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Early gene expression changes with rush immunotherapy.doc

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Early gene expression changes with rush immunotherapy

Davisetal.ClinicalandMolecularAllergy2011,9:12 /content/9/1/12 CMA RESEARCH OpenAccess Earlygeneexpressionchangeswithrush immunotherapy LaurieSDavis1*,SumitBhutani2,SherryRidzBarnett2andDavidAKhan2 Abstract Background:ToexaminewhetherwholegenomeexpressionprofilingcouldrevealchangesinmRNAexpression ofperipheralbloodmononuclearcells(PBMC)fromallergicpatientsundergoingrushimmunotherapy(RIT)that mightbemanifestwithinthefirstfewmonthsoftreatment. Methods:Forthisstudy,PBMCfromthreeallergicpatientsundergoingRITwereassessedatfourtimepoints:prior toRIT,at1weekand7weekpost-RIT,duringbuild-upandat4months,afterestablishmentofamaintenance dose.PBMCmRNAgeneexpressionchangesovertimeweredeterminedbyoligonucleotidemicroarraysusingthe IlluminaHuman-6BeadChipPlatform,whichsimultaneouslyinterrogatesexpressionprofilesof47,000transcripts. Differentiallyexpressedgeneswereidentifiedusingwell-establishedstatisticalanalysisformicroarrays.Inaddition, weanalyzedperipheralbloodbasophilhigh-affinityIgEreceptor(FcepsilonRI)expressionandT-regulatorycell frequencyasdetectedbyexpressionofCD3+CD4+CD25brightcellsateachtimepointusingflowcytometry. Results:Incomparingtheinitial2timepointswiththefinal2timepointsandanalyzingforgeneswith≥1.5-fold expressionchange(plessthanorequalto0.05,BH-FDR),weidentified507transcripts.Ata2-foldchange(pless thanorequalto0.05,BH-FDR),wefound44transcripts.Ofthese,28wereup-regulatedand16weredown- regulatedgenes.Fromthesedatasets,wehaveidentifiedchangesinimmunologicallyrelevantgenesfromboth theinnateandadaptiveresponsewithupregulationofexpressedgenesformoleculesincludingIL-1b,IL-8,CD40L, BTKandBCL6.Atthe4monthtimepoint,wenotedadownwardtrendinFcepsilonRIexpressionineachofthe threepatientsandincreasedallergen-specificIgG4levels.NochangewasseeninthefrequencyofperipheralT- regulatorycellsexpressedoverthefourtimepoints. Conclusions:Weobservedsignificantchangesingeneexpressionearlyinperipheralbloodsamplesfromallergic patientsundergoingRIT.Moreover,serumlevelsforallergenspecificIg