Role of myeloperoxidase in the killing of Staphylococcus aureus by human neutrophils: studies with the myeloperoxidase inhibitor salicylhydroxamic acid.
We have used salicylhydroxamic acid (SHAM) to inhibit intraphagosomal myeloperoxidase activity in order to evaluate the role of this enzyme in the killing of Staphylococcus aureus by human neutrophils. 50 microM-SHAM reduced the luminol-dependent chemiluminescence response stimulated during phagocytosis of unopsonized latex beads and opsonized S. aureus by over 80% and 60%, respectively. When opsonized S. aureus were incubated with neutrophils, 45% were killed within 15 min incubation and 60% by 1 h. However, in neutrophil suspensions incubated with 50 microM-SHAM, only 13% were killed by 15 min whilst 71% still remained viable after 1 h. This inhibitor had no effect upon the number of bacteria phagocytosed or upon degranulation. In a cell-free system, 2.5 microM-H2O2 alone killed 55% of the bacteria, whereas in the presence of myeloperoxidase (i.e. 10 mU myeloperoxidase and 2.5 microM-H2O2) virtually all of the bacteria were killed: the addition of 50 microM-SHAM abolished this myeloperoxidase-enhanced killing but did not affect the H2O2-dependent killing. We therefore conclude that in normal neutrophils whilst H2O2 is required for killing of this pathogen, both myeloperoxidase-dependent and -independent pathways exist.
Humphreys
J M
JM
Department of Biochemistry, University of Liverpool, UK.
Davies
B
B
Hart
C A
CA
Edwards
S W
SW
D015550:Cell Degranulation; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D008163:Luminescent Measurements; D009504:Neutrophils / Q000201:enzymology* / Q000502:physiology; D009895:Opsonin Proteins / Q000276:immunology; D010100:Oxygen / Q000378:metabolism; D009195:Peroxidase / Q000037:antagonists & inhibitors / Q000502:physiology*; D010587:Phagocytosis; D012457:Salicylamides / Q000494:pharmacology*; D013211:Staphylococcus aureus
D009895:Opsonin Proteins; D012457:Salicylamides; C005703:salicylhydroxamic acid; D006861:Hydrogen Peroxide; D009195:Peroxidase; D010100:Oxygen