嗜神经病毒躲避宿主先天性免疫反应的机制和应用培训课件.ppt

1、嗜神经病毒躲避宿主嗜神经病毒躲避宿主先天性免疫反应的机先天性免疫反应的机制和应用制和应用Rabies virus180nm x 75nm嗜神经病毒躲避宿主先天性免疫反应的机制和应用2Robert Hurt-USCRabies pathogenesis嗜神经病毒躲避宿主先天性免疫反应的机制和应用34Rabies infection and innate immunityWang et al.Journal of Virology,2005嗜神经病毒躲避宿主先天性免疫反应的机制和应用45Zhao et al.Journal of Virology,2009重组狂犬病病毒构建重组狂犬病病毒构建嗜神经

2、病毒躲避宿主先天性免疫反应的机制和应用56Expression of MIP-1MIP-1 attenuated RABV pathogenicity,while expression of RANTES or IP-10 increased RABV pathogencityZhao et al.Journal of Virology,2009嗜神经病毒躲避宿主先天性免疫反应的机制和应用67Control rHEP HEP-MIP1a a HEP-RANTES HEP-IP10D3D6D9 HE staining of mouse brainsZhao et al.Journal of Vi

3、rology,2009嗜神经病毒躲避宿主先天性免疫反应的机制和应用7Expression of MIP-1 MIP-1 enhances VNA production and protectionZhao et al.Journal of Virology,2010嗜神经病毒躲避宿主先天性免疫反应的机制和应用8Zhao et al.Journal of Virology,2010外周过量表达外周过量表达MIP-1会会吸引更多的树突状细胞和吸引更多的树突状细胞和B细胞细胞嗜神经病毒躲避宿主先天性免疫反应的机制和应用9嗜神经病毒躲避宿主先天性免疫反应的机制和应用10 Antigenic Group

4、VirusHostDiseaseI229ENL63humanhumanrespiratory infectionrespiratory infection,croupTGEVpigrespiratory and enteric infectionCCVdogenteric infectionFECVcatenteric infectionFIPVcatrespiratory,enteric,hepatitis and neurological infectionIIOC43humanRespiratory and possibly enteric infectionSARS-CoVHKU1hu

5、manhumanRespiratoryRespiratoryMHVmouseRespiratory,enteric,neurologic infectionHEVpigRespiratory,enteric,neurologic infectionBCVcowEnteric infectionTCVturkeyRespiratory and enteric infection IIIIBVchickenRespiratory and enteric infection,hepatitisCoronaviruses cause diseases in humans and domestic an

6、imalsAdapted from Holmes and Lai,Fields Virology嗜神经病毒躲避宿主先天性免疫反应的机制和应用11Why MHV?MHV produces a broad spectrum of disease in the mouse -pneumonia(MHV-1)-hepatitis(MHV-A59)-encephalitis(MHV-A59/JHM)-demylination(MHV-A59)It provides excellent small animal models for hepatitis,for SARS,and for multiple

7、sclerosis嗜神经病毒躲避宿主先天性免疫反应的机制和应用12Part I:MHV ns2 interferes type I interferon responses 嗜神经病毒躲避宿主先天性免疫反应的机制和应用13Mutation of ns2 confers attenuation of hepatitis but not CNS disease IC 500PFUIH 500PFU Roth-Cross,J.K.et al,JVI,2009,83(8):3743-3753.brainliver嗜神经病毒躲避宿主先天性免疫反应的机制和应用14Ns2 is an organ speci

8、fic virulence factor 1a1b2a4 5a5bHESEMINMHV 2H phosphodiesterase;2 predicted catalytic His-x-Thr/Ser motifs(Mazumder et a.,2002;Snijder et al.,2003)ns2(Mazumbder et al.,2002Snijder et al.,2003;Roth-Cross,2009)嗜神经病毒躲避宿主先天性免疫反应的机制和应用15Mutation of ns2 confers attenuation of replication in macrophages a

9、nd microglia but not in other cell types Zhao,L.et al,JVI,2011.Oct;85(19):10058-10068.嗜神经病毒躲避宿主先天性免疫反应的机制和应用16MOI 1MOI 0.01 ns2 mutants recover the ability to replicate efficiently in macrophages and microglia from IFNAR knockout miceMOI 1 Zhao,L.et al,JVI,2011.Oct;85(19):10058-10068.嗜神经病毒躲避宿主先天性免疫反

10、应的机制和应用17Type 1 interferon induction and signaling pathwaysIFN-IFNTYK2JAK1PSTAT1P STAT1 orSTAT2IFNAR1IFNAR2ISREIRF9PSTAT1PSTAT2 ISGF3PSTAT2IRF-9ISGs:OAS,MxA,ISG15,ISG54,MDA5TLR3TLR7/8TLR9dsRNACpGssRNATIRTBK-1IKK-IRF-3IFN-NF-BATF-2RIG-ICBPPMDA5TIRTIRTRIFMyD88MyD88ISREIFN-嗜神经病毒躲避宿主先天性免疫反应的机制和应用18L2Ast

11、roBMMIFNAR-/-BMMns2 mutants are not defective in induction of IFN-/mRNABoth wt RA59 and ns2 mutants induce minimal amounts of IFN-,mRNA in L2 cells and astrocytes wt RA59 and ns2 mutants induce similar levels of IFN-,mRNA in BMM from both B6 and IFNAR-/-mice Zhao,L.et al,JVI,2011.Oct;85(19):10058-10

12、068.嗜神经病毒躲避宿主先天性免疫反应的机制和应用19BMMBMMMicroglians2 mutants are more sensitive to the antiviral effects of IFN-IFN-/than wt A59 in macrophages and microgliabut not in other cell typesL2Astro嗜神经病毒躲避宿主先天性免疫反应的机制和应用20Viral replication and IFN sensitivity in the hepatocytesNo IFNIFN嗜神经病毒躲避宿主先天性免疫反应的机制和应用21In

