博士生课程固有免疫模式识别ppt课件.ppt

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1、Innate Immunity,monocyte,macrophage,bacteria,The most ancient defensePhysical&chemical barriers and cellular lineRecognition by the innate immune system sets the stage for an effective adaptive immune response.,机体在种系发生和进化过程中逐渐形成的一种天然免疫防御功能，构成机体抵御病原生物入侵的第一道防线.,复习,一、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-

2、脑屏障、血-胸腺屏障血-胎屏障、气-血屏障,单核-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、,Physical,chemical and microbiological barriers of our body,1、固有免疫屏障,This may cause inflammation and bleeding,Normal Flora competing with Invading Pathogens.Antibiotic treatments disrupt th

3、e natural ecology of the colon,2、固有免疫细胞,PhagocyteNKILLs（固有样淋巴细胞）DC MCBasophil Eosinophil,T细胞 NKT细胞 B1细胞,Monocyte-macrophageNeutrophil,Recognition of an infection once it gets past the epithelial barrier,Polarization of Tumor-associated macrophages(TAM),分泌,IL-1,IL-6,IL-12,TNF-,a,IL-8,GM-CSF,细胞因子,酶,其它

4、因子,杀伤,肿瘤细胞,抗原,呈递作用,前列腺素,白三烯,补体成分,纤维蛋白,结合蛋白,凝血因子,溶菌酶,酸性水解酶,赖氨酸酶,酯酶,胶原蛋白酶,弹性纤维蛋白酶,免疫调节作用,吞噬并杀伤,病原微生物,巨噬细胞的功能,Figure 8-19 part 2 of 2,Leukocyte recruitment to sites of infection:a multi-step navigation,1.Selectins2.Chemokines3.Integrins,IL-8,Interaction between Neutrophils and Endothelium,Cellular Adhe

5、sion Molecules(CAMs):Mucin-like CAMsSelectinsIntegrinsIg-superfamily CAMs,Killer activatory receptor,Killer inhibitory receptor,KIR:KIR2DS,KIR3DS KLR:CD94/NKG2C NKG2D NKp46 NKp30 NKp44,NCR,KIR2DL,KIR3DLCD94/NKG2A,Bind class I HLAmolecules,Function,Bind non-class I HLA molecules,Receptors associated

6、with killer activation and killer inhibition on NK cells,2、固有免疫细胞,PhagocyteNKILLs（固有样淋巴细胞）DC MCBasophil Eosinophil,T细胞 NKT细胞 B1细胞,Monocyte-macrophageNeutrophil,过敏性疾病,抗原的处理与提呈,Nuocytes represent a new innate effector leukocyte that mediates type-2 immunity.Nature.2010 Mar 3.,Type-2 immunity:responsib

7、le for protective immune responses to helminth parasites and the underlying cause of the pathogenesis of allergic asthma.Type-2 cytokines:interleukin IL-4,IL-5 and IL-13.Nuocytes expand in vivo in response to the type-2-inducing cytokines IL-25 and IL-33,and represent the predominant early source of

8、 IL-13 during helminth infection.In the combined absence of IL-25 and IL-33 signalling,nuocytes fail to expand,resulting in a severe defect in worm expulsion that is rescued by the adoptive transfer of in vitro cultured wild-type,but not IL-13-deficient,nuocytes.,3、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

9、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,补体系统,细胞因子和免疫相关细胞表面分子,二、固有免疫识别,病原相关分子模式(Pathogen-associated molecular patterns,PAMPs)损伤相关分子模式（damage-associated molecular patterns，DAMPs）模式识别受体（Pattern Recognition

10、Receptors）,病原相关分子模式(Pathogen-associated molecular patterns,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。PAMP的特征 1.通常为病原微生物所特有,乃天然免疫系统区分“自己”与“非己（微生物）”的分子基础。脂多糖：多数革兰阴性菌细胞壁成分；磷壁酸：多数革兰阳性菌胞壁成分；肽聚糖：革兰阳性/阴性菌、真菌胞壁成分；甘露糖：微生物细胞壁上糖蛋白和糖脂成分2.为微生物生存和致病性所必需 PAMP突变或缺失 微生物死亡或微生物对外界环境适应性3.宿主泛特异性识别的分

11、子基础 PAMP是由一群或一类特定的微生物所共有的恒定结构（如LPS）。宿主由种系编码的有限数量PRR 可察觉任何微生物感染的存在,Pathogen-AssociatedMolecularPatterns(PAMP),Innate immune recognition of bacterial cell wall components,Gram-negative bacteria,Gram-positive bacteria,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

12、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,模式识别受体（Pattern Recognition Receptors，PRRs）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。,PRR,甘露聚糖凝集素（MBL）C反应蛋白（CRP）血清淀粉样蛋白(SAP)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（

