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1、嗜神经病毒躲避宿主先天性免疫反应的机制及对策,赵 凌 教授10-15-2013,嗜神经病毒躲避宿主先天性免疫反应的机制及对策赵 凌 教,Rabies virus,180nm x 75nm,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,Rabies virus180nm x 75nm嗜神经病毒躲,Robert Hurt-USC,Rabies pathogenesis,Patients die of circulatory insufficiency, cardiac arrest and respiratory failure.,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,Ro
2、bert Hurt-USCRabies pathogen,4,Rabies infection and innate immunity,Wang 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及应用赵凌老师,4Rabies infection and innate i,5,Zhao et al. Journal of Virology, 2009,重组狂犬病病毒构建,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,5Zhao et al. Journal of Virolo,6,Expression of MIP-1 attenu
3、ated RABV pathogenicity, while expression of RANTES or IP-10 increased RABV pathogencity,Zhao 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及应用赵凌老师,6Expression of MIP-1 attenuat,7,Control rHEP HEP-MIP1a HEP-RANTES HEP-IP10,D3D6D9,HE staining of mouse brains,Zhao et al. Journal of Virology, 200
4、9,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,7Control rHEP,Expression of MIP-1 enhances VNA production and protection,Zhao 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及应用赵凌老师,Expression of MIP-1 enhances,Zhao et al. Journal of Virology, 2010,外周过量表达MIP-1会吸引更多的树突状细胞和B细胞,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,Zhao et al.
5、Journal of Virolog,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,Coronaviruses cause diseases in humans and domestic animals,Adapted 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及应用赵凌老师,Antigenic GroupVirusHostDisea,Why MHV?,MHV produces a broad spectrum of disease in the mouse -p
6、neumonia (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 sclerosis,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,Why MHV?MHV produces a broad s,Part I: MHV ns2 interferes type I interferon responses,Part
7、 I: MHV ns2 interferes t,Mutation of ns2 confers attenuation of hepatitis but not CNS disease,IC 500PFU,IH 500PFU,Roth-Cross, J.K. et al, JVI, 2009, 83(8):3743-3753.,brain,liver,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,Mutation of ns2 confers atten,Ns2 is an organ specific virulence factor,2H phosphodiesterase; 2
8、 predicted catalytic His-x-Thr/Ser motifs (Mazumder et a., 2002; Snijder et al.,2003),(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及应用赵凌老师,Ns2 is an organ specific virul,Mutation of ns2 confers attenuation of replication in macrophages and microglia but not in oth
9、er 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及应用赵凌老师,Mutation of ns2 confers attenu,MOI 1,MOI 0.01,ns2 mutants recover the ability to replicate efficiently in macrophages and microglia from IFNAR knockout mice,MOI 1,Zhao, L. et al, JVI, 2011.Oct; 85(19):10058-1
10、0068.,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,MOI 1MOI 0.01 ns2 mutants reco,Type 1 interferon induction and signaling pathways,IFN-,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,Type 1 interferon induction an,L2,Astro,BMM,IFNAR-/- BMM,ns2 mutants are not defective in induction of IFN-/ mRNA,Both wt RA59 and ns2 mutants induce min
11、imal 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):10058-10068.,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,L2AstroBMMIFNAR-/- BMMns2 muta,BMM,BMM,Microglia,ns2 mutants are more
12、sensitive to the antiviral effects of IFN-/ than wt A59 in macrophages and microglia,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,BMMBMMMicroglians2 mutants are,Viral replication and IFN sensitivity in the hepatocytes,No IFN,IFN,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,Viral replication and IFN sens,In vivo macrophage depletion,Li
13、posomes, 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 is the Clodronate
