Adenoviridae课件.ppt

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1、AdenoviridaeMolecular Virology HISTORYqfirst isolated in 1953(Rowe,et al.(1953),Proc.Soc.Exp.Biol.Med.84:570573)from tonsils and adenoids of childrenqsimilar viral agents were isolated from febrile military personnel with respiratory illnessesqadenoid degeneration(AD),adenoid-pharyngeal conjunctival

2、(APC),and acute respiratory disease(ARD)agentsq1956 1956 ADENOVIRUS ADENOVIRUSnormal cell culturepassagecytopathic effectvirionsTAXONOMYGROUPGroup I dsDNAFAMILYAdenoviridae(Greek,adenos,“gland”)GENUSMastadenovirus(Greek,mastos,“breast”)Aviadenovirus(Latin,avis,“bird”)Atadenovirus(English,adenine and

3、 thymine)Siadenovirus(English,sialidase)Ichtadenovirus(Greek,ichthys,“fish”)TYPE SPECIESTYPE SPECIESOvine Adv DHuman Adv CFowl Adv AFrog AdvSturgeon Adv AGENOMEqlinear dsDNA;30-36 Kbqinverted terminal repeat(ITR)sequences at 3-end;30-200 bpqlinear DNA forms a loop due toterminal protein(covalently l

4、inked to each 5 end)and terminal base pairingqall genes are transcribed by host RNApol II except VA gene which istranscribedbyRNApolIIIqmRNAs are polycistronic and are differentiated by alternative splicing and use of different poly(A)sitesSTRUCTURE qnon-enveloped,icosahedral capsid;90-120 nmq20 equ

5、ilateral triangular faces,12 verticesqeach face has 12 identicalhexonshexonsand each vertex has 1pentonpenton;qeach penton is attached to afiber proteinq12 x 20=240 hexons;also 12 pentonsqtotal of 252 capsomersCAPSID Protein II Protein III(penton base)-penetrationCAPSID Protein II Protein III Protei

6、n IIIa -penetration CAPSID Protein II Protein III Protein IIIa Protein IV Protein VI -hexon minor polypeptide -stability/particle assemblyCAPSID Protein II Protein III Protein IIIa Protein IV Protein VI Protein VIII Protein IX -hexon minor polypeptide -stability/particle assemblyCAPSID Protein II Pr

7、otein III Protein IIIa Protein IV Protein VI Protein VIII Protein IX -hexon minor polypeptide -stability/particle assemblyCORE Protein V -links DNA to penton base -histone-like,packaging CAPSID Protein II Protein III Protein IIIa Protein IV Protein VI Protein VIII Protein IX -hexon minor polypeptide

8、 -stability/particle assemblyCORE Protein V -links DNA to penton base -histone-like,packaging Protein VII Protein X()-packaging CAPSID Protein II Protein III Protein IIIa Protein IV Protein VI Protein VIII Protein IX -hexon minor polypeptide -stability/particle assemblyCORE Protein V -links DNA to p

9、enton base -histone-like,packaging Protein VII Protein X()55kD Terminal protein -genomic replicationCAPSID Protein II Protein III Protein IIIa Protein IV Protein VI Protein VIII Protein IX -hexon minor polypeptide -stability/particle assemblyCORE Protein V -links DNA to penton base -histone-like,pac

10、kaging Protein VII Protein X 55kD Terminal protein dsDNAATTACHMENT&ENTRYCD46 for Human Adv subgroup BEARLY TRANSCRIPTION&TRANSLATION12 hrASSEMBLY&RELEASE100K protein facilitates folding and assembly of hexon trimersProtein VI stabilizes capsid and facilitates hexon importationIVa2,L1 52/55K,L4 22kD

11、promote viral DNA packagingCleavage of precursors of VI,VII,VIII,X by viral proteaseViral escape and spread of progeny virus by:1)L3 protease cleavage of cellular cytokeratin2)ADP kills cells3)free fiber trimers released from infected cells interfere CAR oligomerization at tight junctions Immune Eva

12、sion1.Inhibition of IFN functions-VA-RNA and E1A binds to PKR2.Inhibition of TNF and Fas-mediated apoptosis-E1B19K,E314.7,E310.4K/14.5K3.Downregulation of surface class I MHC-E3gp19,E1A retention of MHC I in ERE3 proteins and their functionsKnipe and Howley.Fields Virology.5th ed.orf6/7orf6orf4orf3o

13、rf2orf1E4Modulates E2FInteracts with E1B 55Kfacilitating RNAmetabolismBinds to DNA PKInhibits E1Aactivation of E2FBinds to DNA PKInteracts with E1B 55KRelocates nuclear pods?Facilitates transformationE4 proteins and their functionsJournal of General Virology(2000),81,25732604Journal of General Virol

14、ogy(2000),81,25732604Oncogenic potential of human adenovirusesKnipe and Howley.2007.Fields Virology.5th ed.CLINICAL FEATURES OF DISEASEENTRYENTRYSPREADSPREADRESPIRATORY INFECTIONSRESPIRATORY INFECTIONSpharyngitispertussus pneumoniaacute respiratory diseaseOCULAR INFECTIONSOCULAR INFECTIONSconjunctiv

15、itiskeratoconjunctivitisGASTROINTESTINAL INFECTIONSGASTROINTESTINAL INFECTIONSgastroenteritishepatitismeningoencephalitismyocarditishemorrhagic cystitisAdenovirus diseases,associated serotypes,hosts and clinical specimens for diagnosisAdenovirus diseases,associated serotypes,hosts and clinical speci

16、mens for diagnosisZuckerman,et al.2009.Principles and Practice of Clinical Virology.6th ed.Diseases of Domestic Animals Associated with AdenovirusesDiseases of Domestic Animals Associated with AdenovirusesMurphy,et al.Veterinary Virology.3rd ed.DIAGNOSIS1.DIRECT METHODSa.Viral isolation human epithe

17、lial cell linesb.Histopathology enlarged nuclei with basophilic inclusionsc.Direct antigen detectioni.IFAii.EIAiii.Immunochromatographyiv.IHCd.Direct particle detectioni.EM acute gastroenteritise.Direct genome detectioni.PCR,real-time PCR2.INDIRECT METHODSa.Serology IgM/IgGI.GENE THERAPY VECTORS-gen

18、e to correct genetic defectII.CANCER THERAPY VECTORS-gene induces cell death III.VACCINE VECTORS-gene is antigenUSE OF ADENOVIRUSES AS:Types of Adenovirus Vectors1)Replication-defective vectors-one or more viral genes deleted2)Replication-competent vectorsReplication-defective vectorsFIRST GENERATIO

19、N VECTORS-E1 deletedSECOND GENERATION VECTOS-E1,E2,E4 deleted-helper-dependent vectorKnipe and Howley.Fields Virology.5th ed.Replication-competent vectorsKnipe and Howley.Fields Virology.5th ed.ADENOVIRUSES AS VECTORS FOR VACCINATION AND GENE THERAPHYAdvantagesqcan be grown to produce stable,high ti

20、ter stocks;qcan infect broad range of tissues including nondividing cells;qrarely integrate into the host chromosome Disadvantagesqduration of the expression of the transgene will be limitedqchances of the vector to bind to non-target cellsqsize of foreign gene is limitedJournal of General Virology(2000),81,25732604Thanks for listening