简介：Usingtwo-colourflowcytometry＞200antibodiessubmittedtothe8thInternationalWorkshopofHumanLeukocyteDifferentiationAntigens(HLDA8)havebeenanalyzedfortheirreactivitywithrestingandactivatedCD203c+basophils.Fourantibodieseithernon-reactiveorweaklyreactivewithrestingbasophilsexhibitedanincreasedreactivitywithbasophilsactivatedbyanti-IgE-mediatedcross-linkingofthehighaffinityIgEreceptor(FcεRI).TheseincludeantibodiesagainstCD164(WS-80160,cloneN6B6andWS-80162,clone67D2),aswellastworeagentswithpreviouslyunknownspecificitiesthatwereidentifiedasCD13(WS-80274,cloneA8)andCD107a(WS-80280,cloneE63-880).Theactivationpatternsfollowedeitherthe'CD203c-like'or'CD63-like'activationprofile.TheCD203cprofileischaracterizedbyarapidandsignificantupregulation(ofCD13,CD164,andCD203c),reachingmaximumlevelsafter5-15minofstimulation.ThePhosphoinositide-3-kinase(PI3K)-specificinhibitorWortmannininhibitedtheupregulationofthesemarkerswhereas12-O-tetradecanoyl-phorbol-13-acetate(TPA)inducedarapidandFcεRI-independentupregulationwithin1-2min.IntheCD63profile,maximumupregulation(ofCD63andCD107a)wasdetectedonlyafter20-40min,andupregulationbyTPAreachedmaximumlevelsafter60min.Insummary,ourdataidentifyCD13,CD107a,andCD164asnovelbasophil-activationantigens.Basedontimekineticsofupregulation,wehypothesizethatmoleculesofthe'CD203cgroup'andthe'CD63group'arelinkedtotwodifferentmechanismsofbasophilactivation.

简介：IL-16isaligandandchemotacticfactorforCD4+Tcells.IL-16inhibitstheCD3mediatedlymphocyteactivationandproliferation.TheeffectsofIL-16onthetargetcellsaredependentonthecelltype,thepresenceofco-activatorsetc.TounderstandtheregulationfunctionandmechanismofIL-16ontargetcells,weuseda130a.a.recombinantIL-16tostudyitseffectsonthegrowthofJurkatTleukemiacellsinvitro.WefoundthattherIL-16stimulatedtheproliferationofJurkatcellsatlowdose(10^-9M),butinhibitedthegrowthofthecellsathigherconcentration(10^-5M).Resultsshowedthat10^-5MofrIL-16treatmentinducedanenhancedapoptosisinJurkatcells.ThetreatmentblockedtheexpressionofFasL,butup-regulatedthec-mycandBidexpressioninthecells.Pre-treatmentofPKCinhibitororMEK1inhibitormarkedlyincreasedordecreasedtherIL-16inducedgrowth-inhibitingeffectsonJurkatcells,respectively.TheresultssuggestedthattherIL-16mightbearegulatorforthegrowthorapoptosisofJurkatcellsatadose-dependentmanner.Thegrowth-inhibitingeffectsofrIL-16mightbeFas/FasLindependent,but,associatedwiththeactivationofPKC,up-regulatedexpressionofc-MycandBid,andtheparticipationoftheERKsignalpathwayinJurkatcells.

简介：BAC-TO-BAC杆状病毒表达系统是一种快速、高效、便捷的表达系统.将人可溶性CD14(sCD14)基因克隆入pFASTBAC1转移质粒中,重组质粒转化DH10BAC感受态细胞,目的基因通过同源重组插入BacmidDNA中,后者转染sf21昆虫细胞获得重组杆状病毒.利用重组蛋白C-末端的6×His@Tag,经TALON金属螯合色谱将重组病毒感染昆虫细胞获得的无血清培养上清--步纯化得到重组蛋白,计算表明从1L培养基中可纯化到约8mg纯度大于95%的重组sCD14蛋白,免疫印迹结果表明重组蛋白具有与抗6×His单抗和抗CD14单抗结合的抗原性.疑胶迁移实验和细胞活性实验表明重组sCD14蛋白能在体外与LPS结合,并能增强LPS诱导THP-1细胞产生细胞毒性因子.

简介：<正>Uponactivation,naiveT-helpercellscandifferentiateintotwomajordistinctsubsets,Thelper1(Th1)andThelper2(Th2),asdefinedbytheireffectorfunctionsandcytokinesecretionpatterns.CytokinemilieuandcostimulatorymoleculeshavebeenshowntoplayanessentialroleindeterminingThelperdifferentiation.However,itisstillunclearhowtheeffectsofsignalsofco-stimulatorymoleculesandcytokinesareexertedduringThelperdifferentiation.Weshowevidencesuggestingthatwhilecytokinesignalsinitiatedifferentiationprogram,theselectiveactionofdeatheffectorsdeterminestheendpointbalanceofdifferenti-