14、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及应用赵凌老师,In vivo macrophage depletionLi,ns2 mutants replicate and induce hepatitis in macrophage depleted mice,500 fold vs 10 fold,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,Liposome ClodronateA59ns2-H126,Model: Kupffer
15、 cells provide a barrier to the liver parenchyma to viruses,=rA59,LSEC,KC,Hepatocyte,sinusoid,parenchyma,=ns2-H126A,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,Model: =rA59LSECKCHepatocytes,Part I: conclusions,ns2 is an organ specific virulence factor and antagonizes IFN signalingns2 is required for replication in m
16、acrophages; depletion of macrophages in vivo promotes ns2 mutant virus replicationwe suggest that MHV has to replicate in Kupffer cells in the liver 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及应用赵凌老师,Part I: conclusions嗜神经病毒躲避宿主先天,Type 1 interferon induc
17、tion and signaling pathways,IFN-,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,Type 1 interferon induction an,pCAGGS-IFN,NS2a-IFN,SV5V-IFN,NDV-bioassay in Vero cells,pCAGGS-no IFN,Tansfection of pCAGGSor pCAGGS-ns2 or pCAGGS-SV5Vin Vero cells,Treatment with or w/o1000U/ml for 16h,Infection of NDV-GFP,24h,12h,GFP,嗜神经病毒
18、躲避宿主先天性免疫反应的机制及应用赵凌老师,pCAGGS-IFNNS2a-IFNSV5V-IFNNDV-,ISG screening in KO BMM,ISG15,IFIT1,IFIT2,PKR,RNase L,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,ISG screening in KO BMMISG15IF,Part II: MHV ns2 antagonizes OAS-RNase L pathway,Part II: MHV ns2 antagonizes,Interferon signaling model,Viral dsRNA,MDA5*, RIG-I*,IFN,
19、Antiviral ISGs,OAS*,2-5A,RNase L,Cellular and viral RNA,Small RNAs,2-PDE,ns2?,Main pathway,OAS-RNase L pathway,OAS= 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及应用赵凌老师,Interferon signaling modelVira,Wild type A59 replication in BMM was not affect
20、ed 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及应用赵凌老师,Wild type A59 replication in B,Defective replication of ns2 mutant is restored in RNase L-/- BMM,But not in PKR-/- BMM,Zhao, 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及应用赵凌老师,Defective replication of ns2 m,ns2 expressi
21、on in 293T cells,pCAGGS-ns2,pCAGGS,pCAGGS-ns2-H126R,Zhao, 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及应用赵凌老师,ns2 expression in 293T cellspC,ns2 prevents rRNA cleavage and 2-5A production in BMM,Zhao, 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及应用赵凌老师
22、,ns2 prevents rRNA cleavage and,Overexpression of ns2 in 293T cells prevents rRNA cleavage and 2-5A production induced by poly I:C,Zhao, 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及应用赵凌老师,Overexpression of ns2 in 293T,Ns2 cleaves 2-5A into ATP and AMP,Zhao, L et al. Cell Hos
23、t & Microbe, 2012, (11) 607616.,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,Ns2 cleaves 2-5A into ATP and,Ns2 enhances virus replication in the liver,Zhao, 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及应用赵凌老师,Ns2 enhances virus replication,Ns2 facilitates the induction of viral hepatitis,Zhao,
24、 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及应用赵凌老师,Ns2 facilitates the induction,Mouse hepatitis virus ns2 protein inhibits the IFN-induced OAS-RNase L pathway ns2 reduces the intracellular level of 2,5-oligoadenylate (2-5A) ns2 has a 2,5-phosphodiesterase to directly cleav
25、e 2-5A ns2 mutant virus replication and hepatitis-induction are rescued in RNase L deficient mice,Part II: conclusions,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,Mouse hepatitis virus ns2 prot,狂犬病病毒如何逃逸中枢神经系统先天性免疫反应？狂犬病病毒如何限制血脑屏障的打开？如何打开血脑屏障清除狂犬病病毒？,研究方向,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,狂犬病病毒如何逃逸中枢神经系统先天性免疫反应？研究方向嗜神经,41,谢 谢！,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,41谢 谢！嗜神经病毒躲避宿主先天性免疫反应的机制及应用,此课件下载可自行编辑修改，供参考！感谢您的支持，我们努力做得更好！,嗜神经病毒躲避宿主先天性免疫反应的机制及应用赵凌老师,此课件下载可自行编辑修改，供参考！嗜神经病毒躲避宿主先天性免,
