免疫组库测序技术介绍ppt课件

Immuno-Seq:exploringthediversityofimmunesystemJiankuiHeSouthUniversityofScienceandTechnologyofChina(SUSTC),Shenzhen,China.ThediversityofInvaders...BacteriaVirusesparasitessuchasfungi,protista,&wormsPhysicalinjury.TheImmuneSystemhttp://www.nobelprize.org.Innateimmunesystemageneralresponsetoanythingotherthanrecognized“selfcells”macrophages-abletoingestsmallforeigninvadersMastcells/basophilsreleasehistaminethatdilatesbloodvesselscausesredness.Adaptiveimmunesystemaspecificcounter-assaultagainsta“knownforeign”invader[previouslyrecognized].Key-lockEverymatureTcell(Bcell)expressesasingleTCR(BCR)withspecificityforalimitednumberofantigens.Tcellreceptor.HowmanyTcellsinyourbody?1.HumanbloodhasaboutonemillionTcellsineverymilliliter(ml).2.Theaverageadulthumanhasabout10litersofblood.3.Thereare10,000mlin10L.4,So,thereareaboutonemillionx10,000Tcellsinthebloodstream.Thiscomesto1010.About1/50ofallTcellsarefoundintheblood.Mostoftherestareinthespleenandlymphnodes.Somearealsofoundinthebonemarrow,liver,andothertissues.5.So,IwouldestimatethenumberofTcellsinthehumanbodytobeabout5x1011..BlymphocytedevelopmentHowantibodydiversityisgeneratedJanewayC.,Immunobiology,2004StemcellChromosome14,heavychain.BlymphocytedevelopmentHowantibodydiversityisgeneratedJanewayC.,Immunobiology,2004StemcellChromosome14,heavychainPro-Bcell.BlymphocytedevelopmentHowantibodydiversityisgeneratedJanewayC.,Immunobiology,2004StemcellBcellChromosome14,heavychainPro-Bcell.AntibodyrepertoireestimatesJointdiversity3-5bprandominsertion/deletionTheoreticalcalculation:Immunesystemonlyuses83geneSegmenttocode1014BCRsequencesHumanHeavychainVgenesegments~50HeavychainDgenesegments27HeavychainJgenesegments6VDJcombination50*27*6=8,000Potentialantibodydiversity~1014.DiversityandhealthyIntrinsicfactorsthatinfluencetheexpressionoftheTCRrepertoireinhumans:humanMHC(HLA)systemandTCRgenepolymorphisms,Extrinsicfactors:environmentalfactors,suchasinfectionandsuperantigens.InnormalindividualstheTCRrepertoireisstableandpolyclonal.clonalpopulationsarethehallmarkofmalignancy,clonaloroligoclonalpopulationsofTandBcellsmayoccurinnon-malignantconditionsincluding:humanimmunodeficiencyvirus(HIV)andEpstein-Barrvirus(EBV)infectionswithspecificityforvirus,intheelderlyinautoimmunityincommonvariableimmunodeficiency(CVID),inseverecombinedimmunodeficiency(SC1D),.TraditionalmethodstomeasurethediversityofTCR/BCRflowcytometryFluorescentlylabelledmonoclonalantibodiesagainstTCRVfamilygenesegmentsResolution:genesegmentlevelLimitedtoafewTCRVfamilygenesPCR+SangersequencingResolution:genesequenceLowthroughput.HighthroughputsequencingBcellsandTcellsStephenQuakelabScience,2009PNAS,2011Sci.Transl.