免疫學track課件天然免疫

1、序號教師單位授課教師授課日期授課內(nèi)容1北京大學蔣爭凡4.30天然免疫及其相關細胞信號轉導2上海巴斯德所李斌5.7獲得性免疫及免疫調(diào)節(jié)3上海巴斯德所李斌5.8代謝與免疫4北京大學醫(yī)學部張毓5.14淋巴細胞發(fā)育I5北京大學醫(yī)學部張毓5.15淋巴細胞發(fā)育II6中國科學院生物物理所秦志海5.21腫瘤免疫I7中國科學院生物物理所秦志海5.22腫瘤免疫II8中科院微生物研究所方敏5.28NK細胞發(fā)育與功能I9中科院微生物研究所方敏5.29NK細胞發(fā)育與功能II10北京大學醫(yī)學部韓文玲6.4細胞因子11北京大學醫(yī)學部韓文玲6.5免疫細胞膜分子12浙江大學魯林榮6.11B細胞發(fā)育及其功能13浙江大學魯林榮6.

2、12免疫耐受與自身免疫病14暨南大學尹芝南6.18T細胞免疫學15暨南大學尹芝南6.19腫瘤免疫學2014-2015學年第二學期（春季）-免疫學track課程表助教：張潔 手機： 郵箱： 天然免疫及其細胞信號轉導蔣爭凡 北京大學生命科學學院天然（固有）免疫獲得性免疫（T、B細胞）病原微生物感染An immune system must do three things:Detect infection.Eliminate or contain infection.Be tolerant to self.The question “how do we detect infection?” turn

3、s out to be closely related to the question, “how do microbes harm us?”Infections come in all shapes and sizes, and over the history of our species, have killed more human beings than anything else.Staphylococcus aureusNeisseria meningitidisHerpes simplexVariola major(smallpox)MeaslesWhat we die of,

4、 and how long we live, depends very much on when and where we live.Courtesy J-L Casanova59 million people die eachyear of all causes, worldwide.infection accounts for about a quarter of all deaths.Infection has been a strong selective pressure throughout the evolution of our species and throughout t

5、he evolution of multicellular organismsInfection impelled the evolution of our immune systems: both innate and adaptive.Many genes have evolved or have been co-opted to serve immunity.This means that there is much that can go wrong with immunity, and many mutations lead to immunodeficiency or to aut

6、oimmunity.Major causes of death inVictorian England (1837-1901)SmallpoxTuberculosisTyphoidCholeraSarah NelmesJames PhippsEdwardJennerOn 14 May 1796, Jenner tested his hypothesis by inoculating James Phipps, an eight-year-old boy who was the son of Sarah. He scraped pus from cowpox blisters on the ha

7、nds of Sarah Nelmes, a milkmaid who had caught cowpox. This inoculation produced in Phipps a fever and some uneasiness, but no full-blown infection. Jenners Hypothesis:The cowpox, transformed from smallpox, protected milkmaids from the lethal infection of smallpox .Observation: milkmaids were genera

8、lly immune to smallpox. Jenners unique contribution was not that he inoculated person with cowpox, but that he then proved that they were immune to smallpox. Moreover, he demonstrated that the protective cowpox pus could be effectively inoculated from person to person, not just directly from cattle.

9、 Edward Jenner, FRS; (1749 1823)An English physician and scientist who was the pioneer of smallpox vaccine, the worlds first vaccine. He is often called the father of immunology, and his work is said to have saved more lives than the work of any other humanAll approaches to infection, up to this tim

10、e took place in intellectual darknessThe link between infection and microbes had yet to be made.Until it was made, there were simply vague ideas about how infections spread between individuals, and how they produced effects akin to poisoning.Miasma Contagion vs.Generally speaking, infectionresembled

11、 putrefaction oforganic materials (decayingmeat or plant matter) with theproduction of foul smellinggases such as hydrogensulfide, ammonia, mercaptans Putrescent materials of plant and animal origin are toxic when injected into animals, eliciting feverAlbrecht von Haller (1708-1777)Franois Magendie

12、(1783-1855)瑞士解剖學家、生理學家法國生理學家Attempts to purify the “putrid poison”Ernst von Bergmann(1836-1906)Theodor Billroth(1829-1894)Peter Panum(1820-1885)奧地利醫(yī)生，“外科之父”德國醫(yī)生，發(fā)明壓力蒸汽滅菌法Joseph Lister(1827-1912)英國醫(yī)生無菌術之父，發(fā)明消毒法1860無菌可吸收縫線1867The putrid poison is not volatile and is not a known simple end product of p

