1、EnterovirusesChapter 24 Gastrointestinal Viruses p319-328 Family Group DiseasesEnteroviruses +ssRNARotavirus dsRNA diarrheaCalicivirus +ssRNA diarrheaAstrovirus +ssRNA diarrheaEnteric adenovirus(40,41) dsDNA diarrheaMedical Microbiology(2nd version) 臨床診斷病例 急性起病，發(fā)熱，手掌或腳掌部出現(xiàn)斑丘疹和皰疹，臀部或膝蓋也可出現(xiàn)皮疹。皮疹周圍有炎性紅

2、暈，皰內(nèi)液體較少；口腔粘膜出現(xiàn)散在的皰疹，疼痛明顯。部分患兒可伴有咳嗽、流涕、食欲不振、惡心、嘔吐和頭疼等癥狀。重癥病例：1.有XXX病的臨床表現(xiàn)的患者，同時(shí)伴有肌陣攣，或腦炎、急性遲緩性麻痹、心 肺衰竭、肺水腫等。2. 流行地區(qū)的嬰幼兒雖無XXX病典型表現(xiàn)，但有發(fā)熱伴肌陣攣，或腦炎、急性 遲緩性麻痹、心肺衰竭、肺水腫等 neurology ,Vol 9, November 2010什么?。咳绾芜M(jìn)一步確診？預(yù)防和治療？Classification of Viruseshttp:/www.nlv.ch/Virologytutorials/Classification.htm The Baltim

3、ore classification clusters viruses into families depending on their type of genome/viralzone/all_by_species/254.htmlClassification and Genomes of Human VirusesGROUPFAMILYEXAMPLESGENOME SIZE ()I. (ds DNA)PapillomavirusesHPVs8 KbpAdenovirusesAdenoviruses40 KbpHerpesvirusesHSV-1 & 2, VZV, EBV, CMV130

4、- 230 KbpPoxvirusesSmallpox (Variola) and Vaccinia150 - 250 KbpII. (ss DNA)ParvovirusesParvovirus5 KbIII. (ds RNA)ReovirusesRotavirus22 Kbp (11 pieces)IV. (+ ss RNA)PicornavirusesPoliovirus, Rhinoviruses, Hep. A8 KbCoronavirusesCoronavirus28 KbTogavirusesRubella12 KbFlavivirusesYellow Fever, West Ni

5、le, Hep. C10 KbV. ( - ss RNA)RhabdovirusesRabies11 KbParamyxovirusesMeasles and Mumps14 KbOrthomyxovirusesInfluenza A and B12 Kb (8 pieces)BunyavirusesHantavirus18 Kb (3 pieces)ArenavirusesLassa12 Kb (2 pieces) FilovirusesEbola and Marburg19 KbVI. (RNA rev. trans.)RetrovirusesHIV and HTLV10 KbVII (D

6、NA rev. trans.)HepadnavirusesHep. B 3 Kbp Genera of PicornavirusesEnterovirus PolioCoxsackie A and BEchoOther enteroviruses Diseases of the human (and other) alimentary tract (e.g. polio virus)RhinovirusDisease of the nasopharyngeal region (e.g. common cold virus)CardiovirusMurine encephalomyocardit

7、is, Theilers murine encephalomyelitis virusAphthovirusFoot and mouth disease in cloven footed animalsHepatovirusHuman hepatitis virus AOthersDrosophila C virus, equine rhinoviruses, cricket paralysis virusCategories of EnterovirusesViral meningitis, rash,ARD4 types(68-71)Other EnterovirusesViral men

8、ingitis, with orchitis32 typesEchovirusesViral meningitis, but no orchitis6 types (B1-B6)Coxsackie B virusesViral meningitis plus, rash, ARD, myocarditis, orchitis23 types ( A1-A22, A24)Coxsackie A virusesAsymptomatic infection, viral meningitis, paralytic disease, poliomyelitis3 typesPoliovirusesCl

9、inical DiseasesSerotypesVirusProperties of enterovirusesPropertyEnterovirusesSize (nm)Capsid form PolypeptideRNA typeRNA molecular weightAcidOptimal temperature for growth(oC)Density in caesium chloride (g/m)22-30Icosahedral VP1, VP2, VP3, VP4SS-PS2000,000-2600,000Stable*371.34*VIRION and GENOMEThe

10、polyprotein products and their main functions.Whitton,2005,Nature Review MICROBIOLOGYTransmissionFecal oral route: poor hygiene, dirty diapers( especially in day-care settings) Ingestion via contaminated food and waterContact with infected hands Inhalation of infectious aerosolsViral infection and t

11、issue tropismsHuman diseases caused by enteroviruses PolioCox A Cox BEchoOtherAsymptomatic infectionYes Yes YesYesYes MeningitisYes Yes YesYesYes ParalysisYes Yes YesYesNo Febrile exanthemsNo Yes YesYesYes Acute respiratory diseaseNo Yes YesYesYes MyocarditisNo Yes YesYesNo OrchitisNo No YesYesNo Li

