1免疫学概论及分子生物学在免疫学中的应用

FangLi,GuizhiCheng,HongQu,BingWang,JieMa,ShiyingwangDept.ofImmunologyCollegeofbasicmedicalsciencesDalianMedicalUniversityTel:86110294:ImmunologyisthestudyofimmunityinthisbroadersenseandofthecellularandmoleculareventsthatoccurafteranorganismencountersmicrobesandotherforeignmacromoleculesTheImmuneSystemisthenameofacollectionofMolecules,CellsandOrganswhosecomplexinteractionsformanefficientsystemthatisusuallyabletoprotectanindividualfrombothoutsideinvadersanditsownalteredinternalcells.中枢免疫器官（Centrallymphoidorgan）淋巴细胞发育成熟的场所骨髓（bonemarrow）、胸腺（thymus）免疫器官immuneorgansT细胞免疫缺陷先天性胸腺发育不全（DiGeorge综合症）Thisisametaphase（中期）cellthathasbeenhybridizedwiththeprobeforDiGeorgewhichiscausedbyamicrodeletiononchromosome22.Theprobeisadual-colormixtureoftwoseperateprobesforchromosome22.Thegreensignalisaninternalcontrolandislocatedat22q13.Itallowsforquickidentificationofboth#22chromosomes.TheredsignalislocatedattheDiGeorgeregionat22q11.2.Sinceboth22'shavetheredsignalinthiscellthereisnotamicrodeletionwithintheDiGeorgeregionandthispatientwouldnothaveDiGeorgesyndrome.无胸腺的裸鼠是T细胞选择性缺陷的天然动物模型Athymic,ornude,mouse(nu/nu:onchromosome11)Thisdefectleadstoabsenceofthethymusoravestigialthymusandcell-mediatedimmunodificiency.Cannoteasilysurvive,mortality:100%within25weeks;50%within2weeksafterbirth.SkeletalpreparationsfromwildtypeandTbx1mutantembryos.Themutantembryoontherighthasabnormalitiesinthepharyngeal(咽的)arch-derivedskeletalstructuresinthehead.ThismutantprovidesamousemodelofthehumanDiGeorgesyndromeAthymicnudemicearekeptbehindabarrierwithHEPAfilteredair;autoclavedfee,bedding,waterandcages.AresearchassistantusesasepticprocedurestoprotectthesemicefromorganismsintheenvironmentTumorsformedbyRat1cells肿瘤组织的原位移植（SOI）取皮下种植生长的前列腺癌肿瘤1mm×1mm的组织碎块麻醉小鼠显微外科手术裸鼠下腹部做一1cm的纵切口，暴露前列腺将1块PC-3-GFP组织碎块植入裸鼠前列腺上20天25天30天CONTROL-3EXPERIMENT-3finalp.s：桔黄色空心箭头所指为转移的腰淋巴结；桔黄色实心箭头所指为原发肿瘤；白色空心箭头所指为肠系膜淋巴结转移；白色实心心箭头所指为胰腺转移CONTROL-3EXPERIMENT-3final外周免疫器官（Peripherallymphoidorgan）淋巴细胞定居和发挥功能的场所脾（spleen）、淋巴结（lymphoidnodes）黏膜相关淋巴组织（MALT）单核巨噬细胞（monocytesandmacrophages）免疫细胞immunecells抗体（antibody）主要组织相容性复合体（MHC）T细胞受体TCR免疫分子immunemolecules12345免疫记忆——疫苗与抗感染免疫免疫记忆（Immunologicalmemory）经历了某种微生物感染之后短时间内一般不会再次被同一种微生物感染。这种现象称为免疫记忆。Immunologicalmemorymakesvaccinationagainstinfectiousdiseasespossible!疫苗（VACCINE）Mycobacteriumtuberculosis.