Dynasore, a Dynamin Inhibitor, Inhibits Trypanosoma cruzi Entry into Peritoneal Macrophages 英文参考文献.docVIP

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Dynasore, a Dynamin Inhibitor, Inhibits Trypanosoma cruzi Entry into Peritoneal Macrophages 英文参考文献.doc

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Dynasore, a Dynamin Inhibitor, Inhibits Trypanosoma cruzi Entry into Peritoneal Macrophages 英文参考文献

Dynasore,aDynaminInhibitor,InhibitsTrypanosoma cruziEntryintoPeritonealMacrophages EmileS.Barrias,LissaC.Reignault,WanderleyDeSouza,TeciaM.U.Carvalho* Laborato′riodeUltraestruturaCelularHerthaMeyer,InstitutodeBiof?′sicaCarlosChagasFilho,UniversidadeFederaldoRiodeJaneiro,CCS-BlocoG,IlhadoFunda?o,Riode Janeiro,Brasil Abstract Background: Trypanosoma cruzi is an intracellular parasite that, like some other intracellular pathogens, targets specific proteinsofthehostcellvesiculartransportmachinery,leadingtoamodulationofhostcellprocessesthatresultsinthe generationofuniquephagosomes.Inmammaliancells,severalmoleculeshavebeenidentifiedthatselectivelyregulatethe formation ofendocytic transport vesicles andthefusion of suchvesicles withappropriate acceptor membranes. Among these, the GTPase dynamin plays an important role in clathrin-mediated endocytosis, and it was recently found that dynamincanparticipateinaphagocyticprocess. Methodology/PrincipalFindings:WeusedacompoundcalleddynasorethathastheabilitytoblocktheGTPaseactivityof dynamin.Dynasoreactsasapotentinhibitorofendocyticpathwaysbyblockingcoatedvesicleformationwithinsecondsof its addition. Here, we investigated whether dynamin is involved in the entry process of T. cruzi in phagocytic and non- phagocyticcellsbyusingdynasore.Inthisaim,peritonealmacrophages andLLC-MK2cellsweretreatedwithincreasing concentrations of dynasorebefore interactionwith trypomastigotes, amastigotes orepimastigotes. Weobserved that,in both cell lines, the parasite internalization was drastically diminished (by greater than 90% in LLC-MK2 cells and 70% in peritonealmacrophages)whenweused100mMdynasore.TheT.cruziadhesionindex,however,wasunaffectedineither cellline.AnalyzingtheseinteractionsbyscanningelectronmicroscopyandcomparingperitonealmacrophagestoLLC-MK2 cellsrevealeddifferencesinthestageatwhichcellentrywasblocked.InLLC-MK2cells,thisblockadeisobservedearlier thanitisinperitonealmacrophages.InLLC-MK2cells,theparasiteswereonlyassociatedwithc