title: Critical role of proline-rich tyrosine kinase 2 in reversion of the adhesion-mediated suppression of reactive oxygen species generation by human neutrophils.
abstract: Neutrophils act as the first line of innate immune defense against invading microorganisms during infection and inflammation. The tightly regulated production of reactive oxygen species (ROS) through activation of NADPH oxidase is a major weapon used by neutrophils and other phagocytic leukocytes to combat such pathogens. Cellular adhesion signals play important physiological roles in regulating the activation of NADPH oxidase and subsequent ROS formation. We previously showed that the initial suppression of the oxidase response of chemoattractant-stimulated adherent neutrophils is mediated via inhibition of Vav1-induced activation of the NADPH oxidase regulatory GTPase Rac2 by adhesion signals. In this study we show that prior exposure of neutrophils to a number of cytokines and inflammatory mediators, including TNF-alpha, GM-CSF, and platelet-activating factor, overcomes the adhesion-mediated suppression of ROS formation. Proline-rich tyrosine kinase 2 (pyk2) activity is enhanced under these conditions, correlating with the restoration of Vav1 and Rac2 activities. Both dominant negative pyk2 and a pyk2-selective inhibitor prevented restoration of ROS production induced by TNF-alpha, GM-CSF, and platelet-activating factor, and this loss of pyk2 activity resulted in decreased Vav1 tyrosine phosphorylation and subsequent Rac2 activation. Our studies identify pyk2 as a critical regulatory component and a molecular switch to overcome the suppression of leukocyte oxidant generation by cell adhesion. This activity constitutes a mechanism by which cytokines might lead to rapid elimination of invading pathogens by adherent neutrophils under normal conditions or enhance tissue damage in pathological states.
authors.affiliation: Department of Immunology, The Scripps Research Institute, La Jolla, CA 92037, USA. tzhao@scripps.edu
authors.forename: Tieming
authors.initials: T
authors.lastname: Zhao
authors.forename: Gary M
authors.initials: GM
authors.lastname: Bokoch
mesh_terms: D002448:Cell Adhesion / Q000276:immunology; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D002630:Chemotactic Factors / Q000494:pharmacology; D015536:Down-Regulation / Q000276:immunology*; D004789:Enzyme Activation / Q000276:immunology; D051418:Focal Adhesion Kinase 2; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006801:Humans; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors / Q000378:metabolism; D009504:Neutrophils / Q000201:enzymology / Q000276:immunology* / Q000378:metabolism*; D016877:Oxidants / Q000037:antagonists & inhibitors / Q000502:physiology; D010766:Phosphorylation; D010972:Platelet Activating Factor / Q000502:physiology; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000378:metabolism / Q000502:physiology*; D011518:Proto-Oncogene Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D051567:Proto-Oncogene Proteins c-vav; D017382:Reactive Oxygen Species / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000502:physiology; D014443:Tyrosine / Q000378:metabolism; D015854:Up-Regulation / Q000276:immunology*; D020744:rac GTP-Binding Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D000097766:RAC2 GTP-Binding Protein
chemical_list: D018797:Cell Cycle Proteins; D002630:Chemotactic Factors; D051418:Focal Adhesion Kinase 2; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D009240:N-Formylmethionine Leucyl-Phenylalanine; D019255:NADPH Oxidases; D016877:Oxidants; D010972:Platelet Activating Factor; D017027:Protein Tyrosine Phosphatases; D011505:Protein-Tyrosine Kinases; D011518:Proto-Oncogene Proteins; D051567:Proto-Oncogene Proteins c-vav; D017382:Reactive Oxygen Species; D014409:Tumor Necrosis Factor-alpha; D014443:Tyrosine; D020744:rac GTP-Binding Proteins; D000097766:RAC2 GTP-Binding Protein; C496618:VAV1 protein, human