嗜神經(jīng)病毒躲避宿主先天性免疫反應(yīng)的機(jī)制和應(yīng)用專家講座

嗜神經(jīng)病毒規(guī)避宿主先天性免疫反映旳機(jī)制及對(duì)策趙凌專家華中農(nóng)業(yè)大學(xué)10-15-2023第1頁Rabiesvirus180nmx75nm第2頁RobertHurt-USCRabiespathogenesisPatientsdieofcirculatoryinsufficiency,cardiacarrestandrespiratoryfailure.第3頁4RabiesinfectionandinnateimmunityWangetal.JournalofVirology,2023第4頁5Zhaoetal.JournalofVirology,2023重組狂犬病病毒構(gòu)建第5頁6ExpressionofMIP-1αattenuatedRABVpathogenicity,whileexpressionofRANTESorIP-10increasedRABVpathogencityZhaoetal.JournalofVirology,2023第6頁7ControlrHEPHEP-MIP1aHEP-RANTESHEP-IP10D3D6D9

HEstainingofmousebrainsZhaoetal.JournalofVirology,2023第7頁ExpressionofMIP-1α

enhancesVNAproductionandprotectionZhaoetal.JournalofVirology,2023第8頁Zhaoetal.JournalofVirology,2023外周過量體現(xiàn)MIP-1α

會(huì)吸引更多旳樹突狀細(xì)胞和B細(xì)胞第9頁第10頁AntigenicGroupVirusHostDiseaseI229ENL63humanhumanrespiratoryinfectionrespiratoryinfection,croupTGEVpigrespiratoryandentericinfectionCCVdogentericinfectionFECVcatentericinfectionFIPVcatrespiratory,enteric,hepatitisandneurologicalinfectionIIOC43humanRespiratoryandpossiblyentericinfectionSARS-CoVHKU1humanhumanRespiratoryRespiratoryMHVmouseRespiratory,enteric,neurologicinfectionHEVpigRespiratory,enteric,neurologicinfectionBCVcowEntericinfectionTCVturkeyRespiratoryandentericinfectionIIIIBVchickenRespiratoryandentericinfection,hepatitisCoronavirusescausediseasesinhumansanddomesticanimalsAdaptedfromHolmesandLai,FieldsVirology第11頁WhyMHV?MHVproducesabroadspectrumofdiseaseinthemouse-pneumonia(MHV-1)-hepatitis(MHV-A59)-encephalitis(MHV-A59/JHM)-demylination(MHV-A59)Itprovidesexcellentsmallanimalmodelsforhepatitis,forSARS,andformultiplesclerosis第12頁P(yáng)artI:

MHVns2interferestypeIinterferonresponses

第13頁Mutationofns2confers

attenuationofhepatitisbutnotCNSdisease

IC

500PFUIH

500PFURoth-Cross,J.K.etal,JVI,2023,83(8):3743-3753.brainliver第14頁Ns2isanorganspecificvirulencefactor

1a1b2a45a5bHESEMINMHV2Hphosphodiesterase;2predictedcatalyticHis-x-Thr/Sermotifs(Mazumdereta.,2023;Snijderetal.,2023)ns2(Mazumbderetal.,2023Snijderetal.,2023;Roth-Cross,2023)第15頁Mutationofns2confersattenuationofreplicationinmacrophagesandmicrogliabutnotinothercelltypes

Zhao,L.etal,JVI,2023.Oct;85(19):10058-10068.

第16頁MOI1MOI0.01

ns2mutantsrecovertheabilitytoreplicateefficientlyinmacrophagesandmicrogliafromIFNARknockoutmiceMOI1Zhao,L.etal,JVI,2023.Oct;85(19):10058-10068.

第17頁Type1interferoninductionandsignalingpathwaysIFN-IFNTYK2JAK1PSTAT1P

STAT1orSTAT2IFNAR1IFNAR2ISREIRF9PSTAT1PSTAT2ISGF3PSTAT2IRF-9ISGs:OAS,MxA,ISG15,ISG54,MDA5TLR3TLR7/8TLR9dsRNACpGssRNATIRTBK-1IKK-IRF-3IFN-NF-BATF-2RIG-ICBPPMDA5TIRTIRTRIFMyD88MyD88ISREIFN-第18頁L2AstroBMMIFNAR-/-BMMns2mutantsarenotdefectiveininductionofIFN-α/

mRNABothwtRA59andns2mutantsinduceminimalamountsofIFN-α,βmRNAinL2cellsandastrocyteswtRA59andns2mutantsinducesimilarlevelsofIFN-α,βmRNAinBMMfrombothB6andIFNAR-/-miceZhao,L.etal,JVI,2023.Oct;85(19):10058-10068.