13、CR）Fc受体（FcR）甲酰甲硫氨酰肽受体（fMLPR）,信号转导型,细胞膜内体、溶酶体细胞质,TLR1、2、4、5、6、10、11、12、13,TLR3、7、8、9,NLRs、RLRs、ALRs,EXTRACELLULAR/SECRETED PRRs,Mannose binding lectin/protein(MBP)C-reactive protein(CRP)Serum amyloid protein(SAP)LPS-binding protein(LBP),Acute phase proteins,Acute phase response(APR):the serum changes

14、APR proteins:their concentrations rose or fell,(during the acute phase),Sites of injury or infection signals(proinflammatory cytokines:TNF-,IL-1,&IL-6 produced by phagocytes)stimulatingLiver:synthesis of APR proteins Increase in the level of C-reactive protein&Mannose-binding lectin/MBL&Serum amyloi

15、d protein/SAP&fibrinogen),Involved in Clotting,Two Secreted PRRs:CRP,MBP,SAP made in acute phase liver response,Mannose binding lectinLung surfactants A,DFicolins“pattern recognition receptors”;in this case pattern of terminal sugars on cell surfaces,Recognizing mannose-containing molecular patterns

16、 found on microbes but not on vertebrate cells directing complement attack,Mannose-binding lectin,Mannose binding protein(MBP)Part of C-type lectin superfamilyAssociates with and activates serine proteases:MASP-1 and MASP-2 After binding to pathogen surface this complex activates lectin pathway of c

17、omplement system,C2 and C4,MB-LECTINMASP=MBL-associated serine proteaseMBLmannose,MB-LECTIN,ANOTHER VERSION POINTS OUT TOTALITY OF CLEAVED C3 FUNCTIONS,Bind to phorphorylcholine(PC)on bacteria,other microorganisms,damaged host cell membranesPC found in teichoic acids,capsular carbohydrates,and lipop

18、olysaccharides Requires Ca+Function directly as opsonins(enhancer of phagocytosis)Function indirectly by binding to C1q of classical complement pathway and activate complement cascade,C-reactive protein(CRP)and serum amyloid protein(SAP),(belongs to a family of pentameric protein called pentraxins)b

19、inding to polysaccharide&phophorylcholine(=ligands)on the cell wall of bacteria&fungi in a calcium-dependent reaction activating complement system lysis,opsonization promoting phagocytosis&pathogen clearance,Lipid transfer molecule binds to monomeric LPS and to high-affinity LPS receptor named CD14

20、and,on macrophage,neutrophils,DCsLBP+bactericidal permeability increasing protein(BPI)binds LPS on bacteria and then to CD14,a high affinity LPS receptor.See later TLR4,LPS-binding protein(LBP),Extracellular factor(LPS)carried by LBP to CD14 where it binds to TLR4 and then MD2 binds,Simplified Versi

21、on,模式识别受体（Pattern Recognition Receptors，PRRs）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。,PRR,甘露聚糖凝集素（MBL）C反应蛋白（CRP）血清淀粉样蛋白(SAP)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（fMLPR）,信号转导型,细胞膜内体、溶酶体细胞质,TLR1、2、4、5、6、10、11、12、13,TLR3、7、8、9,NLRs、RLRs、ALRs,MANNOSE RECEPTO

22、RThe mannose receptor(MR)is a 175 kDa type I membrane molecule expressed in the mouse by most tissue macrophages and lymphatic and hepatic endothelia.,Glycoprotein PRRs recognizeLPS and lipoteichoic acidIntact G-and G+bacteriaDamaged host cells and tissuesApoptotic and senescent cellsmodified low-de

23、nsity lipoproteinsSix classes,Scavenger receptors,甲酰甲硫氨酰肽受体（fMLPR）,Staphylococcal Protein A Inhibits Phagocytosis by Blocking Fc,果蝇的Toll受体胞浆的功能域与IL-1受体很相像(Toll/IL-1 receptor(TIR)domain)，显然具有重要的免疫功能。Toll突变后果蝇很容易受霉菌感染。Cell 86:973-83.,Toll-like receptor(TLR),Julie A.Hoffmann,Ph.D.Strasbourg,France,In 1

24、996,Hoffmanns group Toll functions as a PRR in Drosophila,Toll-Like Receptors(TLRs)Total of 13 TLRs have been identified in mammalsHuman(TLRs 1-10)Mouse(TLRs 1-9,11-13)In general TLRs recognize constituents of microbial cell walls or pathogen-specific nucleic acids that are essential to the integrit

