leave phagosomes, and it involved actin polymerization, as the addition of cytochalasin D after phagocytosis of Shigella inhibited the release of LDH. In conclusion, Shigella kills neutrophils by necrosis, a process characterized by the release of tissue-injurious granular proteins. This probably contributes to disruption of the epithelial barrier, leading to the dysentery observed in shigellosis and allowing Shigella to enter its host cells.
François
M
M
Institut de Pharmacologie et de Biologie Structurale, CNRS UPR 9062, Toulouse, France.
Le Cabec
V
V
Dupont
M A
MA
Sansonetti
P J
PJ
Maridonneau-Parini
I
I
D000199:Actins / Q000378:metabolism*; D000942:Antigens, Bacterial / Q000502:physiology*; D017209:Apoptosis; D001426:Bacterial Proteins / Q000502:physiology*; D005089:Exocytosis; D006801:Humans; D008247:Lysosomes / Q000502:physiology; D008561:Membrane Fusion; D009336:Necrosis; D009504:Neutrophils / Q000473:pathology*; D010587:Phagocytosis; D011108:Polymers / Q000378:metabolism; D012763:Shigella flexneri / Q000276:immunology*; D014617:Vacuoles / Q000382:microbiology
D000199:Actins; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D011108:Polymers; C087447:ipaB protein, Shigella; C092342:IpaC protein, Shigella