Med,2013FirelabSci.Transl.Med2009PamerlabNat.Medicine2013HanlabPNAS,2010WarrenlabSci.Transl.Med2010GeorgioulabNat.Biotechnology2010.Thefirstuseofhighthroughputsequencing1200~3700uniqueantibodiesinafishStephenQuake.ThediversityofhumanTCR420,000uniqueCDR3nucleotideinafewmilliliterblood3*106uniqueCDR3isestimatedforanadultWearrivedatsurprisinglyhighminimalestimatesof100millionuniqueTCRBsequencesinnaïveCD4andCD8T-cellrepertoiresofyoungadults.Naïverepertoirerichnessmodestlydeclinedtwo-tofivefoldinhealthyelderly..Supersensitivityinminimalresiduediseasemonitoring(Leukemia)HighthroughputsequencingTCRcandetect0.001%frequencyoftumorcellsFlowcytometercandetect0.1%frequency..给免疫系统“拍照”“Immuno-Seq”技术使用新一代测序可以一次测出500万个TCR受体或者BCR的RNA序列。通过生物信息学方法分TCR/BCR的多样性和丰度，可以得知人体对感染，疫苗，和治疗的免疫反应状况。VDJN.Jiang,etal.,SciTranslMed.2013Feb6;5(171):171接受流感疫苗后.Immuno-SeqImmuno-Seq技术可以观察和 分析 定性数据统计分析pdf销售业绩分析模板建筑结构震害分析销售进度分析表京东商城竞争战略分析 T细胞和B细胞，达到前所未有的深度和特异性。在疾病监控，抗体生产，疫苗研究，健康体检等领域有广泛的应用。现在临床上有许多种检测可以监控你的健康状况，但是却没有一种被广泛接受的方法可以检测你免疫系统的各个方面。我们怎么可以得知一个人的免疫系统是很强健还是很虚弱？在人们受哮喘，过敏，关节炎折磨时，如何量化免疫系统的错误？斯坦福大学的一个研究团队在开发这样的检测方面迈出了第一步，他们的方法是：给免疫系统“拍照”。Therearenumerousteststogaugethehealthofyourheart.Butnosuchwidelyacceptedtestexistsforthemanypartsoftheimmunesystem.Howcanwetelliftheimmunesystemisstrongorweak?Orquantifyhowbadlyit’smalfunctioningwhenwesufferfromasthma,allergies,orarthritis?------FrancisCollins,NIHDirector2013年2月13日.ImmuneresponsetofluvaccineDay0Day7Day28FluVaccine:TIV&LAIVCollect5mlblood.Experimentflow.BroadVDJgenesegmentusagepatternVDJ.AsnapshotofimmunesystemDay7,aftervaccination.ElderlysubjectWeamplifiedantibodysequencesfromsinglesortedplasmaBcell.Wethenexpressedmonoclonalantibodiesaccordingtothesesequencesandverifiedtheirbindingtoeachofthethreevirusstrainsinthevaccine.ImmunereperotireisadynamicssystemDay0Day7Day28Vaccinespecificantibodylineage13%oftotalsequencesVaccinespecificantibodylineage0.01%oftotalsequencesElderlysubject.ReducedantibodydiversityinelderlyAge8-17Age18-32Age70-100MutationEachdatapointrepresentalineageComparisonof3subjectsSequencesinalineage.Reducedantibodydiversityinelderly70–100yearoldvolunteershadasignificantlylessdiversecollectionofantibodiesthantheyoungervolunteers.Thatis,aspeopleaged,theirimmunesystemhadamorelimitedrepertoireofantibodiestooffer.LabMembersPostdoc:ZhoufangLiTechnician:LuwenNingMengZhangStudents:JiajieGuoYinTongYingXuPengLiYaoChenRonghaiChengCollaborators:PinghuaLiu(BostonU),HengyeMan(BostonU)ZhigangLiu(ShenzhenU),ZhanhuiWang(SMU),ZhimingCai(2ndHospitalofShenzhen),XiaoLiu(BGI)XiaoPingChen(CAS),HuaZhang(SMU)Labweb:www.sustc-genome.org.cn.免疫组库测序服务提供商深圳市瀚海基因生物科技有限公司专注于提供人，小鼠，猴子等的TCR/BCR免疫组库测序网址：www.pacgeno.com.