25、y,function or replication of microbes/viruses that cannot readily be modified.,Toll-Like Receptors(TLRs),There are six major clades of TLRs,each recognizing a general class of molecular patterns.,Molecular tree of the vertebrate Toll-like receptors(TLRs).,The evolution of vertebrate Toll-like recept

26、ors,Toll-Like Receptors(TLRs):Basic Architecture,Structural organization of human TLRs.,ENDOTOXIN,-Structural component of the outer leaflet of the outer membrane of Gram negative bacteria-Contains the O-antigenic polysaccharide determinant.-Presence of polysaccharide and lipid components.-Also term

27、ed Lipopolysaccharide(LPS).-The lipid component or Lipid A(glycophospholipid)is responsible for the biological activities of LPS(toxicity/fever),LPS(endotoxin),Outer membrane,Peptidoglycan,Inner membrane,O-specific chain Core Region Lipid A,LPS,the toxic center of LPS,The Basic Architecture of LPS S

28、tructure of Lipid A,Lipid A is a glycophospholipid withphosphorylated D-glucosamines,LBP and CD14 the first Endotoxin Receptors,LPS-Binding Protein(LBP)-Plasma protein produced in the Liver.-Delivers LPS from the serum to macrophages.-Enhances the sensitivity of macrophages for LPS.CD14-GPI-linked m

29、embrane protein(macrophages).-Also exists in a soluble form recruited to endothelial cells.-Works with LBP to bring LPS to the cell surface.No link between LBP/CD14 and intracellular signaling.However,LPS detection by immune cells results in an intracellular signaling response and production of TNF.

30、,Delivery of LPS to TLR4 by lipid transfer proteins,BiologicalActivities,LBP-CD14Co-receptor(s),MD-2,TLR4,TLR signaling pathways,The generation of inflammatory cytokines and chemokines The generation of antimicrobial peptides,Initiation,Signal-induced assembly of pathway components/involvement of an

31、 adaptor molecule,Protein kinase-mediated phosphorylation,Initiation of an enzyme cascade:MAP kinase pathway(in many cell types)NFB pathway(a powerful transcriptional factor),generate cytokines,adhesion molecules&other effectors,affectthe cell cycle or cellular differentiation,Conserved pathways in

32、innate immunity in Drosophila and mammals.,Other Bacterial Poisons,Lipopeptides,Peptidoglycan,Lipoteichoic Acid,Unmethylated DNA Oligomers,TLR4 LigandsLipopolysaccharides two grades of purity:-Standard LPS:LPS-EB and LPS-EK are standard preparations of lipopolysaccharide.They are extracted by a phen

33、ol-water mixture.LPS-EB and LPS-EK contain other bacterial components,such as lipopeptides,and therefore stimulate both TLR4 and TLR2.-Ultra-pure LPS:Ultrapure LPS-EB are extracted by successive enzymatic hydrolysis steps and purified by the phenol-TEA-DOC extraction,TLR5 LigandsFlagellin,TLR6/TLR2

34、Ligand-MALP-2,TLR2 Ligands-Heat-Killed Bacteria-Lipoglycans-Lipopolysaccharide-Lipoteichoic Acids-Peptidoglycans-Synthetic Lipoproteins-Zymosan,TLR3 LigandsPoly(I:C)a synthetic analog of double-stranded RNA(dsRNA)Poly(A:U),TLR9 Agonists Stimulatory ODNs-CpG ODNs-Control ODNs-Labeled ODNs E.coli DNA-

35、E.coli DNA ef-E.coli ssDNA,TLR7 Ligands(human&mouse TLR7)-CL264:Adenine analog-Gardiquimod:imidazoquinoline compound-Gardiquimod-Imiquimod:imidazoquinoline compound-Imiquimod VacciGrade NEW-Loxoribine:guanosine analogueTLR8 Ligands(human TLR8&mouse TLR7)-Single-stranded RNAs-E.coli RNA TLR7/8 Ligand

36、s(human,mouse TLR7&human TLR8)-CL075:thiazoloquinoline compound-CL097:water-soluble R848,imidazoquinoline compound-Poly(dT):thymidine homopolymer phosphorothioate ODN-R848:Imidazoquinoline compound,MyD88-Dependent and independent Signaling,TLR4 can signal using both pathways,TRAM=TRIF-related adapto

37、r moleculeTIRAP=Toll-interleukin 1 receptor(TIR)domain-containing proteinTLR4 signaling induces the transcription of both proinflammatory cytokines and IFN-that are required for anti-bacterial and anti-viral immune responses,respectively.,Cooperation of Toll-like receptor signals in innate immune de