脊髓灰质炎Polio狂犬病Rabies天花Smallpoxvirus白喉diphtheriaHepatitisAHepatitisB接种疫苗后21天内的反映情况免疫记忆ImmunologicmemoryAB白喉vaccineDEAnti-白喉antibody白喉BmcellAnti-白喉antibodyagainst白喉白喉Bmcell获得性免疫应答免疫缺陷肿瘤变态反应(超敏反应)自身免疫性病移植免疫临床免疫及免疫病理——Hypersensitivity超敏反应Immunodeficiency免疫缺陷Autoimmunediseases自身免疫病Canceroftheimmunesystem免疫系统肿瘤TumorImmunology肿瘤免疫IHypersensitivity超敏反应Eczema湿疹AnatopiceczemareactiononthebackofakneeofachildallergictoriceandeggsAsthmaDC,Th2,B,MastcellEpithelium,EosIL-4,IgEAllergenSerumsicknessPatientswithhypersensitivityvasculitiswithtypicalskinrash.T、B细胞联合免疫缺陷SevereCombinedImmunodeficiency，SCIDIIImmunodeficiency免疫缺陷HIV感染淋巴细胞Immunodeficiency免疫缺陷HIVbuddingfromhumanlymphtissue(TEMx133,335)Immunodeficiency免疫缺陷LesionsonthestomachofapatientwithKaposi'ssarcomaFigure-Kaposi'sSarcomaKaposi'ssarcoma(skin).SkinshowingAIDS-associatedKaposi'ssarcomaImmunodeficiency免疫缺陷Hairyleukoplakia（黏膜白斑）oftongueinAIDSOralthrush鹅口疮.CandidaandherpessimplexinAIDSSevereangularcheilitisinAIDS口角炎Immunodeficiency免疫缺陷RA患者手指关节实质性损伤IIIAutoimmunediseases自身免疫病SLE患者脸部的蝴蝶斑Autoimmunediseases自身免疫病Systemiclupuserythematosis系统性红斑狼疮(Ach:acetylcholine)乙酰胆碱ankylosingspondylitis(B27),强直性脊柱炎Destructionofpancreaticislet-cellsbyinfiltratingT-cellsintheNonobesediabetic(NOD)mouseApplicationofMolecularBiologyinImmunologyMOLECULARBIOLOYSeektoexplaintherelationshipsbetweenthestructureandfunctionofbiologicalmoleculesandhowtheserelationshipscontributetotheoperationandcontrolofbiochemicalprocesses.Practicalapplicationsinthedevelopmentofnewandsafeproductssuchastherapeutics,vaccinesandfoodstuffs,andinthediagnosisofgeneticdiseaseandingenetherapyModernmolecularbiology:一、新细胞因子的cDNA克隆技术ProteinFMVL……..cDNAlibraries........AUGCCUAAUCCU……mRNATACGGATTAGGA……ProbeDesigningSpeculating.......TransfertonitrocellulosePlating32PDigColonyhybridizationPositivecolony传统的蛋白质到核酸途径差异显示技术克隆细胞因子利用同一细胞在不同条件下表达免疫分子的差异克隆新细胞因子，例如T细胞活化前低表达细胞因子，活化后则高表达细胞因子，利用DD-PCR、RDA、SSH等技术可克隆T细胞活化后高表达的基因，其中可能含有新细胞因子cDNA.(-)(+)现代的核酸到蛋白质途径Interleukin-10.Chain:l.(150residues)-[9helices]Cytokinesynthesisinhibitoryfactor.Interleukin-10receptoralphachain.Chain:rFragment.(205residues)-[2helices,12strands]:extracellulardomain,(-)IL-10(+)PHAExpressingmRNAcDNAReveretranscript第二轮杂交，加新的变性driver,退火加引物+PCR扩增回收片段，克隆化PHA刺激的U937细胞IL-10抑制的U937细胞PHA刺激的U937细胞TestercDNAwithadaptor-1DrivercDNA(超量)TestercDNAwithadaptor-2第一轮杂交末端补齐SuppressionSubtractiveHybridizationSSH示意图GTTCCCAATCTGAAGTGAAGCCGAGCTGGGCGAGAAGTAGGGGAGGGCGGTGC