博士生课程1(1)-免疫学概述.ppt

单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,The Origin of Immune Concept,The term“Immunity”,=Latin word“,Immunitas,”=Protection from legal prosecution(Roman senators),Biological definition=Protection from infectious diseases,2.The concept of immunity=existed in ancient Greek&Chinese=the experienced view,“天花”又名痘疮,中国从宋朝起用人痘接种预防天花,The medical view of immunity=Edward Jenner(1796),Observation,=Milkmaids generally get,No Smallpox,Hypothesis,=Pus from,vaccinia,(cowpox),=Protect milkmaids from smallpox,Test,=Inoculate materials from cowpox pus,=Protect a young boy from smallpox,(Protective immunity),The vaccination against smallpox,Exudate from a cowpox pustule on the hand of milkmaid Sarah Nelmes was inserted into scratches on the arms of James Phipps,May 14,1796,.,Edward Jenner,Eradication of smallpox,200 years,after Jenner,WHO announce,smallpox eradicated,1965,1970,1975,1980,Countries with,more than one,smallpox case,per month,30,15,0,免疫的基本功能,功能,正常表现（有利）,异常表现（有害）,免疫防御,immune defence,抗病原微生物,的侵袭,超敏反应,易受感染或免疫缺陷病,免疫稳定,immune homeostasis,清除损伤、衰老、死亡细胞,自身免疫性疾病,免疫监视,immune surveillance,清除突变或畸变的恶性细胞,恶性肿瘤,免疫系统,(immune system),：,机体执行免疫功能的组织、器官、细胞和分子构成免疫系统。,The four kinds of pathogens that cause human disease,常见的病原微生物,Overview of immune responses,固有免疫,（,innate immunity,）,是机体抵御病原微生物入侵的第一道防线，并启动和参与适应性免疫应答。,天然免疫（,natural immunity),或非特异性免疫,(nonspecific immunity),，是个体出生时就具有的免疫力，通过遗传获得，是生物在长期进化过程中逐渐形成的，其针对外来异物的范围广，反应迅速，其应答模式和强度不因与病原微生物的反复接触而改变。,固有免疫应答的作用时相：,瞬时固有免疫应答阶段、早期固有免疫应答阶段、适应性免疫应答诱导阶段,固有免疫系统的组成,屏障,细胞,分子,皮肤黏膜屏障：,物理、化学、微生物,血,-,脑屏障、血,-,胸腺屏障,血,-,胎屏障、气,-,血屏障,巨噬细胞、中性粒细胞、树突状细胞、,T,细胞、,NK,细胞、,NKT,细胞、,B1,细胞、肥大细胞、嗜碱性粒细胞和嗜酸性粒细胞等。,抗菌肽、溶菌酶、急性期蛋白、补体、细胞因子和黏附分子、,Epithelial barriers prevent the entry of microbes,固有免疫细胞,Phagocyte,NK,ILLs,（固有样淋巴细胞）,DC,MC,BasophilEosinophil,T,细胞,NKT,细胞,B1,细胞,Monocyte-macrophage,Neutrophil,肺部巨噬细胞吞噬大肠杆菌,Phagocytosis by innate immunity,Leukocyte recruitment to sites of inflammation,or DC,固有性免疫分子,指体表分泌液以及血浆和其它体液中能够识别或攻击病原体的可溶性分子。,抗菌肽,antimicrobial peptides,溶菌酶,lysozyme,急性期蛋白,(acute phase proteins,APP),脂多糖结合蛋白（,LBP,）,血清淀粉样蛋白（,SAP,）,甘露糖结合蛋白（,MBP,）,C,反应蛋白等（,CRP,）,补体,细胞因子和黏附分子,Complement activation pathways,Elie Mechnikoff:,The Pioneer of Innate Immunity,1.The Discovery of Phagocytes&Phagocytosis,2.The Nobel Laureate in Medicine 1908,Adopted from Nature Immunology,July 2008,The development of modern Immunology in 20,th,century,mainly centers on understanding the Adaptive,Immune System.,Charles A.Janeway,M.D.,Yale Univ.,The“Renaissance”of