12、fe cycle of the picornavirusWhitton,2005,Nature Review MICROBIOLOGYIRES-mediated translation initiationNATURE Reviews| Molecular cell Biology，2010，Vol 10，113-127ImmunityAntibody is the major protective immune response to the enteroviruses . Secretory antibody can prevent the initial establishment of

13、 infection in the oropharynx and gastrointestinal tract, and serum antibody prevents viremic spread to the target tissue and therefore disease.Cell-mediated immunity is not usually involved in protection but may play a role in pathogenesis.Signaling pathways activated by RNA leading to interferon pr

14、oduction in virus-infected cellsCharles E. Samuel, JBC, 2007, may25, 15313-15314Prevention and therapyPrevention: VaccinesTherapy: anti-viral drugsLaboratory DiagnosisCultureSerologyGenome PoliovirusEgyptian hieroglyph indicates presence since 1400 BC1840 - Heinle characterizes poliomyelitis “grey m

15、arrow” in GreekPoliomyelitis:Former-president Franklin D. Roosevelt in United StatesVictims of paralytic polio Clinical SyndromesAsymptomatic illness: 90%Abortive poliomyelitis, the minor illness: 5% infected peopleNonparalytic poliomyelitis or aseptic meningitis: 1%-2% of patients with poliovirus i

16、nfections.Paralytic polio, the major illness: 0.1% to 2%of persons with poliovirusIron Lung:Paralyzed child in an iron lungPoliovirus:Family:PicornaviridaeGenera:Enterovirus types 1, 2 and 3Dissemination of PoliovirusAxonal transport of PoliovirusOhka, S and Nomoto, A. (2001) Trends in Microbiology

17、9(10) 501Poliovirus Genome Kitamura N , Wimmer E et al:Nature. 1981 Jun 18;291(5816):547-53. Poliovirus receptor identificationIntact poliovirus infects human cells but not murine cellsMurine cells transfected with human DNA are susceptible to infection with intact poliovirusHumanDNAMendelsohn C, Wi

18、mmer E, Racaniello VR et al:Proc Natl Acad Sci U S A. 1986 Oct;83(20):7845-9. Poliovirus Receptor/CD155/Necl5COOHVC1C2NH2NH2TransmembraneDomainPotential N-linkedglycosylation sitesMendelsohn CL, Wimmer E, Racaniello VR et al: Cell. 1989 Mar 10;56(5):855-65. Interaction of the poliovirus receptor wit

19、h poliovirusJ Virol. 2003 Apr;77(8):4827-35. Proc Natl Acad Sci U S A. 2000 Jan 4;97(1):79-84. Virology 1994,203:344-356V-domain structure of PVR/Necl5 and Necl1 Life cycle of poliovirus:ImmunitysIgA and neutralizing antibody (IgG, IgA, IgM) persist for life spanReciprocal virus antibody titer512128

20、32821Serum IgGSerum IgGSerum IgMSerum IgMNasal and duodenal IgANasal IgASerum IgASerum IgADuodenal IgADaysVaccinationVaccination48489696Killed (Salk) VaccineLive (Sabin) VaccineLab DiagnosisDefinitive diagnosis is made by isolation of the virus from stool, CFS, oropharyngeal secretionsCell culture i

21、nvolves fibroblastic MRC-5 cellsCPE is usually evident within 36 hoursSerotyping is based on neutralization of CPE by standardized antisera using intersecting pool followed by specific sera.ELISAIFAneutralizing TestCFTPreventionBoth oral polio vaccine( OPV live, attenuated， Sabin, 1957) and inactiva

22、ted poliovirus vaccine(IPV，Salk, 1954) are availableIPV is used for adult immunization and Immunocopromised patientsVaccines:1954 - Salk vaccine(IPV)1960 - Sabin vaccine(OPV)1959-1962(China)Institute of Medical Biology, CAMS & PUMCReported cases per 100000 population1001010.10.0010.01195019601970198

23、01990Inactivated (Salk) vaccineOral vaccineCases per 100,000 population United StatesEpidemiology before and after Vaccine-use(1)1000010001001010Reported cases195019551960196519701975Killed (Salk) vaccineTotal casesSweden and FinlandEpidemiology before and after Vaccine-use(1)Epidemiology before and

24、 after Vaccine-use(2)Global Polio Eradication1988 World Health Assembly calls for global polio eradication by 2000. Achieving Certification: three WHO regions which had been certified polio free by the end of 2003 the Region of the Americas (1994), the Western Pacific Region (2000) and the European

25、Region (2002). China:2008-2010Problems: poliovirus can circulate invisibly without causing disease or symptoms OPV vaccine might it self trigger a devastating outbreak and can circulate silently in the population and revert, “persistent shedders”-need new vaccine! the virus can be synthetically synt

