Regulation of human eosinophil NADPH oxidase activity: A central role for PKCδ

Abstract Eosinophils play a primary role in the pathophysiology of asthma. In the lung, the activation state of the infiltrating eosinophils determines the extent of tissue damage. Interleukin‐5 (IL‐5) and leukotriene B 4 (LTB 4 ) are important signaling molecules involved in eosinophil recruitment and activation. However, the physiological processes that regulate these activation events are largely unknown. In this study we have examined the mechanisms of human eosinophil NADPH oxidase regulation by IL‐5, LTB 4 , and phorbol ester (PMA). These stimuli activate a Zn 2+ ‐sensitive plasma membrane proton channel, and treatment of eosinophils with Zn 2+ blocks superoxide production. We have demonstrated that eosinophil intracellular pH is not altered by IL‐5 activation of NADPH oxidase. Additionally, PKCδ inhibitors block PMA, IL‐5 and LTB 4 mediated superoxide formation. Interestingly, the PKCδ‐selective inhibitor, rottlerin, does not block proton channel activation by PMA indicating that the oxidase and the proton conductance are regulated at distinct phosphorylation sites. IL‐5 and LTB 4 , but not PMA, stimulated superoxide production is also blocked by inhibitors of PI 3‐kinase indicating that activation of this enzyme is an upstream event common to both receptor signaling pathways. Our results indicate that the G‐protein‐coupled LTB 4 receptor and the IL‐5 cytokine receptor converge on a common signaling pathway involving PI 3‐kinase and PKCδ to regulate NADPH oxidase activity in human eosinophils. © 2001 Wiley‐Liss, Inc.