A Novel High-Throughput Vaccinia Virus Neutralization Assay and Preexisting Immunity in Populations from Different Geographic Regions in China 英文参考文献.docVIP

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A Novel High-Throughput Vaccinia Virus Neutralization Assay and Preexisting Immunity in Populations from Different Geographic Regions in China 英文参考文献.doc

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A Novel High-Throughput Vaccinia Virus Neutralization Assay and Preexisting Immunity in Populations from Different Geographic Regions in China 英文参考文献

ANovelHigh-ThroughputVacciniaVirusNeutralization AssayandPreexistingImmunityinPopulationsfrom DifferentGeographicRegionsinChina QiangLiu1,2,WeijinHuang1,JianhuiNie1,RongZhu1,DongyingGao3,AijingSong1,ShufangMeng1, XuemeiXu2,YouchunWang1* 1Department ofCellBiology,NationalInstitutes forFoodandDrugControl,KeyLaboratory oftheMinistryofHealthfor ResearchonQualityandStandardizationof BiotechProducts,Beijing,China,2DepartmentofBiophysicsandStructuralBiology,InstituteofBasicMedicalSciencesChineseAcademyofMedicalSciences,Schoolof BasicMedicinePekingUnionMedicalCollege,Beijing,China,3BeijingBloodCenter,Beijing,China Abstract Background: Pre-existing immunity to Vaccinia Tian Tan virus (VTT) resulting froma largevaccination campaign against smallpoxpriortotheearly1980sinChina,hasbeenamajorissueforapplicationofVTT-vectorbasedvaccines.Itisessential to establish a sensitive and high-throughput neutralization assay to understand the epidemiology of Vaccinia-specific immunityincurrentpopulationsinChina. Methodology/Principal Findings: A new anti-Vaccinia virus (VACV) neutralization assay that used the attenuated replication-competentVTTcarryingthefireflyluciferasegeneofPhotinuspyralis(rTV-Fluc)wasestablishedandstandardized forcriticalparametersthatincludedthechoiceofcellline,viralinfectiondose,andtheinfectiontime.Thecurrentstudy evaluatedthemaintenanceofvirus-specificimmunityaftersmallpoxvaccinationbyconductinganon-randomized,cross- sectionalanalysisofantiviralantibody-mediatedimmuneresponsesinvolunteersexamined30–55yearsaftervaccination. TherTV-Flucneutralizationassaywasabletodetectneutralizingantibodies(NAbs)againstVacciniaviruswithouttheability todifferentiatestrainsofVacciniavirus.Weshowedthattheneutralizingtitersmeasuredbyourassayweresimilartothose obtainedbythetraditionalplaquereductionneutralizationtest(PRNT).Usingthisassay,wefoundalowprevalenceofNAb toVTT(7.6%)inindividualsbornbefore1980fromBeijingandAnhuiprovincesinChina,andwhenpresent,anti-VTTNAb titers were low