Leukotriene B4 activates the NADPH oxidase in eosinophils by a pertussis toxin-sensitive mechanism that is largely independent of arachidonic acid mobilization

Mark A. Lindsay, Rosie S. Perkins, Peter J. Barnes, Mark A. Giembycz

Research output: Contribution to journalArticlepeer-review

Abstract

Experiments were designed to investigate whether leukotriene (LTB4) receptors can couple directly to phospholipase A2 (PLA2) in guinea pig eosinophils and the role of endogenous arachidonic acid (AA) in LTB4- induced activation of the NADPH oxidase. LTB4 (EC50 ~ 16 nM) and AA (EC50 ~ 6 μM) generated hydrogen peroxide (H2O2) in a concentration- dependent manner and at an equivalent maximum rate (5-6 nmol/min/106 cells). LTB4 stimulated PLA2 over a similar concentration range that activated the NADPH oxidase, although kinetic studies revealed that the release of [3H]AA (t 1/4 ~ 2 s) preceded H2O2 generation (t 1/4 > 30 s). Pretreatment of eosinophils with pertussis toxin abolished the increase in inositol(1,4,5)trisphosphate mass, [Ca2+](c), [3H]AA release, and H2O2 generation evoked by LTB4. Qualitatively identical results were obtained in eosinophils in which phospholipase C (PLC) was desensitized by 4β-phorbol 12,13-dibutyrate with the exception that [3H]AA release was largely unaffected. Additional studies performed with the protein kinase C inhibitor, Ro 31-8220, and under conditions in which Ca2+ mobilization was abolished, provided further evidence that LTB4 released [3H]AA independently of signal molecules derived from the hydrolysis ofphosphatidylinositol(4,5)bisphosphate by PLC. Pretreatment of eosinophils with the PLA2 inhibitor, mepacrine, abolished LTB4-induced [3H]AA release at a concentration that inhibited H2O2 by only 36%. Collectively, the results of this study indicate that agonism of LTB4 receptors on guinea pig eosinophils mobilizes AA by a mechanism that does not involve the activation of PLC. In addition, although LTB4 effectively stimulated PLA2, a central role for AA in the activation of the NADPH oxidase was excluded.

Original languageEnglish
Pages (from-to)4526-4534
Number of pages9
JournalJournal of Immunology
Volume160
Issue number9
Publication statusPublished - 1 May 1998

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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