Alternative Oxidase Mediates Pathogen Resistance in Paracoccidioides brasiliensis Infection 英文参考文献.docVIP

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Alternative Oxidase Mediates Pathogen Resistance in Paracoccidioides brasiliensis Infection 英文参考文献.doc

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Alternative Oxidase Mediates Pathogen Resistance in Paracoccidioides brasiliensis Infection 英文参考文献

AlternativeOxidaseMediatesPathogenResistancein ParacoccidioidesbrasiliensisInfection OrvilleHerna′ndezRuiz1,2,3*,AngelGonzalez4,5,AgostinhoJ.Almeida2,DianaTamayo2,AnaMaria Garcia2,AngelaRestrepo4,JuanG.McEwen2,6 1InstitutodeBiolog?′a,UniversidaddeAntioquia,Medell?′n,Colombia,2CellularandMolecularBiologyUnit,Corporacio′nparaInvestigacionesBiolo′gicas(CIB),Medell?′n, Colombia,3Facultad deCiencias delaSalud,Institucio′nUniversitaria ColegioMayordeAntioquia,Medell?′n,Colombia,4MedicalandExperimentalMycology Group, Corporacio′nparaInvestigacionesBiolo′gicas(CIB),Medell?′n,Colombia,5EscueladeMicrobiologia,UniversidaddeAntioquia,Medell?′n,Colombia,6FacultaddeMedicina, UniversidaddeAntioquia,Medell?′n,Colombia Abstract Background:Paracoccidioidesbrasiliensisisahumanthermaldimorphicpathogenicfungus.SurvivalofP.brasiliensisinside thehostdependsontheadaptationofthisfungalpathogentodifferentconditions,namelyoxidativestressimposedby immunecells. Aims and Methodology: In this study, we evaluated the role of alternative oxidase (AOX), an enzyme involved in the intracellular redox balancing, during host-P. brasiliensis interaction. We generated a mitotically stable P. brasiliensis AOX (PbAOX)antisenseRNA(aRNA)strainwitha70%reductioningeneexpression.WeevaluatedtherelevanceofPbAOXduring interaction of conidia and yeast cells with IFN-c activated alveolar macrophages and in a mouse model of infection. Additionally,wedeterminedthefungalcell’sviabilityandPbAOXinthepresenceofH2O2. Results: InteractionwithIFN-cactivatedalveolarmacrophagesinducedhigherlevelsof PbAOXgeneexpressioninPbWt conidia than PbWt yeast cells. PbAOX-aRNA conidia and yeast cells had decreased viability after interaction with macrophages. Moreover, in a mouse model of infection, we showed that absence of wild-type levels of PbAOX in P. brasiliensisresultsinareducedfungalburdeninlungsatweeks8and24post-challengeandanincreasedsurvivalrate.In thepresenceofH2O2,weobservedthatPbWtyeastcellsincreasedPbAOXexpres