嗜神經病毒躲避宿主先天性免疫反應的機制及對策

1Rabiesvirus180nmx75nm2RobertHurt-USCRabiespathogenesisPatientsdieofcirculatoryinsufficiency,cardiacarrestandrespiratoryfailure.34RabiesinfectionandinnateimmunityWangetal.JournalofVirology,200545Zhaoetal.JournalofVirology,2009重組狂犬病病毒構建56ExpressionofMIP-1αattenuatedRABVpathogenicity,whileexpressionofRANTESorIP-10increasedRABVpathogencityZhaoetal.JournalofVirology,200967ControlrHEPHEP-MIP1aHEP-RANTESHEP-IP10D3D6D9

HEstainingofmousebrainsZhaoetal.JournalofVirology,20097ExpressionofMIP-1α

enhancesVNAproductionandprotectionZhaoetal.JournalofVirology,20108Zhaoetal.JournalofVirology,2010外周過量表達MIP-1α

會吸引更多的樹突狀細胞和B細胞910AntigenicGroupVirusHostDiseaseI229ENL63humanhumanrespiratoryinfectionrespiratoryinfection,croupTGEVpigrespiratoryandentericinfectionCCVdogentericinfectionFECVcatentericinfectionFIPVcatrespiratory,enteric,hepatitisandneurologicalinfectionIIOC43humanRespiratoryandpossiblyentericinfectionSARS-CoVHKU1humanhumanRespiratoryRespiratoryMHVmouseRespiratory,enteric,neurologicinfectionHEVpigRespiratory,enteric,neurologicinfectionBCVcowEntericinfectionTCVturkeyRespiratoryandentericinfectionIIIIBVchickenRespiratoryandentericinfection,hepatitisCoronavirusescausediseasesinhumansanddomesticanimalsAdaptedfromHolmesandLai,FieldsVirology11WhyMHV?MHVproducesabroadspectrumofdiseaseinthemouse-pneumonia(MHV-1)-hepatitis(MHV-A59)-encephalitis(MHV-A59/JHM)-demylination(MHV-A59)Itprovidesexcellentsmallanimalmodelsforhepatitis,forSARS,andformultiplesclerosis12PartI:

MHVns2interferestypeIinterferonresponses

13Mutationofns2confers

attenuationofhepatitisbutnotCNSdisease

IC

500PFUIH

500PFURoth-Cross,J.K.etal,JVI,2009,83(8):3743-3753.brainliver14Ns2isanorganspecificvirulencefactor

1a1b2a45a5bHESEMINMHV2Hphosphodiesterase;2predictedcatalyticHis-x-Thr/Sermotifs(Mazumdereta.,2002;Snijderetal.,2003)ns2(Mazumbderetal.,2002Snijderetal.,2003;Roth-Cross,2009)15Mutationofns2confersattenuationofreplicationinmacrophagesandmicrogliabutnotinothercelltypes

Zhao,L.etal,JVI,2011.Oct;85(19):10058-10068.

16MOI1MOI0.01

ns2mutantsrecovertheabilitytoreplicateefficientlyinmacrophagesandmicrogliafromIFNARknockoutmiceMOI1Zhao,L.etal,JVI,2011.Oct;85(19):10058-10068.

17Type1interferoninductionandsignalingpathwaysIFN-IFNTYK2JAK1STAT1

STAT1orSTAT2IFNAR1IFNAR2ISREIRF9STAT1STAT2ISGF3STAT2IRF-9ISGs:OAS,MxA,ISG15,ISG54,MDA5TLR3TLR7/8TLR9dsRNACpGssRNATIRTBK-1IKK-IRF-3IFN-NF-BATF-2RIG-ICBPPMDA5TIRTIRTRIFMyD88MyD88ISREIFN-18L2AstroBMMIFNAR-/-BMMns2mutantsarenotdefectiveininductionofIFN-α/

mRNABothwtRA59andns2mutantsinduceminimalamountsofIFN-α,βmRNAinL2cellsandastrocyteswtRA59andns2mutantsinducesimilarlevelsofIFN-α,βmRNAinBMMfrombothB6andIFNAR-/-miceZhao,L.etal,JVI,2011.Oct;85(19):10058-10068.

