Fluorescent Cell Barcoding as a Tool to Assess the Age-Related Development of Intracellular Cytokine Production in Small Amounts of Blood from Infants 英文参考文献.docVIP

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Fluorescent Cell Barcoding as a Tool to Assess the Age-Related Development of Intracellular Cytokine Production in Small Amounts of Blood from Infants 英文参考文献.doc

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Fluorescent Cell Barcoding as a Tool to Assess the Age-Related Development of Intracellular Cytokine Production in Small Amounts of Blood from Infants 英文参考文献

FluorescentCellBarcodingasaTooltoAssessthe Age-RelatedDevelopmentofIntracellularCytokine ProductioninSmallAmountsofBloodfromInfants JoseStam1*.,WayelAbdulahad2.,MinkeG.Huitema2,CarolineRoozendaal3,PieterC.Limburg3, MargrietvanStuijvenberg1,ElisabethH.Scho¨lvinck1 1Beatrix Children’s Hospital, University Medical Centre Groningen, Groningen, the Netherlands, 2Department of Rheumatology and Clinical Immunology, University MedicalCentreGroningen,Groningen,theNetherlands,3DepartmentofLaboratoryMedicine,UniversityMedicalCentreGroningen,Groningen,theNetherlands Abstract FluorescentCellBarcoding(FCB)isaflowcytometrictechniquewhichhasbeenusedforassessingsignalingproteins.This FCBtechniquehasthepotentialtobeappliedinothermultiparameteranalyses.Sincedataonantigen(Ag)-specificT-cell immuneresponses,likeintracellularcytokineproduction,arestilllackingininfantsbecauselimitedbloodvolumescanbe obtainedforanalysis,theFCBtechniquecouldbeveryusefulforthispurpose.Theobjectivesofthisstudyweretomodify theFCBmethodtobeabletomeasuremultipleAg-specificcytokinereponsesinT-cellsuponsimultaneousstimulationby variousantigensandmitogensinsmallamountsofbloodandtoinvestigatethecytokinepatternofT-cellsubsetsinhealthy infantsagedsixandtwelvemonths.Bloodsamples,collectedfrom20healthyinfantsagedsixandtwelvemonths,were stimulated in vitro with the antigens: phorbol-myristate-acetate (PMA), purified-protein-derivative (PPD), Tetanus-toxoid (TT), Staphylococcal-enterotoxin-B (SEB), and phytohemagglutinin (PHA). Each stimulus was barcoded by labelling with differentintensitiesoffluorescentcellbarcoding(FCB)markers.Intracellularproductionofinterleukin-2,interferon-gamma, andtumornecrosisfactor-alphawasmeasuredsimultaneouslyinjustonebloodsampleof600mlwholeblood.Significant + age-relateddifferencesincytokineproductionwereshownforPMA,PHA,andTTinCD4 T-cells,andforPMA,PHA,SEB, and TT in CD8+ T-cells. The intracellular cytokine production by CD4+ and CD8+ T-cells was higher at twelve months comparedtosix