13、utrefaction or fermentation. It can be differentiated from living microorganisms, which may be a source but not the cause.The toxin resists heat and, thus, differs from typical enzymes (at that time called in German Fermente).It is insoluble in pure alcohol but soluble in water. The protein-like sub

14、stances frequently present in putrid fluids are not toxic by themselves, but they absorb (“condense”) the toxin on their surface when precipated. The toxic principle can be, at least partially, eluted from the precipitates.Injection of 12 mg of the concentrate suffices to produce high fever and kill

15、 a dog.-Peter Panum, 1874 (writing of his own work, performed in 1856).Peter Panum(1820-1885)丹麥醫(yī)生，描述麻疹流行病的第一人Robert Koch(1843-1910)Louis Pasteur(1822-1895)At the time Panum did his work, he had no idea the “putrid poison” came from microbesFerdinand Cohn (1828 1898)Founders of bacteriology19世紀最有成就的科

16、學家之一。他的研究使整個醫(yī)學邁進了細菌學時代，得到了空前的發(fā)展。美國學者麥克哈特所著的影響人類歷史進程的100名人排行榜中，巴斯德名列第12位，可見其在人類歷史上巨大的影響力。2005年，法國舉行了“最偉大的法國人”的評選活動，巴斯德名列第二位，僅次于夏爾戴高樂。1、發(fā)酵作用是由于微菌的發(fā)展， “巴氏殺菌法”用于殺滅細菌。-自然發(fā)生論的否定2、傳染病都是微菌在生物體內(nèi)的發(fā)展：發(fā)現(xiàn)并根除了一種侵害蠶卵的細菌，拯救了法國的絲綢工業(yè)。3、染病的微菌，在特殊的培養(yǎng)之下可以減輕毒力，使他們從病菌變成防病的藥苗。他意識到許多疾病均由微生物引起，于是建立起了細菌理論。他于1843年發(fā)表的兩篇論文“雙晶現(xiàn)象研

17、究”和“結晶形態(tài)”，開創(chuàng)了對物質光學性質的研究。1856年至1860年，他提出了以微生物代謝活動為基礎的發(fā)酵本質新理論。1857年發(fā)表的“關于乳酸發(fā)酵的記錄”是微生物學界公認的經(jīng)典論文。1880年后成功地研制出雞霍亂疫苗、狂犬病疫苗等多種疫苗，其理論和免疫法引起了醫(yī)學實踐的重大變革。1882年，開始研究狂犬病，證明病原體存在于患獸唾液及神經(jīng)系統(tǒng)中，并制成咸毒活疫苗，成功地幫助人獲得了該病的免疫力，在1889年發(fā)明了狂犬病疫苗。發(fā)展了一項對人進行預防接種的技術。路易斯巴斯德(1822-1895)巴氏殺菌法：6065作短時間加熱處理，殺死有害微生物。羅伯特 科赫(1843-1910) 德國醫(yī)生和細

18、菌學家，世界病原細菌學的奠基人和開拓者主要工作： 1876 科赫明確指出細菌是導致炭疽熱這個疾病的“元兇”。科赫的研究確立了疾病的細菌起源理論。1877 第一次發(fā)明了細菌照相法。細菌干燥固定，甲基藍染色，照相。使用蓋玻片制備永久裝片。 1881 第一次使用無菌的土豆片做為固體培養(yǎng)基，平板技術用于菌落分離細菌的純培養(yǎng)。1882 分離出了結核桿菌 結核分枝桿菌，并且證實結核桿菌是這種傳染病的病因。 1884 科赫提出了他最為著名的理論 “科赫原則”。 1. 在人類與動物體內(nèi)結核損傷的器官中都可以發(fā)現(xiàn)結核桿菌的存在，而健康機體上沒有。 2. 可以從患者在血清中得到結核桿菌純培養(yǎng)。 3. 通過向豚鼠接

19、種結核桿菌可以讓豚鼠染上結核病。4. 在患病豚鼠身上再次能夠分離出結核病菌?？坪沼?1905 年被授予諾貝爾醫(yī)學及生理學獎。Ferdinand Cohn (1828 1898)German biologist, one of the founders of modern bacteriology and microbiology.Blue Green AlgaeCohn was the first to classify algae as plants, and to define what distinguishes them from green plants. His classific

