英语国际会议

.,1,MacrophagesfrompatientswithatopicdermatitisshowareducedCXCL10expressioninresponsetostaphylococcala-toxin.,.,2,CONTENTS,Introduction,Results,MaterialsandMethods,Discussion,.,3,background,.,4,Keywords,atopicdermatitis;(特应性皮炎)chemokines;（趋化因子）humanmacrophages;（人巨噬细胞）IP-10/CXCL10;MDC/CCL22;psoriasis;（牛皮癣；银屑病）Staphylococcusaureus;（金黄色葡萄球菌）-toxin.（-毒素）,.,5,Background:,Patientswithatopicdermatitis(AD)andpsoriasisarefrequentlycolonizedwithStaphylococcusaureus(S.aureus).one-thirdofthemproducing-toxin,whichiscor-relatedwiththeseverityofeczemainAD.Distinctexpressionofchemokine(C-Cmotif)ligand(配体CCL)andchemokine(C-X-Cmotif)ligand(配体CXCL)chemokineshasbeendocumentedinbothdiseases.theeffectsofa-toxinonhumanmacrophagesinpatientswithADandpsoriasis?,.,6,Presentstudy,Recentstudiesdemonstratedthatinfiltrationofinflammatorycellsintotissuesisregulatedbychemokines.Twomainsubfamilies,recentlyrenamedchemokine(C-Cmotif)ligand(CCL)andchemokine(C-X-Cmotif)ligand(CXCL)chemo-kines,havebeendistinguished.SeveralstudieshavesuggestedacrucialinvolvementofCCLchemokinesininflammation.ChemokinesandtheirreceptorsareimplicatedinthedevelopmentofsymptomsofADandpsoriasis.CXCL10playsaroleinpathogenesisofpsoriasis.,.,7,Purpose,Theeffectsof-toxinonhumanmacrophagesstillremainunclear.Weshowhereforthefirsttimethat-toxininducestheTh1-relatedchemokineCXCL10inhumanmacrophages.westudiedtheeffectsof-toxinonTh1-andTh2-relatedchemokinesinmacrophagesfrompatientswithADandpsoriasiswheretheintrinsicabnormalanddifferentchemokinesproductionprofileiswelldefined.,.,8,MaterialsandMethods,MaterialsandMethods,Patients,Cellisolationandculture,Statisticalanalysis,CytokineassessmentbyELISA,Westernblot,.,9,Methods,macrophage-derivedchemokine(MDC)/CCL22,Cellsurfacemarkersexpressionandchemotaxis,IFN-c-inducedproteinof10-kDa(IP-10)/CXCL10,qRTPCRorELISA,flowcytometryBoydenchambertechnique,.,10,Patients,healthydonorsandpatientswithADorpsoriasis,Cellisolationandculture,Peripheralbloodmononuclearcells(PBMCs)wereisolatedbyLymphoprepdensity-gradientcentrifugationfromhealthydonorsandfrompatientswithADandpsoriasis,Chemotaxisassay,Thechemotacticactivityofsupernatantsfroma-toxinstimulatedmacrophagesonlymphocytes(CD4+Tcells)wasdeterminedusingaBoydenchambertechnique,.,11,Westernblot,macrophageswereprestimulatedwitha-toxin(100ng/ml),IFN-c(10ng/ml),TNF-a(10ng/ml)orLPS.CellextractsweresubjectedtoWesternblotanalysis,CytokineassessmentbyELISA,Cell-freeculturesupernatantswereharvestedandanalysedforCXCL10orCCL22,usingacommerciallyavailableenzyme-linkedimmunosorbentassay(ELISA)system.,.,12,Results,OneInductionofIP-10/CXCL10bystaphylococcala-toxininhumanmacrophages,ThreeLoweffectofa-toxinonCXCL10induction(Th1-relatedchemokine)inmacrophagesfrompatientswithAD,Twoa-Toxin-stimulatedmacrophagescouldinducethemigrationofhumanCD4+TcellsviaCXCR3,FourEffectofa-toxinonMDCproduction(Th2-relatedchemokine)inmacrophagesfrompatientswithchronicinammatoryskindiseases,.,13,InductionofIP-10/CXCL10bystaphylococcala-toxininhumanmacrophages,MicroarrayanalysisrevealedthatCXCL10wasthemoststronglyup-regulatedgeneinmacrophages.Anincreasedexpressionwasnotobservedforotherchemokines.Themeanratioforallthesechemokineswasbetween0.2and2whencomparingnotstimulatedwitha-toxin-stimulatedmacrophages.,.,14,Figure1Inductionofchemokine(C-X-Cmotif)ligand(CXCL)10followinga-toxinstimulationinhumanmacrophagesatthemRNAlevel.,Figure2Inductionofchemokine(C-X-Cmotif)ligand(CXCL)10followinga-toxinstimulationinhumanmacrophagesattheproteinlevel.,.,15,Figure3Punchbiopsies(3mm)fromhealthyindividualswerelefteitherunstimulated(A)orstimulatedwitha-toxin(100ng/ml)(B)orIFN-c(100ng/ml)(C)for24hat37C.5-lmparafnsectionswerestainedforCXCL10alongwithappropriateisotypeaswellasCD68.,.,16,a-Toxin-stimulatedmacrophagescouldinducethemigrationofhumanCD4+TcellsviaCXCR3,Bothinhibitors(anti-IP-10antibody&anti-CXCR3antibody)abrogatedthemigrationinducedbya-toxin-treatedmacrophagesupernatants.a-ToxinincellculturemediumwithoutmacrophagesdidnotdirectlyinducethemigrationofhumanCD4+lymphocytes.,.,17,Figure4Chemotacticactivityofa-toxin-stimulatedmacrophagesandinhibitionofchemokine(C-X-Cmotif)ligand(CXCL)10-inducedchemotaxisusinganeutralizinganti-CXCL10oranti-CXCR3Ab.,.,18,Loweffectofa-toxinonCXCL10induction(Th1-relatedchemokine)inmacrophagesfrompatientswithAD,a-toxininducedsignicantlylowerlevelsofCXCL10expressionorsecretioninpatientswithADcomparedwithhealthycontrolsaswellaspsoriasisatalltimepointsanddosestested.,.,19,Figure5Macrophagesfrompatientswithatopicdermatitis(AD)andpsoriasis(PSO)aswellasfromhealthycontrols(HC)werelefteitherunstimulated(mediumcontrol)orstimulatedwitha-toxinasindicatedorwithIFN-c(10ng/ml)orTNF-a(20ng/ml),respectively,fortheindicatedperiodsoftime.,.,20,Effectofa-toxinonMDCproduction(Th2-relatedchemokine)inmacr