innate immunity,In 1989,Janeway=Immune recognition of microbes=Detection of conserved molecular patterns,referred to,PAMPs(Pathogen-Associated Molecular Patterns)with features:,1.Invariant among a given class of,microbes.,2.Have essential roles in microbial,physiology.,3.Recognized by receptors of the,innate immune system,called,PRRs(Pattern-Recognition,Receptors).,4.Innate immunity regulates adaptive,immunity,Julie A.Hoffmann,Ph.D.,Strasbourg,France,The“Renaissance”of innate immunity,In 1996,Hoffmanns group,Toll functions as a PRR,in Drosophila,Key concepts in innate immunity,1.The innate immune system mainly recognizes common,structures shared by classes of microbes,=,Pathogen,Associated Molecular Patterns(PAMPs),e.g.,LPS,Peptidoglycan,Microbial DNA&RNA.,2.Host receptors that recognize PAMPs are called,Pattern-,Recognition Receptors(PRRs),which are encoded in,“Germline”DNA=limited Diversity.,3.Innate immunity not only provide the first line of,defenses but link to the program of adaptive immunity.,4.PRRs may also recognize components from injured or dead,host cells=,Autoimmune diseases,Pattern,Lipopolysaccharide(LPS),Lipoteichoic acid,Bacterial lipopeptides,Peptidoglycan,Yeast and gram,+,bacteria,Bacterial DNA(CpG),Flagellin,Terminal mannose/fucose,Viral DNA(CpG),ssRNA,dsRNA,P,athogen-,A,ssociated,M,olecular,P,atterns,(PAMP),是病原微生物（尤其是原核生物）表面存在一些人体所没有的，但可为许多相关微生物所共享、结构恒定、进化保守的分子结构。,损伤相关分子模式,（,damage-associated molecular patterns,，,DAMPs,）,机体自身细胞所释放的内源性分子，即内源性危险信号，来源于受损或坏死组织和某些激活的免疫细胞。主要有,HMGB1,、热体克蛋白等。,PAMP vs DAMP,Sterile inflammation,conserved microbial motifs VS,non-microbial signals,Toll-like Receptors,Locations of Different PRRs,Body fluids,-Soluble PRRs,Cellular PRRs,-Cell surface,-Endosomes,-Cytosol,Surface receptors of macrophages,PRR,甘露聚糖凝集素（,MBL,）,C,反应蛋白（,CRP,）,脂多糖结合蛋白（,LBP,）,可溶性：体液和血液,细胞吞噬型：细胞膜,甘露糖受体（,MR,）,清道夫受体（,SR,）,补体受体（,CR,）,Fc,受体（,FcR,）,甲酰甲硫氨酰肽受体（,fMLP,）,信号转导型,细胞膜,内体、溶酶体,细胞质,TLR1,、,2,、,4,、,5,、,6,、,10,、,11,TLR3,、,7,、,8,、,9,NLRs,、,RLRsALRs,固有免疫细胞表面、内体、溶酶体、细胞质中、可识别一种或多种,PAMPs,或,DAMPs,的识别分子。,Diversity of pattern recognition receptors,Toll-Like Receptors(TLRs),MyD88-Dependent and independent Signaling,NLRs are cytoplasmic bacterial sensors that activate inflammasomes,1,Viral Pattern Recognition Receptors:Signaling and Consequences,Interaction between innate and&adaptive immunity,1.Innate immunity=Ag presentation(by Dendritic cells),2.Adaptive immunity=Ag recognition(by T&B lymphocytes),适应性免疫,（,adaptive immunity,）,是机体获得性、抗原特异性、抵抗病原微生物感染的高效防御机制。