Bacterial Thymidine Kinase as a Non-Invasive Imaging Reporter for Mycobacterium tuberculosis in Live Animals 英文参考文献.docVIP

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Bacterial Thymidine Kinase as a Non-Invasive Imaging Reporter for Mycobacterium tuberculosis in Live Animals 英文参考文献.doc

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Bacterial Thymidine Kinase as a Non-Invasive Imaging Reporter for Mycobacterium tuberculosis in Live Animals 英文参考文献

BacterialThymidineKinaseasaNon-InvasiveImaging ReporterforMycobacteriumtuberculosisinLiveAnimals StephanieL.Davis1,2,NicholasA.Be1,GyanuLamichhane1,3,SridharNimmagadda4,MartinG.Pomper4, WilliamR.Bishai1,3,SanjayK.Jain1,2 * 1Center for Tuberculosis Research, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America, 2Department of Pediatrics, Johns HopkinsUniversitySchoolofMedicine,Baltimore,Maryland,UnitedStatesofAmerica,3DepartmentofMedicine,JohnsHopkinsUniversitySchoolofMedicine,Baltimore, Maryland,UnitedStatesofAmerica,4DepartmentofRadiology,JohnsHopkinsUniversitySchoolofMedicine,Baltimore,Maryland,UnitedStatesofAmerica Abstract Background: Bacteria can be selectively imaged in experimentally-infected animals using exogenously administered 1- (29deoxy-29-fluoro-b-D-arabinofuranosyl)-5-[125I]-iodouracil ([125I]-FIAU), a nucleoside analog substrate for bacterial thymidine kinase (TK). Our goal was to use this reporter and develop non-invasive methods to detect and localize Mycobacteriumtuberculosis. Methodology/PrincipalFindings: WeengineeredaM.tuberculosisstrainwithchromosomallyintegratedbacterialTKunder thecontrolofhsp60-astrongconstitutivemycobacterialpromoter.[125I]FIAUuptake,antimicrobialsusceptibilitiesandin vivogrowthcharacteristicswereevaluatedforthisstrain.Usingsinglephotonemissioncomputedtomography(SPECT),M. tuberculosis Phsp60 TK strain was evaluated in experimentally-infected BALB/c and C3HeB/FeJ mice using the thigh inoculation or low-dose aerosol infection models. M. tuberculosis Phsp60 TK strain actively accumulated [125I]FIAU in vitro. GrowthcharacteristicsoftheTKstrainandsusceptibilitytocommonanti-tuberculousdrugsweresimilartothewild-type parentstrain.M.tuberculosisPhsp60 TKstrainwasstableinvivoandSPECTimagingcoulddetectandlocalizethisstrainin bothanimalmodelstested. Conclusion:Wehavedevelopedanoveltoolfornon-invasiveassessmentofM.tuberculosisinliveexperimentally-infected animals. This tool will allow real-tim