Activation of the Innate Immune Response against DENV in Normal Non-Transformed Human Fibroblasts 英文参考文献.docVIP

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Activation of the Innate Immune Response against DENV in Normal Non-Transformed Human Fibroblasts 英文参考文献.doc

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Activation of the Innate Immune Response against DENV in Normal Non-Transformed Human Fibroblasts 英文参考文献

ActivationoftheInnateImmuneResponseagainstDENV inNormalNon-TransformedHumanFibroblasts Jose′ Bustos-Arriaga1,Jazm?′nGarc?′a-Machorro1,Moise′sLeo′n-Jua′rez1,JulioGarc?′a-Cordero1 ,Leopoldo Santos-Argumedo1,LeopoldoFlores-Romo2,A.Rene′ Me′ndez-Cruz3,FranciscoJ.Jua′rez-Delgado4, LeticiaCedillo-Barro′n1* 1DepartamentodeBiomedicinaMolecularCentrodeInvestigacio′nydeEstudiosAvanzados,Me′xicoDistritoFederal,Mexico,2DepartamentodeBiolog?′aCelularCentro de Investigacio′n y de Estudios Avanzados, Me′xico Distrito Federal, Mexico, 3Laboratorio de Inmunolog?′a UMF de la FES Iztacala Universidad Auto′noma de Me′xico, TlalnepantlaEstadodeMe′xico,Mexico,4DepartamentodeCirugia,HospitalSantaMariaTicoma′n, Me′xicoDistritoFederal,Mexico Abstract Background:WhenmosquitoesinfectedwithDENVarefeeding,theproboscismusttraversetheepidermisseveraltimes (‘‘probing’’)beforereachingabloodvesselinthedermis.Duringthisprocess,thesalivaryglandsreleasethevirus,whichis likelytointeractfirstwithcellsofthevariousepidermalanddermallayers,cellswhichcouldbephysiologicallyrelevantto DENVinfectionandreplicationinhumans.However,importantquestionsarewhethermoreabundantnon-hematopoietic cells such as fibroblasts become infected, and whether they play any role in antiviral innate immunity in the very early stagesofinfection,oreveniftheymightbeusedbyDENVasprimaryreplicationcells. Methodology/PrincipalFindings:Fibroblastsfreshlyreleasedfromhealthyskinandinfected12hoursaftertheirisolation show a positive signal for DENV. In addition, when primary skin fibroblast cultures were established and subsequently infected, we showed DENV-2 antigen-positive intracellular signal at 24hours and 48hours post-infection. Moreover, the fibroblasts showed productive infection in a conventional plaque assay. The skin fibroblasts infected with DENV-2 underwentpotentsignalingthroughbothTLR3andRIG-1,butnotMda5,triggeringup-regulationofIFNb,TNFa,defensin5 (HB5)andbdefensin2(HbD2).Inaddition,DENVinfectedfibroblastsshowedincrease