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Enhancement of chemotactic factor-stimulated neutrophil oxidative
metabolism by leukotriene B4
JC Gay, JK Beckman, AR Brash, JA Oates and JN Lukens
Leukotriene B4 (LTB4) is a potent primary stimulator of neutrophil
chemotaxis, aggregation, and degranulation and induces superoxide
production at higher concentrations. In order to determine whether LTB4
modulates neutrophil responses to oxidative stimuli, human neutrophils
(PMNs) were incubated with LTB4 prior to stimulation with f-Met-Leu-Phe
(fMLP, 10(-7) mol/L), opsonized zymosan (OZ, 250 micrograms/mL), or phorbol
myristate acetate (PMA, 32 nmol/L). Superoxide (O2-) production by
stimulated PMNs was assessed by the superoxide dismutase-inhibitable
reduction of cytochrome c. LTB4 alone did not stimulate O2- production in
concentrations below 10(-7) mol/L and had no effect on the O2- assay. In
the concentration range of 10(-12) to 10(-8) mol/L, LTB4 did not alter O2-
release induced by OZ or PMA. In contrast, LTB4-treated cells demonstrated
enhanced O2- production following exposure to fMLP, and in the presence of
10 nmol/LLTB4, generated 180% +/- 41% of O-2 quantities produced by control
cells (n = 23). Enhancement was LTB4 dose-dependent, was maximal in the
range of 1 to 10 nmol/L LTB4, was not reversed by removal of the lipid from
the medium prior to fMLP stimulation, and was not dependent on the presence
of Ca++ or Mg++ in the suspending medium. Chemiluminescence of
fMLP-stimulated neutrophils was increased to 323% of controls in
neutrophils preincubated with 10 nmol/L LTB4. Unlike augmentation of
oxidative responses to fMLP seen with other degranulating stimuli,
enhancement by LTB4 was not correlated with an increase in 3H-fMLP receptor
binding. These results indicate that, in addition to its primary effects on
neutrophil function, LTB4 modulates PMN oxidative responses to the
chemotactic peptide and, thus, may amplify the release of oxygen
metabolites at inflammatory foci.
Volume 64,
Issue 4,
pp. 780-785,
10/01/1984
Copyright © 1984 by The American Society of Hematology
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