20、ation of bacteria into four groups based on shape (sphericals, short rods, threads, and spirals) is still in use today. The first to show that Bacillus(芽孢桿菌) can change from a vegetative state to an endospore state when subjected to an environment deleterious to the vegetative state.Pfeiffer Phenome

21、non:Heat-killed V. cholerae killed guinea pigs, as did V. cholerae injected into guinea pigs that have been immunized against the microbein both cases, there is no infection per se.Pfeiffer coins the term “endotoxin” to describe the poisonous substance associated with bacteria.For Gram negative bact

22、eria, LPS is the endotoxin霍亂弧菌是革蘭氏陰性菌 ，菌體彎曲呈弧狀或逗點狀，一端有鞭毛?；魜y弧菌，為烈性腸道傳染病，在世界上發(fā)生過幾次大流行。By the end of his life, Richard Pfeiffer had been nominated 33 times to receive the Nobel Prize in Physiology or Medicine.Richard Pfeiffer（1858-1945） German physician and bacteriologistWhat is the endotoxin?Bacterial

23、 Cell (E. coli)Lipopolysaccharide (Endotoxin)OuterMembranePeriplasmInnerMembraneCell Wall OrganisationLipid A StructureLipopolysaccharide (LPS) ArchitecturePPGlcNGlcNPPPKdoKdoKdoHepHepHepNH3+nCore RegionLipid AO-specific ChainPutrid poisonEndotoxinLPSHundreds of people die of endotoxin-induced shock

24、 every day, the result of severe Gram-negative bacterial infections.Endotoxic shock is a severe form of systemic inflammation.What does endotoxin do?Chronic Infectious inflammation can be manifested as wastingin chronic diseases-CachexiaAdipocyteCachectinCachectin: a postulated mediator of wasting (

25、cachexia) in chronic diseaseWhat caused the wasting?Connection between TNF & InflammationTNF affects every cell/tissue/organs during infection & inflammation!How LPS leads to the endotoxicity?Other CellsNOPAFLTKininsO2-EndotoxinOr LPSTNF(And Other Cytokines)Whats the receptor for LPS?What is the inn

26、ate immunity?How fly or drosophila survive in a very dirty environment?No adaptive immunityNo T/B cellsNo NK cellsNo AntibodyIn 1989, he predicted that activation of the adaptive immune response is controlled by the more ancient innate immune system.He proposed a general theory of innate immune reco

27、gnition (PRRs) and suggested the principles of innate control of adaptive immunity. Charles Janeway(1943-2003)One of the leading immunologists, formed many of the concepts that are the basis of immunology today. He made major contributions to our understanding of T lymphocyte biology.Most importantl

28、y, he pioneered the modern studies of innate immunity. These predictions were confirmed & now form the conceptual framework for the current understanding of the innate immune system and the links between innate and adaptive immunityPattern Recognition Receptors, PRRsPathogen-Associated Molecular Pat

29、terns, PAMPsDanger-Associated Molecular Patterns, DAMPsThe ReceptorsThe LigandsJaneways pattern recognition theoryThe Breakthrough為什么果蠅、蒼蠅、蚊子等能在骯臟的環(huán)境中生活而不被感染？長滿煙曲霉菌菌絲的果蠅Toll通路突變體1996, Cell, 86:973The C3H/HeJ Mouse andthe Lps LocusResistant to LPS (Heppner and Weiss, 1965)Hyper-susceptible to authent

30、ic G(-) infections (Obrien, et al., 1980; Svanborg-Eden, et al., 1983)One base-pair substitution: C to AAmino acid: P to H天然免疫基因的突變導致人類對病毒感染的抵抗能力顯著下降，甚至引發(fā)死亡！Bruce A. BeutlerJules A. HoffmannRalph M. Steinman2011 Nobel Laureates1）抗感染的第一道屏障2）屬應激反應，發(fā)生迅速 3) 進化上非常保守所有多細胞生物中使用的分子、通路非常保守天然免疫的特點DMTOLL1 HUMT

31、LR4LU-U73916Innate immunity is very conservedImages courtesy Dr. Pamela RonaldBacterial blight of riceXanthomonas oryzae (Xoo)水稻黃單胞菌Monocots (Rice)Insects (Drosophila)Vertebrates (Mouse)Innate immunity is very conserved固有（天然）免疫病原微生物感染誘導細胞產(chǎn)生炎癥因子，殺死病原微生物并活化獲得性免疫的反應。 固有免疫的失控導致： 1）反應不足：機體反復感染乃至死亡； 2）反應過

