An “Escape Clock” for Estimating the Turnover of SIV DNA in Resting CD4+ T Cells 英文参考文献.docVIP

An “Escape Clock” for Estimating the Turnover of SIV DNA in Resting CD4+ T Cells 英文参考文献.doc

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An“EscapeClock”forEstimatingtheTurnoverofSIVDNAinRestingCD4TCells英文参考文献

An‘‘EscapeClock’’forEstimatingtheTurnoverofSIV DNAinRestingCD4+TCells JeanetteReece1,JankaPetravic2,MehalaBalamurali2,LiyenLoh1,ShayaranaGooneratne1,RobDe Rose1,StephenJ.Kent1.*,MilesP.Davenport2. * 1Department ofMicrobiology andImmunology,UniversityofMelbourne,Parkville, Victoria, Australia, 2Centre forVascularResearch, UniversityofNew SouthWales, Sydney,NewSouthWales,Australia Abstract Persistence of HIV DNA presents a major barrier to the complete control of HIV infection under current therapies. Most studiessuggestthatcellswithlatentlyintegratedHIVdecayveryslowlyundertherapy.However,itismuchmoredifficultto studytheturnoverandpersistenceofHIVDNAduringactiveinfection.Wehavedevelopedan‘‘escapeclock’’approachfor measuringtheturnoverofHIVDNAinrestingCD4+Tcells.Thisapproachstudiesthereplacementofwild-type(WT)SIV DNApresentinearlyinfectionbyCTLescapemutant(EM)strainsduringlaterinfection.Usingastrain-specificrealtimePCR assay,wequantifiedtherelativeamountsofWTandEMstrainsinplasmaSIVRNAandcellularSIVDNA.Thuswecantrack theformationandturnoverofSIVDNAinsortedrestingCD4+Tcells.WestudiedserialplasmaandPBMCsamplesfrom20 SIV-infectedMane-A*10positivepigtailmacaquesthathaveasignatureGagCTLescapemutation.Inanimalswithlowviral load,WTviruslaiddownearlyininfectionisextremelystable,andthedecayofthisWTspeciesisveryslow,consistentwith findingsinsubjectsonanti-retroviralmedications.However,duringactive,highlevelinfection,mostSIVDNAinrestingcells was turning over rapidly, suggesting a large pool of short-lived DNA produced by recent infection events. Our results suggestthat,inordertoreducetheformationofastablepopulationofSIVDNA,itwillbeimportanteithertointervenevery earlyorinterveneduringactivereplication. Citation: Reece J, Petravic J, Balamurali M, Loh L, Gooneratne S, et al. (2012) An ‘‘Escape Clock’’ for Estimating the Turnover of SIV DNA in Resting CD4+ T Cells.PLoSPathog8(4):e1002615.doi:10.1371/journal.ppat.1002615 Editor:GuidoSilvestri,EmoryUniversity,UnitedStatesofAmeric

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