,获得性免疫（,acquired immunity),或特异性免疫,(specific immunity),，是个体出生后，在环境中受抗原刺激所产生的免疫力，针对特定抗原，有特异性、多样性、记忆性和耐受性。,1),特异性，对某个特定的异物性抗原能引起特异性免疫应答；指抗原特异性。,2),多样性，机体可针对环境中多种多样的抗原，分别建立起不同的特异性免疫应答；多样性是特异性产生的基础。,3),记忆性，当异物抗原再次入侵时，可产生快而强的再次免疫应答效应；记忆性淋巴细胞,。,4),耐受性，正常情况下，免疫系统对自身成分有保护性的免疫耐受；,抗原决定簇,Antigenic determinant,，,AD,抗原分子表面具有特殊立体构型和免疫活性的化学基团称为,抗原决定簇或抗原决定基,。,由于抗原决定簇通常位于抗原分子表面，因而又称为,抗原表位,(,epitope,),。,抗原决定簇,抗原决定基,抗原表位,抗原决定簇决定抗原的特异性，即决定抗原与抗体发生特异性结合的能力,（实际是抗原,决定簇与抗体的结合,）。,AD,的数目、性质和空间构象决定抗原特异性,抗原以,AD,与相应抗原受体及抗体特异性结合,T,细胞表位和,B,细胞表位,T,细胞表位,：,TCR,识别,必须经降解加工处理后才能被,T,细胞识别,线性决定簇,B,细胞表位,：,BCR,识别或抗体识别并结合,直接识别,构象决定簇或线性决定簇,抗原提呈细胞,(antigen-presenting cell,APC),是指能摄取,加工处理抗原,并将抗原信息提呈给淋巴细胞的一类免疫细胞，在机体免疫应答过程中发挥重要作用。,此类细胞能辅助和调节,T,细胞、,B,细胞识别抗原并对其产生应答，故又称为辅佐细胞,(accessory cell),，简称,A,细胞。,APC,加工处理的抗原种类,:,外源性抗原,(exogenous antigen):,通过吞噬或吞饮等作用被,APC,从细胞外摄入的抗原，以抗原肽,-MHC I I,类分子复合物形式提呈给,CD4,+,T,细胞。,内源性抗原,(endogenous antigen):,细胞内合成的抗原，以抗原肽,-MHC I,类分子复合物形式提呈给,CD8,+,T,细胞。,外源性抗原加工，处理及提呈,APC,摄取的外源性抗原在内体中降解成肽，与,MHC,类分子,(,在内质网合成,),结合后表达于细胞表面。外源性抗原加工中需要,Ii,链和,HLA-DM,分子的参与。,Ii,链 与,MHC,类分子的转运有关，并通过,CLIP,封闭,MHC,类分子的肽结合部位，阻止,MHC-,类分子在内质网中与内源性抗原肽结合。,HLA-DM,分子促使,CLIP,从,MHC,类分子肽结合区解离，有利抗原肽与,MHC,类分子结合。,内源性抗原加工，处理及提呈,内源性抗原经蛋白酶体降解成肽，通过抗原加工相关转运体,(TAP1,、,TAP2),转运进入内质网，与,MHC,类分子,(,在内质网合成,),结合成肽,-MHCI,类复合物，通过高尔基体表达于细胞表面。,TAP,是内质网上的异源性二聚体，由,TAP,-1,及,TAP,-2,基因编码胞浆中蛋白酶体（,proteasomes,核心成分为低分子量多肽,LMP,细胞被病毒感染后出现的病毒蛋白，基因突变后产生的肿瘤抗原,脂类抗原的,CD1,分子提呈,CD1,分子：非经典,MHC I,类分子,提呈抗原：提呈糖脂或脂类抗原，供,CD1,限制性,T,细胞识别,CD1,分子提呈微生物的,脂质抗原，,脂质抗原由,CD1,分子提呈后激活,CD1,限制性,T,细胞。,Adaptive immunity,Main effectors:,Antibody,T cell receptors,B,BCR,T,TCR,THE ADAPTIVE IMMUNE RESPONSE,Antibody-Mediated Immunity(AMI),Involves B lymphocytes,plasma cells and antibodies,Humoral immunity,Name derives from antibodies found in body fluids(humors-old medical term),Cell-Mediated Immunity(CMI),Involves T lymphocytes,antigen-presenting cells and MHC(major histocompatibility complex)molecules,Cellular immunity,Types of adaptive immunity,1.Humoral immunity,=Molecules in body fluid,e.g.Antibody(Ab),=Key player=,B cells,=Target extracellular,microbes&toxins,2.Cell-mediated immunity,=Key player=,T cells,=,regulate other immune,cells,=Target intracellular,microbes,e.g.viruses,bacteria,For innate immunity,it also includes,Humoral,&,Cellular,components for immune defense,1,、抗原提呈与识别阶段（感应阶段）：,2,、活化、增殖、分化阶段（反应阶段）：,T,细胞活化、增殖分化为效应,T,细胞；,B,细胞活化、增殖分化为浆细胞；,部分细胞发育为记忆细胞。,3,、效应阶段：,效应,T,细胞对抗原的清除；,浆细胞分泌抗体清除抗原。