Simple diagnosis of STAT1 gain‐of‐function alleles in patients with chronic mucocutaneous candidiasis
[摘要] CMCDisararecongenitaldisordercharacterizedbypersistentorrecurrentskin,nail,andmucosalmembraneinfectionscausedbyCandidaalbicans.HeterozygousGOFSTAT1mutationshavebeenshowntoconferADCMCDasaresultofimpaireddephosphorylationofSTAT1.WeaimedtoidentifyandcharacterizeSTAT1mutationsinCMCDpatientsandtodevelopasimplediagnosticassayofCMCD.GeneticanalysisofSTAT1wasperformedinpatientsandtheirrelatives.Themutationsidentifiedwerecharacterizedbyimmunoblotandreporterassayusingtransientgeneexpressionexperiments.Patientsˈleukocytesareinvestigatedbyflowcytometryandimmunoblot.SixGOFmutationswereidentified,threeofwhicharereportedforthefirsttime,thataffecttheCCDandDBDofSTAT1intwosporadicandfourmultiplexcasesin10CMCDpatientsfromJapan.Twoofthe10patientspresentedwithclinicalsymptomsatypicaltoCMCD,includingotherfungalandviralinfections,andthreepatientsdevelopedbronchiectasis.ImmunoblotanalysesofpatientsˈleukocytesshowedabnormallyhighlevelsofpSTAT1followingIFN‐γstimulation.Basedonthisfinding,weperformedaflowcytometry‐basedfunctionalanalysisofSTAT1GOFallelesusingIFN‐γstimulationandthetyrosinekinaseinhibitor,staurosporine.ThehigherlevelsofpSTAT1observedinprimaryCD14+cellsfrompatientscomparedwithcontrolcellspersistedandwereamplifiedbythepresenceofstaurosporine.Wedevelopedaflowcytometry‐basedSTAT1functionalscreeningmethodthatwouldgreatlyfacilitatethediagnosisofCMCDpatientswithGOFSTAT1mutations...
[发布日期] [发布机构]
[效力级别] [学科分类] 生理学
[关键词] CMCD;STAT1;GOF;pSTAT1 [时效性]