32、度：自身免疫病、過敏反應甚至猝死、 多器官慢性炎癥反應。研究固有免疫具有重要的理論及實際意義參與天然免疫反應的細胞1）單核巨噬細胞系統(tǒng)： 血液單核細胞 肝臟枯否細胞（kupffer cell） 結締組織組織細胞 骨組織破骨細胞（osteoclast ） 神經(jīng)系統(tǒng)小膠質細胞（ microglia） 脾臟、淋巴結巨噬細胞 肺肺巨噬細胞或塵細胞（dust cell） 功能：吞噬并殺死病菌，引發(fā)炎癥反應。2）嗜中性粒細胞（ Neutrophils ）：吞噬并殺死病菌3）嗜曙紅細胞（ Eosinophils ）：殺死蠕蟲、寄生蟲4）NK細胞（ Natural killer cells ）：殺死被感染細胞

33、，真菌和寄生蟲，激活巨噬細胞5) 成纖維細胞（Fibroblast） Macrophage ingesting yeastgrowth factorTGF-、 family生長因子chemokines 1) C-X-C/亞族，趨化中性粒細胞，主要成員有: IL-8, IP-10, ENA78等； 2) C-C/亞族，趨化單核細胞，包括: 巨噬細胞炎癥蛋白1 (MIP-1、), Rantes等趨化因子colony- stimulating factor G(粒細胞)-CSF、M(巨噬細胞)-CSF、GM (粒細胞、巨噬細胞) -CSF集落刺激因子tumor necrosis factor，TNF

34、腫瘤壞死因子interferon，IFN干擾素interleukins，Ils（1-37）白細胞介素參與天然免疫反應的細胞因子天然免疫反應的啟動器模式識別受體TLRsTLR3,7/8,9RLRHelicaseCARDDNA/RNANucleic acidsAIM2dsDNAdsRNADAI/ZBP1dsDNAMyd88/TRIFMAVS/VISASting/MITAERISNFBIRF3/7CytokinesInflammationInflammasomeCaspase1IL-1 TLR-dependent RLR-dependent NLR-dependentNODsNLRPsIPAFNLR

35、sDAILrrfip1DDX-41IFI16天然免疫及其細胞信號轉導IL-1RIL-1RAcPIL-18RSIGGIRMyD88TLR1TLR2TLR3TLR4TLR5TLR6TLR7TLR8TLR9 TIRDDIgLRRImmunoglobulin domain sub-groupLeucine-rich repeats sub-groupAdaptor sub-groupTLR/IL-1R superfamilyTollTIRAP/MalInnate immune-recognition receptorsTLR1TLR2TLR3TLR4TLR5TLR6TLR7TLR8TLR9 TIRDD

36、LRRLeucine-rich repeats sub-groupToll-Like ReceptorsTollLigands from bacteriaLPS-TLR4Flagellin-TLR5CpG-DNA-TLR9PAMPs from Gram- BacteriaStructure of Cell Wall from Gram-LTA: lipoteichoic acid-TLR2PGN: peptidoglycan-TLR2Flagellin:-TLR5 (A few Gram+ bacteria)CpG-DNA-TLR9PAMPs from Gram+ BacteriaLigand

37、s from bacteriaPGNStructure of Cell Wall from Gram+Gram+Gram-Gram+Gram-MALP2: 2-kDa macrophage-activating lipopeptide from Mycobacterium fermentansLipomannan: from Mycobacterium tuberculosisLigands from MycobacteriaLigands from YeastZymosan-TLR2/6(a beta-glucan)Ligands from VirusesCpG-DNA-TLR9dsRNA-

38、TLR3ssRNA-TLR7/8ssRNA/dsRNA-RLRsssDNA/dsDNA-cGASBoth DNA and RNA viruses vary greatly in both size and shapeTwo types of virusesSchematic drawings of viral genomes(A) Electron micrograph of an animal cell from which six copies of an enveloped virus (Semliki forest virus) are budding. (B) Schematic d

39、rawing of the envelope assembly and budding processes. The lipid bilayer that surrounds the viral capsid is derived directly from the plasma membrane of the host cell. In contrast, the proteins in this lipid bilayer (shown in green) are encoded by the viral genome.Acquisition of a viral envelopeFour