,免疫应答的三个阶段,Overview of adaptive immune responses,CELL-MEDIATED IMMUNITYCMI),Directed against intracellular microorganisms,Non-phagocytic cells and phagocytic cells,T-lymphocytes(T cells),Differentiate into effector cells following antigen presentation by antigen presenting cells(APCs),Functional types of T cells,Helper(CD4 T cells),TH1 and TH2 cells,Cytotoxic(CD8 T cells),T cells develop in the thymus,T cells develop in bone marrow,but in order to mature,they need to migrate to thymus=,thymus-(T)-dependent lymphocytes,Thymus is a primary lymphoid organ located in the,upper thorax,(chest),just above the heart.,Progenitor(precursor)T cells enter the thymus,where they mature(=produce T cell receptors).Mature T cells leave the thymus and enter the secondary lymphoid tissues,where they become activated after exposure to antigen.,T cells develop in bone marrow and then migrate to thymus where they mature,Mature T cells enter the blood stream,and after infection accumulate in the secondary lymphoid tissues where they are activated.,T cell development,TSC,CD4 RTE,DN,Pre-TCR,DP,TCR,CD4,CD8,TCR,TCR,CD8,CD4 SP,TCR,CD4,CD4,b,-selection,Positive,selection,Negative selection,Functional maturation,TCR-,b,rearrangement,TCR-,a,rearrangement,TCR,CD8,CD8 SP,CD8 RTE,Development of Thymocytes,Double,negative,Double,positive,Single,positive,Notch Signal and T-lineage Commitment,受体,:,Notch 1-4,；胸腺细胞表达,Notch1-3,；全部效应由,Notch 1,介导,配体：,Dll1,，,3,，,4,，,Jagged 1,，,2,；胸腺上皮细胞表达全部配体；,Dll4,可能为生理性配体,T,系定向：,始于,DN1,，完成于,DN3,；绝对依赖,Notch1,信号,T cells undergo further differentiation in secondary lymphoid tissues after encounter with antigen,Only a small fraction of,naive T cells,(mature T cells before they encounter antigen)survives the positive and negative selection,and leaves the thymus.,Mature naive T cells can re-circulate between blood and lymphoid tissues for many years(in contrast to B cells,which have shorter life span).,In secondary lymphoid tissues,T cells accumulate in,T cell areas,where they become activated by their specific antigens.,Encounter with antigen induces the final stage of T cell development:their differentiation into,effector T cells,.Some effector T cells stay in the lymphoid tissues(,CD4-TH2 cells,),while others migrate to site of infection(,CD8 and CD4-TH1 cells,).,T,细胞受体复合物,由,TCR,和,CD3,组成。前者识别和结合抗原肽,后者将,TCR,获得的抗原信号传递至细胞内。,T,细胞对抗原的识别,APC,T,细胞,CD molecules associated with reco-gnition,adhesion and activation of T cell,MHC-,MHC-,B7,CD40,CD58,CD2,信 号,第一信号,第二信号,T,细胞,TCR,和,CD4/CD8,CD28,APC,MHC-,肽,复合物,B7,(B7.1,、,B7.2),T,细胞活化的双信号刺激,第一信号：,TCR,对,MHCII,抗原肽复合物的识别，,CD3,分子将第一信号传递到细胞内。