Non-Opsonic Phagocytosis of Legionella pneumophila by Macrophages Is Mediated by Phosphatidylinositol 3-Kinase 英文参考文献.docVIP

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Non-Opsonic Phagocytosis of Legionella pneumophila by Macrophages Is Mediated by Phosphatidylinositol 3-Kinase 英文参考文献.doc

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Non-Opsonic Phagocytosis of Legionella pneumophila by Macrophages Is Mediated by Phosphatidylinositol 3-Kinase 英文参考文献

Non-OpsonicPhagocytosisofLegionellapneumophila byMacrophagesIsMediatedbyPhosphatidylinositol 3-Kinase SouvenirD.Tachado¤a,MustaphaM.Samrakandi¤b,JeffreyD.Cirillo* DepartmentofMicrobialandMolecularPathogenesis,TexasAMHealthScienceCenter,CollegeStation,Texas,UnitedStatesofAmerica Abstract Background:Legionellapneumophila,isanintracellularpathogenthatcausesLegionnaires’diseaseinhumans,apotentially lethalpneumonia.L.pneumophilahastheabilitytoenterandreplicateinthehostandisessentialforpathogenesis. Methodology/PrincipalFindings:Phagocytosiswasmeasuredbycellinvasionassays.ConstructionofPI3KmutantbyPCR cloningandexpressionofdominantnegativemutantwasdetectedbyWesternblot.PI3Kactivitywasmeasuredby32 Plabeling anddetectionofphospholipidsproductsbythinlayerchromatography.InfectionofmacrophageswithvirulentL.pneumophila stimulated the formation of phosphatidylinositol 3-phosphate (PIP3), a phosphorylated lipid product of PI3K whereas two structurallydistinctphosphatidylinositol3kinase(PI3K)inhibitors,wortmanninandLY294002,reducedL.pneumophilaentry intomacrophagesinadose-dependentfashion.Furthermore,PI3KactivationledtoAktstimulation,aserine/threoninekinase, whichwasalsoinhibitedbywortmanninandLY294002.Incontrast,PI3KandproteinkinaseB(PKB/Akt)activitieswerelowerin macrophagesinfectedwithanavirulentbacterialstrain.OnlyvirulentL.pneumophilaincreasedlipidkinaseactivitypresentin immunoprecipitates of the p85a subunit of class I PI3K and tyrosine phosphorylated proteins. In addition, macrophages expressingaspecificdominantnegativemutantofPI3KreducedL.pneumophilaentryintothesecells. Conclusion/Significance: Entry of L. pneumophila is mediated by PI3K/Akt signaling pathway. These results suggest an important role for PI3K and Akt in the L. pneumophila infection process. They point to possible novel strategies for underminingL.pneumophilahostuptakeandreducingpathogenesisofLegionnaires’disease. Citation: Tachado SD, Samrakandi MM, Cirillo JD (2008) Non-Opsonic Phagocytosis of Legionell