Enterohemorrhagic Escherichia coli Specific Enterohemolysin Induced IL-1β in Human Macrophages and EHEC-Induced IL-1β Required Activation of NLRP3 Inflammasome 英文参考文献.docVIP

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Enterohemorrhagic Escherichia coli Specific Enterohemolysin Induced IL-1β in Human Macrophages and EHEC-Induced IL-1β Required Activation of NLRP3 Inflammasome 英文参考文献.doc

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Enterohemorrhagic Escherichia coli Specific Enterohemolysin Induced IL-1β in Human Macrophages and EHEC-Induced IL-1β Required Activation of NLRP3 Inflammasome 英文参考文献

EnterohemorrhagicEscherichiacoliSpecific EnterohemolysinInducedIL-1binHumanMacrophages andEHEC-InducedIL-1bRequiredActivationofNLRP3 Inflammasome XiaoaiZhang1,2.,YuliCheng1.,YanwenXiong1,ChangyunYe1,HanZheng1,HuiSun1,HongqingZhao1, ZhihongRen1*,JianguoXu1* 1NationalInstituteforCommunicableDiseaseControlandPrevention,ChineseCenterforDiseaseControlandPrevention,StateKeyLaboratoryforInfectiousDisease PreventionandControl,Changping,Beijing,China,2BeijingCenterforDiseasePreventionandControl,Beijing,China Abstract Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a major foodborne pathogen causing hemorrhagic colitis and hemolytic-uremic syndrome. The role of EHEC O157:H7-enterohemolysin (Ehx) in the pathogenesis of infections remains poorlydefined.Inthisstudy,weusedgenedeletionandcomplementmethodstoconfirmitsputativefunctions.Results demonstratedthat,inTHP-1cells,EHECO157:H7-Ehxisassociatedwithgreaterproductionofextracellularinterleukin(IL)-1b thanothercytokines.ThedataalsoshowedthatEHECO157:H7-EhxcontributedtocytotoxicityinTHP-1cells,causingthe release of lactate dehydrogenase (LDH). Although we observed a positive correlation between IL-1b production and cytotoxicityinTHP-1cellsinfectedwithdifferentEHECO157:H7strains,ourimmunoblotresultsshowedthatthemajorityof IL-1b in the supernatant was mature IL-1b and not the pro-IL-1b that can be released after cell death. However, EHEC O157:H7-Ehxhadnodetectableeffectonbiologicallyinactivepro-IL-1batthemRNAorproteinsynthesislevels.Neitherdid itaffecttheexpressionofapoptosis-associatedspeck-likeproteincontainingaCARD(ASC),caspase-1,orNOD-likereceptor family pyrin domain containing 3 (NLRP3). RNA interference experiments showed that EHEC O157:H7-induced IL-1b productionrequiredtheinvolvementofASC,caspase-1,andNLRP3expressioninTHP-1cells.Ourresultsdemonstratethat EhxplaysacrucialroleinEHECO157:H7-inducedIL-1bproductionanditscytotoxicitytoTHP-1cells.NLRP3inflammasome activationisalsoinvolvedinEHECO157:H7-stimulatedIL-1brelease.