Protein Kinase A Regulates beta-2 Integrin Avidity Activation and Subsequent Neutrophil Activation via Modulation of Myosin Light Chain Kinase.

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dc.contributor.advisor Dr. Robert B. Rose, Committee Member en_US
dc.contributor.advisor Dr. Samuel L. Jones, Committee Co-Chair en_US
dc.contributor.advisor Dr. Scott M. Laster, Committee Member en_US
dc.contributor.advisor Dr. Edward A. Havell, Committee Member en_US
dc.contributor.advisor Dr. Wayne A. Tompkins, Committee Co-Chair en_US
dc.contributor.author Chilcoat, Clayton Douglas en_US
dc.date.accessioned 2010-04-02T19:09:02Z
dc.date.available 2010-04-02T19:09:02Z
dc.date.issued 2006-04-11 en_US
dc.identifier.other etd-03312005-093042 en_US
dc.identifier.uri http://www.lib.ncsu.edu/resolver/1840.16/5175
dc.description.abstract β2 integrins are adhesion molecules on the surface of neutrophils. Avidity activation of β2 integrins includes transportation of pre-formed integrins to the cell surface and a conformational change in the integrin to a high-binding state. Upon binding ligand, β2 integrins initiate a signaling cascade that results in activation of the neutrophil to a pro-inflammatory state, and the inhibition of this signal can prevent further activation of the neutrophil. cAMP and it effector protein kinase A (PKA) exert a generally inhibitory effect upon neutrophil activation. PKA has been shown to inactivate myosin light chain kinase (MLCK). Myosin light chain (MLC) phosphorylation is crucial for actin-myosin complex formation, which is required for stability and contraction of the actin cytoskeleton in neutrophils as well as β2 integrin-dependent adhesion. We hypothesize that the inhibitory effect of PKA upon neutrophils is due to inhibition of β2 integrin avidity activation resulting in the subsequent inhibition of neutrophil activation. Furthermore we hypothesize that the effect of PKA upon β2 integrin avidity activation is mediated through PKA's effect upon MLCK. We demonstrate that inhibition of PKA induces β2 integrin-dependent adhesion and that augmentation of cAMP prevented β2 integrin-dependent adhesion and subsequent respiratory burst activity. Further, we demonstrate via flow cytometric detection of antibodies directed against β2 integrins that pharmacologic inhibition of PKA activity results in overall increased β2 integrin expression on the neutrophil surface, as well as increased expression of the activated form of the integrin. This upregulation and activation of β2 integrins due to inhibition of PKA is abolished by pharmacologic MLCK inhibition. Inhibition of MLCK also blocked β2 integrin-dependent neutrophil adhesion achieved by inhibition of PKA, as well as neutrophil migration along towards a PKA inhibitor. These findings demonstrate that PKA regulation of β2 integrin affinity activation and subsequent neutrophil activation is via an MLCK-dependent pathway. en_US
dc.rights I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to NC State University or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. en_US
dc.subject MLCK en_US
dc.subject integrin en_US
dc.subject myosin en_US
dc.subject PKA en_US
dc.subject PMN en_US
dc.subject MLC en_US
dc.subject neutrophil en_US
dc.title Protein Kinase A Regulates beta-2 Integrin Avidity Activation and Subsequent Neutrophil Activation via Modulation of Myosin Light Chain Kinase. en_US
dc.degree.name PhD en_US
dc.degree.level dissertation en_US
dc.degree.discipline Immunology en_US


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