Negative Selection Assay Based on Stimulation of T Cell Receptor Transgenic Thymocytes with Peptide-MHC Tetramers 英文参考文献.docVIP
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Negative Selection Assay Based on Stimulation of T Cell Receptor Transgenic Thymocytes with Peptide-MHC Tetramers 英文参考文献
NegativeSelectionAssayBasedonStimulationofTCell
ReceptorTransgenicThymocyteswithPeptide-MHC
Tetramers
VasilyRybakin*,NicholasR.J.Gascoigne*
DepartmentofImmunologyandMicrobialSciences,TheScrippsResearchInstitute,LaJolla,California,UnitedStatesofAmerica
Abstract
Thymocytenegativeselectionisarequirementforthedevelopmentofselftolerance.Althoughitispossibletoassaythe
inductionofcelldeathinthymocytesinvitrousingantibodycross-linking,thisstimulusismuchstrongerthanthenormal
rangeofTcellreceptorligandsthatcouldbeencounteredduringnormaldevelopment.Signalinginthymocytesisfinely
balancedbetweenpositiveandnegativeselectionstimuli,whereanegativeselectingligandcanbeonlymarginallyhigher
affinitythanapositiveselectingligand.Wehavethereforedevelopedanassayfortheinductionofnegativeselectionthat
can distinguish such cases, and that is amenable to high-throughput analysis. The assay is based on the induction of
activated caspase 3 in thymocytes expressing adefined Tcell receptor, after stimulationwithMHC-peptide tetramers in
vitrofor24hoursorless.
Citation: Rybakin V, Gascoigne NRJ (2012) Negative Selection Assay Based on Stimulation of T Cell Receptor Transgenic Thymocytes with Peptide-MHC
Tetramers.PLoSONE7(8):e43191.doi:10.1371/journal.pone.0043191
Editor:JoseAlberola-Ila,OklahomaMedicalResearchFoundation,UnitedStatesofAmerica
ReceivedJune7,2012;AcceptedJuly20,2012;PublishedAugust10,2012
Copyright:?2012Rybakin,Gascoigne.Thisisanopen-accessarticledistributedunderthetermsoftheCreativeCommonsAttributionLicense,whichpermits
unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited.
Funding:ThisworkwassupportedbyNationalInstitutesofHealthgrantsAI073870,AI074074andGM065230.Thefundershadnoroleinstudydesign,data
collectionandanalysis,decisiontopublish,orpreparationofthemanuscript.
CompetingInterests:Theauthorshavedeclaredthatnocompetinginterestsexist.
*E-mail:vrybakin@(VR);ngascoigne@(NRJG)
Introduction
concentrations of anti-CD3 mimic positive s
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