冠狀病毒Coronavirusl論文-2006 Immunization of Mice with a DNA Vaccine Based on Severe Acute Respiratory Syndrome Coronavirus Spike Protein Fragme.pdf

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1、VIRALIMMUNOLOGYVolume19,Number3,2006?MaryAnnLiebert,Inc.Pp.518–524ImmunizationofMicewithaDNAVaccineBasedonSevereAcuteRespiratorySyndromeCoronavirusSpikeProteinFragment1BOZHAO,1NING-YIJIN,2RUI-LINWANG,2LI-SHUZHANG,2andYING-JIUZHANG1ABSTRACTAccordingtodatainGenBank,

2、ageneencodingSARSspikeproteinfragment1(S1)wassynthe-sized.Afterrecombinationwithanimmunostimulatorysequence(ISS),thegenewasclonedintotheplasmidpIREStoproducepIRES-ISS-S1.OnconfirmationoftheexpressionofS1proteinbyin-directimmunofluorescenceassay(IFA),afterthetransf

3、ectionofpIRES-ISS-S1intoBHK-21cells,theDNAvaccinewasrepeatedlyadministratedtoBALB/cmice.CD4andCD8spleenTlym-phocyteswereanalyzedbyflowcytometry(FCM)toevaluateTcell-mediatedimmuneresponses,theantigen-specificresponsesofTcellswereevaluatedbycytotoxicTlymphocyte(CT

4、L)assay,andthelevelofIgGinantiserafromimmunizedmicewasdeterminedbyenzyme-linkedimmunosor-bentassay.ResultsshowedthatthecountsofspleenCD4andCD8Tlymphocyteswereincreased,thattheTcell-mediatedimmuneresponsesshowedantigenspecificity,andthatIgGwassignifi-cantlyinduce

5、dwithDNAvaccinespIRES-ISS-S1andpIRES-S1attitersof1:320and1:160,re-spectively.Theseresultsarepromisingfortheprotectiveimmunizationofhumans.INTRODUCTIONtheexteriorsurfaceofcoronaviruses.TheSproteinofSARS-CoViscomposedofaknobunit(S1)andastemSEVEREACUTERESPIRATORYSYND

6、ROME(SARS)wasunit(S2),referringtotheN-terminalandC-terminalgly-firstreportedinlate2002inChina’sGuangdongcopolypeptidefragments,respectively,fromthecleavageProvince.Sincethen,ithasbecomeaworldwidethreatofprecursorSproteinbyspecialproteases(5,7,12).Thetopeople’sheal

7、th,andthediseasehasspreadtomoreS1unitofthespikeproteinislocatedontheoutersur-than30countriesandregions.TheSARSvirus,anovelfaceofthevirionandisbelievedtofunctionbybindingCoronavirus(SARS-CoV),iscomposedofasingle-totheexteriorreceptoronhostcells.Ithasbeenidenti-stra

8、nded,positive-senseRNAofapproximately29kb.Itsfiedasahostprotectiveantigenandusedasacandidategenomecontainsfivemajoropenreadingframesencod-vaccineagainst