第二信号：,CD28,识别专职,APC,上的,B7,分子,又称协同刺激信号。,Effector T cells,In contrast to terminally differentiated B cells(plasma cells),there are several types of terminally differentiated effector T cells.,CD8 T cells Cytotoxic T cells,(recognize MHC class I molecules),CD4 T cells,T,H,1 helper cells,(activate macrophages),T,H,2 helper cells,(induce differentiation of B cells into plasma cells and production of antibodies),Activation(cytokines),(recognize MHC II molecules),The immune system is maintained in a carefully regulated balance between the two polarised control arms,Th1(cellular immunity)and Th2(humoral immunity).,In disease states the balance is skewed.,multiple sclerosis,rheumatoid arthritis and type I diabetes,have a Th1 bias,whereas cancer patients have a Th2 bias.,Th1 and Th2 Cells,Do not Represent All CD4,+,Cells,More,T helper subsets,Th,3,:,TGF-producing CD4 T cells,Tr,1,:,IL-10-producing CD4 T cells,Th,9,:,IL-9-producing CD4 T cells,T,fh,:,follicular helper T cells,located in the follicular regions of lymph nodes and spleen,follicular Th1/Th2/Th17 cells,ANTIBODY-MEDIATED(HUMORAL)IMMUNITY,Directed against extracellular microorganisms and toxins,B-lymphocytes(B cells),Differentiate into plasma cells which produce antibodies,Function as antigen-presenting cells(APCs),Classification of Antibodies(Immunoglobulins),Immunoglobulin M(IgM),Immunoglobulin G(IgG),Immunoglobulin A(IgA),Immunoglobulin D(IgD),Immunoglobulin E(IgE),Paul Ehrlich:,One of the fathers of humoral,adaptive immunity,1.The Discovery of,Antibody functions,2.The Nobel Laureate in Medicine 1908,Adopted from Nature Immunology,July 2008,抗体的功能,V,区的功能,识别并特异性结合抗原,单体（,IgG,IgE)2,价,二聚体,分泌型,IgA)4,价,五聚体,(IgM)10,价,中和效应,中和毒素和病毒,与,Ag,结合,促吞噬细胞吞噬,抗体的结合价,实际意义,C,区的功能,1.,激活补体系统,Ab(,IgM,、,IgG)+Ag,C1q,补体经典途径,IgG4,、,IgA,和,IgE,的凝聚物,补体旁路途径,2.,介导免疫细胞活性,(1),调理作用（,opsonization,）,：,IgG,+,抗原,(,颗粒性,),Fc,R,（单核、巨噬细胞及中性粒细胞）,促吞噬细胞吞噬；,(2),ADCC,：,IgG,+,抗原,(,靶细胞,),Fc,R,（,NK,细胞）,杀伤,靶细胞；,(3),介导超敏反应,：,型,、,型,和型超敏反应。,3.,穿越胎盘和粘膜,Antibody-Dependent Cellular Cytotoxicity(ADCC),Th2,与,B,细胞的相互作用，获得第二信号：协同刺激信号,CD40-CD40L,活化的,Th2,细胞分泌细胞因子及表达,CD40L,，辅助,B,细胞活化,CD79,/,2,第二信号（,Th,细胞信号）,有二种方式,（,1,）,Th,细胞,-B,细胞间接触作用：,CD40L-CD40,等,（,2,）,Th,细胞分泌细胞因子：,IL-4,、,5,、,6,等,胸腺依赖性抗原（,TD-Ag,）,活化的,B,细胞增殖与分化,活化,B,细胞 浆细胞 产生抗体,原始淋巴滤泡分裂增殖，形成生发中心（一周左右）,Specificity,Memory,and Homeostasis of Adaptive Immunity,体液免疫应答一般规律,多克隆抗体,(polyclonal antibody,，,PcAb),：,采用传统的免疫方法，将抗原物质经不同的途径进入动物体内，经数次免疫后采取动物血液，分离出血清，由此获得的抗血清即为多克隆抗体。