40、 virus uncoating strategies10 TLRs in Human, 11 in mouseNFBTLR signalingReceptorAdapterKinaseKinaseTranscription factorCytokinesIKK /TAK1TLRsMyD88IRAKTRAF6IRAK4MyD88TRAF6IRAKIRAK4IKKIKKIKKNFBTAK1Proinflammatory cytokines & chemokinesMyD88-dependentTLR2/1TLR2/6TIRAPMyD88 Pam3-CysMALP-2LTAZymosan TLR5

41、/7/8/9MyD88IKKIKKIKKNFBTRIFIRF3/7TBK1IKKiTRAF6TRAF3TAK1Type I-IFNProinflammatory cytokines & chemokinesTLR3MyD88-independentTRIF/LPS2TLR4 LPSTIRAPMyD88TRAF6IRAKIRAK4IKKIKKIKKNFBTAK1 LPSTRAMIRF3/7TBK1IKKiTRAF6TRAF3Type I-IFNLate activationProinflammatory cytokines & chemokinesMyD88-dependent and inde

42、pendentTRIFMyD88TRIF/LPS2TAK1TAB1TAB2TAK1MyD88PPTRAF6IRAKIRAK4IRAKTRAF6PPTAB1TAB2TAK1TRAF6IRAKPPUUNFBIKK /Complex IComplex IIComplex IIIThe complex regulation of TLR signalingNucleic acid-induced Innate ImmunityTLR3,7/8,9RLRHelicaseCARDDNA/RNAsAIM2dsDNAdsRNA5-pp-ssRNAMyd88/TRIFMAVS/VISASting/MITAERI

43、SNFBIRF3/7InflammasomedsDNAssDNATLR3: dsRNATLR7/8: ssRNATLR9: CpGDNAIntracellularAll viruses were detected by its genomic DNA/RNAsTakeuchi & Akira, 2010, CellTLR7/9ssRNA & CpG-DNATakeuchi & Akira, 2010, CellRLH dependent:Innate immunity to cytosolic Nucleic Acids from microbesNFBIRF3/7CARDRLRsHelica

44、seCARDCARDMAVSRNATBK1IKKiIKK/MitHow cytoplasmic DNA activates innate immunity?IRF3/7TBK1IKKiDNAIFI16DDX41cGASATP+GTPERSTINGNew pathwayOld pathwayThe most exciting findingin 2013: cGAMPs are ligands activating STINGDNAIFI16DDX41cGASATP+GTPcGAMPIRF3/7STINGCyclic di-nucleotides cGAMP: cyclic GMP-AMPJam

45、es Zhijian Chen(陳志堅)c-di-GMPc-di-AMPc-GAMPs(c-GMP-AMP)Cyclic di-nucleotides: New 2nd messengers, all activate innate immunityCyclic GMP-AMPs 22-cGAMP23-cGAMP32-cGAMP33-cGAMP 33-cGAMP 23-cGAMP32GMPAMPGMPAMPBackgroundDncVATP+GTP33-cGAMP InfectivityChemotaxis&Colonization12PDEs for c-di-GMP or c-di-AMP

46、 were identifiedWhat is the phosphodiesterase (PDE) that degrades cGAMP?STING + DncVIFN activation (fold)*Identification of a PDE candidate that inhibits DncV-induced IFN up-regulationHD-GYPHD-GYPHD-GYPHD-GYPHD-GYPHD-GYPHD-GYPHD-GYPHD-GYPAmong 9 HD-GYP proteins, 3 inhibit 33-cGAMPV. cholerae c-GAMP

47、phosphodiesterase, V-cGAP1/2/3123456789VCA0681VC2340VC1348VC1295VCA0931VC2497VCA0895ConVC1081VCA0210(226-458aa)(50-982aa)33-cGAMPVCA0681VC2340VC1348VC1295VCA0931VC2497VCA0895ControlVCA021033-cGAMPVC108133-cGAMPP1P2GAPDHp-IRF3DncVATP+GTPV-cGAP1V-cGAP15-pApGV-cGAP15-ApGV-cGAP1V-cGAP1 degrades 33- cGAMP to 5-ApG via the intermediate 5-pApGP1P2(PDEs)(5-nucleotidases)TLRsTLR3,7/8,9RLRHelicaseCARDDNA/RNANucleic acidsAIM2dsDNAdsRNAMyd88/TRIFMAVS