英语国际会议PPT

1MacrophagesfrompatientswithatopicdermatitisshowareducedCXCL10expressioninresponsetostaphylococcala-toxin.IntroductionResultsMaterialsandMethods2Discussion3IntroductionkeywordspresentstudypurposebackgroundKeywords•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）•α-toxin.（α-毒素）4Background:•Patientswithatopicdermatitis(AD)andpsoriasisarefrequentlycolonizedwithStaphylococcusaureus(S.aureus).one-thirdofthemproducingα-toxin,whichiscor-relatedwiththeseverityofeczemainAD.•Distinctexpressionofchemokine(C-Cmotif)ligand(配体CCL)andchemokine(C-X-Cmotif)ligand(配体CXCL)chemokineshasbeendocumentedinbothdiseases.•theeffectsofa-toxinonhumanmacrophagesinpatientswithADandpsoriasis?5Presentstudy6Recentstudiesdemonstratedthatinfiltrationofinflammatorycellsintotissuesisregulatedbychemokines.Twomainsubfamilies,recentlyrenamedchemokine(C-Cmotif)ligand(CCL)andchemokine(C-X-Cmotif)ligand(CXCL)chemo-kines,havebeendistinguished.SeveralstudieshavesuggestedacrucialinvolvementofCCLchemokinesininflammation.ChemokinesandtheirreceptorsareimplicatedinthedevelopmentofsymptomsofADandpsoriasis.CXCL10playsaroleinpathogenesisofpsoriasis.Purpose7Theeffectsofα-toxinonhumanmacrophagesstillremainunclear.Weshowhereforthefirsttimethatα-toxininducestheTh1-relatedchemokineCXCL10inhumanmacrophages.westudiedtheeffectsofα-toxinonTh1-andTh2-relatedchemokinesinmacrophagesfrompatientswithADandpsoriasiswheretheintrinsicabnormalanddifferentchemokinesproductionprofileiswelldefined.MaterialsandMethods8MaterialsandMethodsPatientsCellisolationandcultureStatisticalanalysisCytokineassessmentbyELISAWesternblotMethods9macrophage-derivedchemokine(MDC)/CCL22Cellsurfacemarkers’expressionandchemotaxisIFN-c-inducedproteinof10-kDa(IP-10)/CXCL10qRT–PCRorELISAflowcytometryBoydenchambertechnique10PatientshealthydonorsandpatientswithADorpsoriasisCellisolationandculturePeripheralbloodmononuclearcells(PBMCs)wereisolatedbyLymphoprepdensity-gradientcentrifugationfromhealthydonorsandfrompatientswithADandpsoriasisChemotaxisassayThechemotacticactivityofsupernatantsfroma-toxinstimulatedmacrophagesonlymphocytes(CD4+Tcells)wasdeterminedusingaBoydenchambertechnique11Westernblotmacrophageswereprestimulatedwitha-toxin(100ng/ml),IFN-c(10ng/ml),TNF-a(10ng/ml)orLPS.CellextractsweresubjectedtoWesternblotanalysisCytokineassessmentbyELISACell-freeculturesupernatantswereharvestedandanalysedforCXCL10orCCL22,usingacommerciallyavailableenzyme-linkedimmunosorbentassay(ELISA)system.Results12OneInductionofIP-10/CXCL10bystaphylococcala-toxininhumanmacrophagesThreeLoweffectofa-toxinonCXCL10induction(Th1-relatedchemokine)inmacrophagesfrompatientswithADTwoa-Toxin-stimulatedmacrophagescouldinducethemigrationofhumanCD4+TcellsviaCXCR3FourEffectofa-toxinonMDCproduction(Th2-relatedchemokine)inmacrophagesfrompatientswithchronicinflammatoryskindiseasesInductionofIP-10/CXCL10bystaphylococcala-toxininhumanmacrophages13MicroarrayanalysisrevealedthatCXCL10wasthemoststronglyup-regulatedgeneinmacrophages.Anincreasedexpressionwasnotobservedforotherchemokines.Themeanratioforallthesechemokineswasbetween0.2and2whencomparingnotstimulatedwitha-toxin-stimulatedmacrophages.14Figure1Inductionofchemokine(C-X-Cmotif)ligand(CXCL)10followinga-toxinstimulationinhumanmacrophagesatthemRNAlevel.Figure2Inductionofchemokine(C-X-Cmotif)ligand(CXCL)10followinga-toxinstimulationinhumanmacrophagesattheproteinlevel.15Figure3Punchbiopsies(3mm)fromhealthyindividualswerelefteitherunstimulated(A)orstimulatedwitha-toxin(100ng/ml)(B)orIFN-c(100ng/ml)(C)for24hat37C.5-lmparaffinsectionswerestainedforCXCL10alongwithappropriateisotypeaswellasCD68.a-Toxin-stimulatedmacrophagescouldinducethemigrationofhumanCD4+TcellsviaCXCR316Bothinhibitors(anti-IP-10antibody&anti-CXCR3antibody)abrogatedthemigrationinducedbya-toxin-treatedmacrophagesupernatants.a-ToxinincellculturemediumwithoutmacrophagesdidnotdirectlyinducethemigrationofhumanCD4+lymphocytes.17Figure4Chemotacticactivityofa-toxin-stimulatedmacrophagesandinhibitionofchemokine(C-X-Cmotif)ligand(CXCL)10-inducedchemotaxisusinganeutralizinganti-CXCL10oranti-CXCR3Ab.Loweffectofa-toxinonCXCL10induction(Th1-relatedchemokine)inmacrophagesfrompatientswithAD18a-toxininducedsignificantlylowerlevelsofCXCL10expressionorsecretioninpatientswithADcomparedwithhealthycontrolsaswellaspsoriasisatalltimepointsanddosestested.19Figure5Macrophagesfrompatientswithatopicdermatitis(AD)andpsoriasis(PSO)aswellasfromhealthycontrols(HC)werelefteitherunstimulated(mediumcontrol)orstimulatedwitha-toxinasindicatedorwithIFN-c(10ng/ml)orTNF-a(20ng/ml),respectively,fortheindicatedperiodsoftime.Effectofa-toxinonMDCproduction(Th2-relatedchemokine)inmacrophagesfrompatientswithchronicinfammatoryskindiseases20MacrophagesfrompatientswithADaswellaspatientswithpsoriasisshowedanintrinsicallyhigherCCL22productioncomparedtohealthycontrolsbytrend.Onthewhole,a-toxindidnotregulateCCL22secretion.21Thankyou