26、hesized -From: Science, 26 March 2004 Ronit Sarid, 2004 Threat of bioterrorism remaining Laboratory strainsContributions of Poliovirus to life sciences research Isolation of Poliovirus John Enders and the Nobel Prize Discovery of viral & cellular IRESes-IRES model of protein translationPoliovirus 5-

27、NCR StructureIIIIIIIVVVIVIIVPg288137158188222236443471538585620743AUGRNAsynthesisIRESAttenuationAdapted from Macadam et al. (1994) Trends in Micro., 2, 449-454Sung K. Jang and Eckard Wimmer:Journal of Virology.1988,62(8):2636-2643.Jerry Pelletier and Nahum Sonenberg:Nature.1988,334:320-325.Three Cel

28、lular IRESesOncogene (2004) 23, 32003207 Protein-primed RNA synthesisPaul AV, van Boom JH, Filippov D, Wimmer E :Nature. 1998 May 21;393(6682):280-4. Intergeneric poliovirus recombinants for the treatment of malignant glioma Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6803-8.PV/HCV chimeric virus as

29、 model for anti-HCV drug screeningProc Natl Acad Sci U S A. 1996 Feb 20;93(4):1412-7.A model for RNA viruses dominant inhibitor screening -Validate or predict targets for antiviral therapy with reduced risk for drug resistance Nat Genet. 2005 Jul;37(7):701-9. Poliovirus and synthetic lifeCell-free,

30、de novo synthesis of poliovirus-Molla A, Paul AV, Wimmer E. Science. 1991 Dec 13;254(5038):1647-51. Dear Xiaozhong,The lab has changed of course, but you would recognize still many familiar faces. Aniko, Jeronimo, Steffen, Shan, JoeAnn, and Connie, for example. We are still working mainly on poliovi

31、rus although we have also developed a program on hepatitis C virus. The most recognized work of the last years, however, was the chemical synthesis of poliovirus in the absence of natural template, which we published in 2002. Perhaps you have heard about it. It caused trouble and praise. With warm r

32、egards to you and to your familyfrom Astridand from everybody in the labyoursEckardBIOTERRORISMEV71Pathological findings in enterovirus 71 encephalitisneurology ,Vol 9, November 2010Epidemic of HFMDUp to June 2011, there are 303594 reports and 97 deaths.reports deaths衛(wèi)生部數(shù)據(jù)Enterovirus 71 genotypic su

33、bgroups reported to be circulating in the Asia-Pacific region between 1973 and 2008, by yearStructure biology of EV71Structure of EV71 3CproCui,2011,JMBStructure of EV71 RdRp 3DproWu,2010,Protein CellCrystal structure of Human Enterovirus 71Wang x x, et al,. Nat structure& molecular biology, 2012A s

34、ensor-adaptor mechanism for enterovirus uncoating from structures of EV71Pavel, et al,. Science, 2012Crystal structure of Human Enterovirus 71Receptors identified for hand, foot and mouth virusTyrosine Sulfation of the Amino Terminus of PSGL-1 Is Critical for EV71 InfectionNishimura,2010, Plos Patho

35、gensIdentification of the Human SCARB2 Region That Is Important for Enterovirus 71 Binding and Infectionamino acids 142-204Yamayoshi,2011,JVITransgenic mouse model for the study of enterovirus71 neuropathogenesisExpression profiles of hSCARB2 in human and hSCARB2-Tg10 mouse tissuesKen Fujii, et al,.

36、 PNAS, 2013The Signal Pathway of EV71 infection of SK-N-SH.Proteins interact with the enterovirus 71 5untranslated region and participates in virus replicationLin,2008, Journal of General VirologyJournal of Medical Virology 2006,78:780-786.Journal of Immunology 2011,5(187):2201-2212.Virology journal

37、,2011,8(1):61.Scand J Infect Dis, 2010,42(11-12):931-935EV71T cells apoptosisInhibits TNF-Mediated Activation of NF-B (EV71 2C protein)degradation of TRIM38, a potential E3 ubiquitin ligaseSignificant elevation of B cells Efficacy, safety, and immunology of an inactivated alum-adjuvant enterovirus 7

38、1 vaccine in children in China: a multicentre, randomised, double-blind, placebo-controlled, phase 3 trialZhu F C et al,. The lancet, 2013Vaccine design of Enterovirus71Production of EV71 vaccine candidatesChong pele, et al,. Human vaccine & Immunotherapeutics, 2012Coxsackie VirusEnteroviral pathogenesis of type 1 diabetes Discov Med. 2010 Aug;10(51):151-60. Engineering microRNA responsiveness to decreasevirus pathogenicityTissue-specific destabilization of viral genomes by miRNA target insertion pro