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1、METHODSINMOLECULARMEDICINETMHepatitisBandDHepatitisBandDProtocolsProtocolsVVolumeII:Immunology,olumeII:Immunology,ModelSystems,ModelSystems,andClinicalStudiesandClinicalStudiesEditedbyEditedbyRobertK.Hamatake,RobertK.Hamatake,PPhhDDJohnsonY.N.Lau,JohnsonY.N.Lau,MDMD1StudyingHostImmuneRe
2、sponsesAgainstDuckHepatitisBVirusInfectionDarrenS.Miller,EdwardM.Bertram,CatherineA.Scougall,IevaKotlarski,andAllisonR.Jilbert1.IntroductionTheduckhepatitisBvirus(DHBV)isaspecies-specificvirusthatcauseseithertran-sient(acute)orpersistentinfections,primarilyinhepatocytesintheliver,withrel
3、easeofhightitersofinfectiousvirionsandnoninfectious“empty”surfaceantigenparticlesintothebloodstream.Becausehepadnavirusreplicationisnoncytolytic,cell-mediatedimmune(CMI)responsestoviralantigensarethoughttoberesponsiblefortheclearanceofvirusfrominfectedcellsandfortheliverdamageseenintran
4、sientandpersistentinfections.Thisispresumedtooccurviaadirect,cytolyticeffectofviralantigen-specificcytotoxicTlymphocytes(CTLs)oninfectedhepatocytes,orviathenoncytopathicactionofinflam-matorycytokines.Inaddition,neutralizingantibodieshavebeenshowntopreventinfectionbyblockingtheabilityofvir
5、usparticlestobindtoreceptorsontargetcells.DHBV-infectedducksandwoodchuckhepatitisvirus(WHV)-infectedwoodchucksarethemostwidelyacceptedandfrequentlyusedanimalmodelsforthestudyofviralreplication,infectionoutcomes,andthepathogenicmechanismsrelatedtohumanhep-atitisBvirus(HBV)infection.Useof
6、theDHBVmodelhasallowedustostudytheeffectsofviraldose,age,andinoculationrouteonthecourseofDHBVinfection(1–4)andtheeffectofimmunizationwithvariousformsofvaccineonalltheseparameters(5).However,untilrecently,studiesoftheimmuneresponsetoDHBVinfectionhavebeenhamperedbytherelativelypoorcharact
7、erizationoftheducklymphoidsystemandthelackofappropriatereagents.ThischapterdescribesanumberofassaysthatallowstudyofcomponentsoftheduckimmunesystemandthecellularandhumoralimmuneresponsestoDHBVinfection.Thechapterhasbeendividedintothreesectionsthatinclude:1.Purificationandcharacte