,用天然的抗原物质免疫动物，刺激多个,B,细胞克隆所获得的免疫血清（含多种特异性抗体）。,单克隆抗体,(Monoclonal Antibody,，,McAb):,由一个,B,细胞分化增殖的子代细胞产生的针对单一抗原决定簇的抗体，称单克隆抗体。,由一个,B,细胞克隆产生。,识别一种抗原表位。,高度均一（结构、特异性）。,杂交瘤技术制备。,基因工程抗体：,利用基因工程技术来制备的抗体分子称为基因工程抗体，是分子水平的抗体。,抗体针对的靶分子,作用机制,治疗疾病,CD3,阻断,T,细胞功能,预防肾移植排斥反应,CD25,阻断,IL-2,受体,预防肾移植排斥反应,CD20(,或偶联核素,),诱导肿瘤细胞凋亡,non-Hidgkins,淋巴瘤和,RA,CD33(,免疫毒素,),诱导肿瘤细胞凋亡,急性髓样白血病,CD52,诱导肿瘤细胞凋亡,慢性,B,淋巴细胞白血病，,T,细胞瘤,Her2(CD340),抑制和杀伤肿瘤细胞,转移性乳腺癌,EGFR,抑制肿瘤血管形成,转移性结肠直肠癌和头颈部肿瘤，非鳞癌、非小细胞肺癌，对化疗反应差的多形性胶质细胞瘤,CD41/CD61(gpIIbIIIa),抑制血小板凝聚,预防冠状动脉血管形成术中,血栓形成,US,和,EU,所批准的治疗性抗体,抗体针对的靶分子,作用机制,治疗疾病,TNF,阻断,TNF,与受体结合,RA,、银屑病性关节炎、,克隆氏病、强直性脊椎炎,CD11a,抑制白细胞黏附,斑状牛皮癣,VEGF,抑制血管形成,年龄相关性黄斑变性,4,整合蛋白,抑制白细胞黏附,多发性硬化症,IgE,阻断,IgE,与,IgE,受体结合，抑制肥大细胞、,嗜碱性粒细胞释放介质,持续性哮喘,RSVgpF,中和病毒,预防儿童高危期,RSV,感染,US,和,EU,所批准的治疗性抗体,鼠源性单克隆抗体将逐渐被人源化抗体所替代：鼠源性单克隆抗体与人补体成分结合能力低，,CDC,作用相应较弱，对肿瘤细胞的杀伤能力较弱；它与,NK,等免疫细胞表面,Fc,受体亲和力弱，介导的,ADCC,作用较弱；鼠源抗体在人血循环中的半衰期短，它发挥,ADCC,与,CDC,作用的时间较短；鼠单克隆抗体具有免疫原性，宿主易产生抗抗体引起过敏反应。,抗体人源化改造及人源抗体制备,人,-,鼠嵌合抗体,：,应用基因工程技术将小鼠单克隆抗体的恒定区用人源抗体恒定区代替而拼接成嵌合抗体。,改型抗体如,CDR,移植、,SDR,移植,用鼠单克隆抗体的,CDR,、,SDR,移植到人源抗体可变区，替代人源抗体,CDR,、,SDR,。,表面氨基酸残基人源化,抗体人源化的主要技术,提高抗体,效应功能,偶联细胞毒物质,双特异性抗体,抗体,Fc,突变,改变抗体糖基化,细胞内抗体,抗体融合蛋白,提高抗体效应功能,Mj,macrophage,IL-8,Activated,T cell,a,A,D,P56,B,C58,B,C,NH,2,COOH,IL-2,Cytockines are low-molecular-weight regulatory,proteins,or,glycoproteins,secreted by white blood cells and various cells(vascular endothelial cell,epidermic cell and fibroblast)in body in response to a number of,stimuli.,Cytokine,Biological effects,IL-1,TNF,GM-CSF,M-CSF,FGF,PDGF,VEGF,IL-12,IL-15,IL-6,LIF,OSM,Chemokines,TGF,IL-10,IL-11,IL-13,sTNF-R,IL-1ra,PRO-INFLAMMATORY,ANTI-INFLAMMATORY,Cytokine imbalance during inflammation,细胞因子的研究热点,1,、新细胞因子的基因克隆化,2,、细胞因子受体的基因克隆化,3,、细胞因子信号转导机制,4,、新一代细胞因子：高活性，多功能，低毒副作用，长半衰期，高稳定性,5,、细胞因子作为生物应答调节剂（,BRM,）的临床应用,6,、细胞因子表达调控,7,、细胞因子基因治疗,Infections occur when the physical barriers of epithelium are breached,or when pathogens adhere to the epithelium,Pathogen replication,Recruitment of neutrophils and macrophages,Inflammation,Pathogen adherence to epithelium,Pathogens are carried to the closest lymph node where they stimulate specific T-and B-cells,Production of specific antibodies=,Adaptive immunity,Innate immunity,Adaptive immunity,Activation of adaptive immunity by innate immunity,Time course of primary infection in healthy individuals,and people who lack immune or adaptive immunity,In the absence of innate immunity,there is also no adaptive immunity,Infection is cleared from the body by combined actions of innate and adaptive immunity,T Ce