NSOMQD-Based Direct Visualization of CD3-Induced and CD28-Enhanced Nanospatial Coclustering of TCR and Coreceptor in Nanodomains in T Cell Activation 英文参考文献.docVIP

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NSOMQD-Based Direct Visualization of CD3-Induced and CD28-Enhanced Nanospatial Coclustering of TCR and Coreceptor in Nanodomains in T Cell Activation 英文参考文献.doc

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NSOMQD-Based Direct Visualization of CD3-Induced and CD28-Enhanced Nanospatial Coclustering of TCR and Coreceptor in Nanodomains in T Cell Activation 英文参考文献

NSOM/QD-BasedDirectVisualizationofCD3-Induced andCD28-EnhancedNanospatialCoclusteringofTCR andCoreceptorinNanodomainsinTCellActivation LiyunZhong1.,GuchengZeng1.,XiaoxuLu2,RichardC.Wang1,GuangmingGong1,LinYan1 ,Dan Huang1,ZhengW.Chen1* 1Department of Microbiology and Immunology, Center for Primate Biomedical Research, University of Illinois College of Medicine, Chicago, Illinois, United States of America,2SchoolforInformationandOptoelectronicEngineering,SouthChinaNormalUniversity,Guangzhou,Guangdong,China Abstract Directmolecularimagingofnano-spatialrelationshipbetweenTcellreceptor(TCR)/CD3andCD4orCD8co-receptorbefore andafteractivationofaprimaryTcellhasnotbeenreported.Wehaverecentlyinnovatedapplicationofnear-fieldscanning optical microscopy (NSOM) and immune-labeling quantum dots (QD) to image Ag-specific TCR response during in vivo clonal expansion, and now up-graded the NSOM/QD-based nanotechnology through dipole-polarization and dual-color imaging. Using this imaging system scanning cell-membrane molecules at a best-optical lateral resolution, we demonstratedthatCD3,CD4orCD8moleculesweredistinctlydistributedassingleQD-boundmoleculesornano-clusters equivalentto2–4QDfluorescence-intensity/sizeoncell-membraneofun-stimulatedprimaryTcells,and,6–10%ofCD3 wereco-clusteringwithCD4orCD8as70–110nmnano-clusterswithoutformingnano-domains.TheligationofTCR/CD3 onCD4orCD8TcellsledtoCD3nanoscaleco-clusteringorinteractionwithCD4orCD8co-receptorsforming200–500nm nano-domainsor.500nmmicro-domains.Suchnano-spatialco-clusteringofCD3andCD4orCD3andCD8appearedto beanintrinsiceventofTCR/CD3ligation,notpurelylimitedtoMHCengagement,andbedrivenbyLckphosphorylation. Importantly, CD28 co-stimulation remarkably enhanced TCR/CD3 nanoscale co-clustering or interaction with CD4 co- receptorwithinnano-ormicro-domainsonthemembrane.Incontrast,CD28co-stimulationdidnotenhanceCD8clustering orCD3–CD8co-clusteringinnano-domainsalthoughitincreasedmolecularnumberanddensityofCD3clusteringinthe enlarged