38、fense,Negative regulation of Toll-like receptor 4(TLR4)signaling,The negative regulators of TLR signaling pathways,NLRs are cytoplasmic bacterial sensors that activate inflammasomes,IL-1 family members,Common sources of interleukin-1(IL-1)family cytokines,ICE:IL-1-converting enzyme,caspase 1,Pro-IL-

39、18:24 kDa and mature IL-18:18 kDaPro-IL-1beta:33 kDa and mature IL-1beta:17 kDapro-caspase-1:45-kDa mature caspase-1:20KD 10KD,Inflammasomes are molecular platforms activated upon cellular infection or stress that trigger the maturation of proinflammatory cytokines such as IL-1b and IL-18 to engage

40、innate immune defenses.,THE INFLAMMASOMES,Activators of the inammasome.,The Human NLR Family Members,Human and Mouse NLR Family Members,DOMAIN ORGANIZATION OF REPRESENTATIVE NLRs,The NLRs family is composed of 20 members.They contain:a C-terminal leucine-rich repeat domain,LRR a central nucleotide-b

41、inding domain,NACHT an N-terminal protein-protein interaction domain composed of CARD(caspase activation and recruitment domain)Pyrin domain or Bir(baculovirus inhibitor of apoptosis repeat)domain,Schroder and Tschopp Cell 2010,NLRP1,NLRP3,IPAF,and AIM2 Inammasomes,What is an inflammasome and what d

42、oes it do?,Activation models and composition of NLRP3 inflammasome,Schroder and Tschopp Cell 2010,PAMPs:MDP,LPS,viral/bacterial RNA,aerolysin,etc.DAMPs:ATP,hyaluronan,uric acid,amyloid-b peptide,ROS,silica,asbestos,UVB,etc.,The channel model of NLRP3 inflammasome activation.,Proposed pathways for NL

43、RP3 activation.,A role for mitochondria in NLRP3 inflammasome activation,Pathways for activation of the inflammasome by PAMPs and DAMPs,The AIM2 inammasome,The role of the inflammasomes in human disease,The influence of inflammasome activation on adaptive immunity,RLRs 等病毒识别受体Host Response to Virus

44、Infection,Viral Nucleic Acids double stranded(ds)RNA single stranded(ss)RNA DNA(hypomethylated or CpG rich)“Some”Viral Proteins-uncommone.g.glycoproteins of Herpesvirus and Respiratory Syncytial Virus,Viral Pathogen-Associated Molecular Patterns(PAMP),Production of Type 1()interferons Production of

45、inflammatory mediators:cytokines and chemokines Mechanism:Interferon Response Factor 3/7(IRF3/7)activation NFB activationTranslocation from cytoplasm to nucleus,Engagement of Viral PAMP Receptors on/in Cells,cell surface and endosomal localizationcell cytoplasm localizationextracellular space locali

46、zation,Viral PAMP Receptors=anti-viral sensors=anti-viral PRR,Viral PAMP Receptors(anti-viral sensors),TLR-Toll Like Receptors:cell surface&/or endosome Endosomal TLR TLR-9 CpG DNA(methylated viral DNA)TLR-7/8 ssRNA TLR-3 dsRNA TLR 9 and 7 are major triggers of Type 1 IFN production-particularly by

47、plasmacytoid DC:Cell surface TLR TLR-2 and TLR-4:viral glycoproteins,HCV,RSV,The third member of the RLR family,LGP2,lacks any CARDs and was originally identified as a negative regulator of RLR signaling.Nevertheless,LGP2 and its ATPase activity were dispensable for the responses to synthetic RNA li

48、gands for MDA5 and RIG-I.LGP2 facilitates viral RNA recognition by RIG-I and MDA5 through its ATPase domain.,PNAS January 26,2010 vol.107 no.4 1512-1517,Virus recognized by NLRs.,Viral PAMP Viral RNA,DNA,Protein,Viral PAMP R TLR,RLR,NLR,Signalling,Nuclear Factor B(p50p65)+Interferon Response Factor

49、3/7,Activation/Translocation to Nucleus,Type 1 Interferon 300+Anti-viral Genes,Cytokines/Chemokines Inflammatory Response,1,Viral Pattern Recognition Receptors:Signaling and Consequences,Response of cells to interferons,Anti-viral effects of interferon a/b,活化的PKR能够识别蛋白质合成的起始因子真核翻译起始因子-2(eukaryotic i

50、nitiation factor-2，eIF-2)，催化eIF-2a 51位苏氨酸磷酸化，改变eIF2功能，阻止eIF2B发挥作用，从而降低eIF2-GTP的水平，使蛋白合成和病毒复制水平下降,Viruses have evolved mechanisms/proteins that can block TLR and/or RLR signaling at different steps in the signaling pathway,the end result is to supress the anti-viral(IFN)response and the induction of