Regulatorystatus:Forresearchuseonly
Capacity:for100tests(1)Overview
TheIFN-γSecretionAssaywasdevelopedforthesensitivedetectionofhumanIFN-γ-secretingcells.
Details
Backgroundinformation
IFN-γ(interferon-gamma)ispredominantlysecretedbyactivatedCD8+andCD4+memoryandeffectorTcellsandbyNKcells.Itismainlyinvolvedintheregulationofinflammatoryimmuneresponses.TheseTH1typesofimmunemechanismsareeffectiveagainstintracellularpathogensandtumors.IFN-γ-secretingTcellscanalsobeinvolvedinimmunologicaldisorders,suchasautoimmunereactions.
Downstreamapplications
Virus-specificTcellswereinvestigatedafterstimulationwithpeptidesorproteinsderivedfrominfluenzavirus1,CMV2,3,EBV4,5,6,HIV5,7,8,9,10,HBV11,andADV24.
Virus-specificTcellswereexpandedinvitro1,2,3,5,6,9showinghighlyspecificandveryefficientkillingoftargetcellsandhavebeenanalyzedforTCRclonotypes8,10.
TheIFN-γSecretionAssaywasusedfortheisolationandanalysisofantigen-specificTcellsfromPBMCsafterstimulationwithTetanusToxoid1,minorhistocompatibilityantigens,andtumorantigens12,13,14,15.Theassaywasalsousedtopurifyandanalyzetumor-specificTcellsfromTcelllines13,14,fortheisolationoffunctionalantigen-specific,IFN-γ-secretingTcellsreactingtoothertumorantigens,e.g.SSX14,CEA16,orHER217fromPBMCsorTILs(tumorinfiltratinglymphocytes)14,18.
TheIFN-γSecretionAssaywasusedtocounterstainpeptide-MHC-tetramer-labeledMelanA-specificCD8+Tcellstoanalyzefunctionalityofthetetramer-positivecells.12,15
TheIFN-γSecretionAssaywasalsousedforisolationandfunctionalcharacterizationofallergen-specificTcells.
21Fur
Thermore,theIFN-γSecretionAssaywasusedforepitopemappingofMHCclassIIpeptides.
19IFN-γ-secretinghumanNKcellswereisolatedusingtheIFN-γSecretionAssay.20IFN-γSecretionAssayreagentswerereportedtocross-reactwithchimpanzeecells22butnotwithrhesusmacaquecells.
TheIFN-γSecretionAssaycanalsobeusedfortwo-colorcytokineanalysis9andallowscounterstainingofpeptide-MHC-tetramer-labeledcells12,15.Itcanalsobecombinedwithflowcytometricproliferationassays23.