Helicobacter pylori disrupts NADPH oxidase targeting in human neutrophils to induce extracellular superoxide release.
Helicobacter pylori (Hp) infection triggers a chronic influx of polymorphonuclear leukocyte neutrophils (PMNs) into the gastric mucosa. Although Hp reside in a neutrophil-rich environment, how these organisms evade phagocytic killing is largely unexplored. We now show that live Hp (strains 11637, 60190, DT61A, and 11916) are readily ingested by PMNs and induce a rapid and strong respiratory burst that is comparable to PMA. Relative to other particulate stimuli, Hp are more potent activators of PMNs than opsonized zymosan, Staphylococcus aureus, or Salmonella. Strikingly, biochemical and microscopic analyses demonstrate that Hp disrupt NADPH oxidase targeting such that superoxide anions are released into the extracellular milieu and do not accumulate inside Hp phagosomes. Specifically, nascent Hp phagosomes acquire flavocytochrome b558 but do not efficiently recruit or retain p47phox or p67phox. Superoxide release peaks at 16 min coincident with the appearance of assembled oxidase complexes in patches at the cell surface. Oxidant release is regulated by formalin-resistant and heat-sensitive bacterial surface factors distinct from urease and Hp(2-20). Following opsonization with fresh serum, Hp triggers a modest respiratory burst that is confined to the phagosome, and ingested bacteria are eliminated. We conclude that disruption of NADPH oxidase targeting allows unopsonized Hp to escape phagocytic killing, and our findings support the hypothesis that bacteria and PMNs act in concert to damage the gastric mucosa.
Allen
Lee-Ann H
LA
Department of Medicine and Inflammation Program, University of Iowa, Coralville, IA 52241, USA. lee-ann-allen@uiowa.edu
Beecher
Benjamin R
BR
Lynch
Jeffrey T
JT
Rohner
Olga V
OV
Wittine
Lara M
LM
D000328:Adult; D003573:Cytochrome b Group / Q000378:metabolism; D016480:Helicobacter pylori / Q000276:immunology / Q000472:pathogenicity*; D006801:Humans; D066298:In Vitro Techniques; D019255:NADPH Oxidases / Q000378:metabolism*; D009504:Neutrophils / Q000276:immunology* / Q000378:metabolism* / Q000382:microbiology; D009895:Opsonin Proteins / Q000008:administration & dosage; D010587:Phagocytosis; D010588:Phagosomes / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D016897:Respiratory Burst; D013481:Superoxides / Q000378:metabolism*
D003573:Cytochrome b Group; D009895:Opsonin Proteins; D010750:Phosphoproteins; C079789:neutrophil cytosol factor 67K; D013481:Superoxides; C036888:cytochrome b558; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1