第19頁BMMBMMMicroglians2mutantsaremoresensitivetotheantiviraleffectsofIFN-α/

thanwtA59inmacrophagesandmicrogliabutnotinothercelltypesL2Astro第20頁ViralreplicationandIFNsensitivityinthehepatocytesNoIFNIFN第21頁InvivomacrophagedepletionLiposomes,encapsulatingtheClodronatemolecules(squares),areingestedbymacrophagesviaendocytosis.Afterfusionwithlysosomes(L)containingphospholipases(arrowheads),thelatterdisruptthebilayersoftheliposomes.

Themoreconcentricbilayersaredisrupted,thegreateristheClodronatereleasewithinthecell.ThecellsarekilledbyClodronatethroughapoptosis.第22頁LiposomeClodronateA59ns2-H126RA59ns2-H126RPBSLiposomeClodronateH&EstainingNproteinstainingns2mutantsreplicateandinducehepatitisinmacrophagedepletedmice500foldvs10fold第23頁

Model:Kupffercellsprovideabarriertotheliverparenchymatoviruses=rA59LSECKCHepatocytesinusoidparenchyma=ns2-H126A第24頁P(yáng)artI:conclusionsns2isanorganspecificvirulencefactorandantagonizesIFNsignalingns2isrequiredforreplicationinmacrophages;depletionofmacrophagesinvivopromotesns2mutantvirusreplication

wesuggestthatMHVhastoreplicateinKupffercellsintheliversinusoidsinordertoreachtheliverparenchymaandinducehepatitis

第25頁Type1interferoninductionandsignalingpathwaysIFN-IFNTYK2JAK1PSTAT1P

STAT1orSTAT2IFNAR1IFNAR2ISREIRF9PSTAT1PSTAT2ISGF3PSTAT2IRF-9ISGs:OAS,MxA,ISG15,ISG54,MDA5TLR3TLR7/8TLR9dsRNACpGssRNATIRTBK-1IKK-IRF-3IFN-NF-BATF-2RIG-ICBPPMDA5TIRTIRTRIFMyD88MyD88ISREIFN-第26頁pCAGGS-IFNNS2a-IFNSV5V-IFNNDV-bioassayinVerocellspCAGGS-noIFNTansfectionofpCAGGSorpCAGGS-ns2orpCAGGS-SV5VinVerocellsTreatmentwithorw/o1000U/mlfor16hInfectionofNDV-GFP24h12hGFP第27頁ISGscreeninginKOBMMISG15IFIT1IFIT2PKRRNaseL第28頁P(yáng)artII:

MHVns2antagonizesOAS-RNaseLpathway第29頁InterferonsignalingmodelViraldsRNAMDA5*,RIG-I*IFNAntiviralISGsOAS*2-5ARNaseLCellularandviralRNASmallRNAs2’-PDEns2?MainpathwayOAS-RNaseLpathwayOAS=2’,5’-oliogoadenylatesynthetase2’-PDE=2’phosphodiesterase2-5A=2’,5’oligoadenylate第30頁WildtypeA59replicationinBMMwasnotaffectedbytheOAS-RNaseLsystem第31頁Defectivereplicationofns2mutantisrestoredinRNaseL-/-BMMButnotinPKR-/-BMMZhao,Letal.CellHost&Microbe,2023,(11)607–616.第32頁ns2expressionin293TcellspCAGGS-ns2pCAGGSpCAGGS-ns2-H126RZhao,Letal.CellHost&Microbe,2023,(11)607–616.第33頁ns2preventsrRNAcleavageand2-5AproductioninBMMZhao,Letal.CellHost&Microbe,2023,(11)607–616.第34頁Overexpressionofns2in293TcellspreventsrRNAcleavageand2-5AproductioninducedbypolyI:CZhao,Letal.CellHost&Microbe,2023,(11)607–616.第35頁Ns2cl