13、 vivo macrophage depletionLiposomes,encapsulating the Clodronate molecules(squares),are ingested by macrophages via endocytosis.After fusion with lysosomes(L)containing phospholipases(arrowheads),the latter disrupt the bilayers of the liposomes.The more concentric bilayers are disrupted,the greater

14、is the Clodronate release within the cell.The cells are killed by Clodronate through apoptosis.嗜神经病毒躲避宿主先天性免疫反应的机制和应用22Liposome ClodronateA59ns2-H126RA59ns2-H126RPBSLiposome ClodronateH&E stainingN protein stainingns2 mutants replicate and induce hepatitis in macrophage depleted mice500 fold vs 10 f

15、old嗜神经病毒躲避宿主先天性免疫反应的机制和应用23Part I:conclusions ns2 is an organ specific virulence factor and antagonizes IFN signaling ns2 is required for replication in macrophages;depletion of macrophages in vivo promotes ns2 mutant virus replication we suggest that MHV has to replicate in Kupffer cells in the liv

16、er sinusoids in order to reach the liver parenchyma and induce hepatitis 嗜神经病毒躲避宿主先天性免疫反应的机制和应用25Type 1 interferon induction and signaling pathwaysIFN-IFNTYK2JAK1PSTAT1P STAT1 orSTAT2IFNAR1IFNAR2ISREIRF9PSTAT1PSTAT2 ISGF3PSTAT2IRF-9ISGs:OAS,MxA,ISG15,ISG54,MDA5TLR3TLR7/8TLR9dsRNACpGssRNATIRTBK-1IKK-

17、IRF-3IFN-NF-BATF-2RIG-ICBPPMDA5TIRTIRTRIFMyD88MyD88ISREIFN-嗜神经病毒躲避宿主先天性免疫反应的机制和应用26pCAGGS-IFNNS2a-IFNSV5V-IFNNDV-bioassay in Vero cellspCAGGS-no IFNTansfection of pCAGGSor pCAGGS-ns2 or pCAGGS-SV5Vin Vero cellsTreatment with or w/o1000U/ml for 16hInfection of NDV-GFP24h12hGFP 嗜神经病毒躲避宿主先天性免疫反应的机制和应用2

18、7ISG screening in KO BMMISG15IFIT1IFIT2PKRRNase L嗜神经病毒躲避宿主先天性免疫反应的机制和应用28Part II:MHV ns2 antagonizes OAS-RNase L pathway嗜神经病毒躲避宿主先天性免疫反应的机制和应用29Interferon signaling modelViral dsRNAMDA5*,RIG-I*IFNAntiviral ISGsOAS*2-5ARNase LCellular and viral RNASmall RNAs2-PDEns2?Main pathwayOAS-RNase L pathwayOAS

19、=2,5-oliogoadenylate synthetase2-PDE=2 phosphodiesterase2-5A=2,5oligoadenylate嗜神经病毒躲避宿主先天性免疫反应的机制和应用30Wild type A59 replication in BMM was not affected by the OAS-RNase L system嗜神经病毒躲避宿主先天性免疫反应的机制和应用31Defective replication of ns2 mutant is restored in RNase L-/-BMMBut not in PKR-/-BMMZhao,L et al.Ce

20、ll Host&Microbe,2012,(11)607616.嗜神经病毒躲避宿主先天性免疫反应的机制和应用32ns2 expression in 293T cellspCAGGS-ns2pCAGGSpCAGGS-ns2-H126RZhao,L et al.Cell Host&Microbe,2012,(11)607616.嗜神经病毒躲避宿主先天性免疫反应的机制和应用33ns2 prevents rRNA cleavage and 2-5A production in BMMZhao,L et al.Cell Host&Microbe,2012,(11)607616.嗜神经病毒躲避宿主先天性免

21、疫反应的机制和应用34Overexpression of ns2 in 293T cells prevents rRNA cleavage and 2-5A production induced by poly I:CZhao,L et al.Cell Host&Microbe,2012,(11)607616.嗜神经病毒躲避宿主先天性免疫反应的机制和应用35Ns2 cleaves 2-5A into ATP and AMPZhao,L et al.Cell Host&Microbe,2012,(11)607616.嗜神经病毒躲避宿主先天性免疫反应的机制和应用36Ns2 enhances vir

22、us replication in the liverZhao,L et al.Cell Host&Microbe,2012,(11)607616.嗜神经病毒躲避宿主先天性免疫反应的机制和应用37Ns2 facilitates the induction of viral hepatitisZhao,L et al.Cell Host&Microbe,2012,(11)607616.嗜神经病毒躲避宿主先天性免疫反应的机制和应用38l Mouse hepatitis virus ns2 protein inhibits the IFN-induced OAS-RNase L pathway l

23、ns2 reduces the intracellular level of 2,5-oligoadenylate(2-5A)l ns2 has a 2,5-phosphodiesterase to directly cleave 2-5A l ns2 mutant virus replication and hepatitis-induction are rescued in RNase L deficient micePart II:conclusions嗜神经病毒躲避宿主先天性免疫反应的机制和应用391.狂犬病病毒如何逃逸中枢神经系统先天性免疫反应？2.狂犬病病毒如何限制血脑屏障的打开？3.如何打开血脑屏障清除狂犬病病毒？研究方向研究方向嗜神经病毒躲避宿主先天性免疫反应的机制和应用4041谢 谢！嗜神经病毒躲避宿主先天性免疫反应的机制和应用41