19BMMBMMMicroglians2mutantsaremoresensitivetotheantiviraleffectsofIFN-α/

thanwtA59inmacrophagesandmicrogliabutnotinothercelltypesL2Astro20ViralreplicationandIFNsensitivityinthehepatocytesNoIFNIFN21InvivomacrophagedepletionLiposomes,encapsulatingtheClodronatemolecules(squares),areingestedbymacrophagesviaendocytosis.Afterfusionwithlysosomes(L)containingphospholipases(arrowheads),thelatterdisruptthebilayersoftheliposomes.

Themoreconcentricbilayersaredisrupted,thegreateristheClodronatereleasewithinthecell.ThecellsarekilledbyClodronatethroughapoptosis.22LiposomeClodronateA59ns2-H126RA59ns2-H126RPBSLiposomeClodronateH&EstainingNproteinstainingns2mutantsreplicateandinducehepatitisinmacrophagedepletedmice500foldvs10fold23

Model:Kupffercellsprovideabarriertotheliverparenchymatoviruses=rA59LSECKCHepatocytesinusoidparenchyma=ns2-H126A24PartI:conclusionsns2isanorganspecificvirulencefactorandantagonizesIFNsignalingns2isrequiredforreplicationinmacrophages;depletionofmacrophagesinvivopromotesns2mutantvirusreplication

wesuggestthatMHVhastoreplicateinKupffercellsintheliversinusoidsinordertoreachtheliverparenchymaandinducehepatitis

25Type1interferoninductionandsignalingpathwaysIFN-IFNTYK2JAK1STAT1

STAT1orSTAT2IFNAR1IFNAR2ISREIRF9STAT1STAT2ISGF3STAT2IRF-9ISGs:OAS,MxA,ISG15,ISG54,MDA5TLR3TLR7/8TLR9dsRNACpGssRNATIRTBK-1IKK-IRF-3IFN-NF-BATF-2RIG-ICBPPMDA5TIRTIRTRIFMyD88MyD88ISREIFN-26pCAGGS-IFNNS2a-IFNSV5V-IFNNDV-bioassayinVerocellspCAGGS-noIFNTansfectionofpCAGGSorpCAGGS-ns2orpCAGGS-SV5VinVerocellsTreatmentwithorw/o1000U/mlfor16hInfectionofNDV-GFP24h12hGFP27ISGscreeninginKOBMMISG15IFIT1IFIT2PKRRNaseL28PartII:

MHVns2antagonizesOAS-RNaseLpathway29InterferonsignalingmodelViraldsRNAMDA5*,RIG-I*IFNAntiviralISGsOAS*2-5ARNaseLCellularandviralRNASmallRNAs2’-PDEns2?MainpathwayOAS-RNaseLpathwayOAS=2’,5’-oliogoadenylatesynthetase2’-PDE=2’phosphodiesterase2-5A=2’,5’oligoadenylate30WildtypeA59replicationinBMMwasnotaffectedbytheOAS-RNaseLsystem31Defectivereplicationofns2mutantisrestoredinRNaseL-/-BMMButnotinPKR-/-BMMZhao,Letal.CellHost&Microbe,2012,(11)607–616.32ns2expressionin293TcellspCAGGS-ns2pCAGGSpCAGGS-ns2-H126RZhao,Letal.CellHost&Microbe,2012,(11)607–616.33ns2preventsrRNAcleavageand2-5AproductioninBMMZhao,Letal.CellHost&Microbe,2012,(11)607–616.34Overexpressionofns2in293TcellspreventsrRNAcleavageand2-5AproductioninducedbypolyI:CZhao,Letal.CellHost&Microbe,2012,(11)607–616.35Ns2cleaves2-5AintoATPandAMPZhao,Letal.CellHost&