浙江大学医学免疫学经典课件免疫9、13-B细胞.ppt

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1、Lie Wang(汪洌）浙江省杭州市浙江大学紫金港校区医学院科研楼A810-814医学院免疫学研究所B lymphocytes and Humoral Immune ResponseThe discovery of B cell immunity1954-Bruce Glick,Ohio State UniversityStudies on the function of the bursa of Fabricius,a lymphoid organ in the cloacal region of the chickenBursa was later found to be the orga

2、n in which antibody producing cells developed antibody producing cells were thereafter called B cellsMammals do not have a bursa of FabriciusTransfer marked foetalliver cellsOrigin of B cells and organ of B cellmaturationNo MatureB cellsAfter birth,development continues in the bone marrowNormal bone

3、 marrowDefective bone marrowMature markedB cellsin peripheryB cell development starts in the foetal liverB cell development in the bone marrowBRegulates construction of an antigen receptorBone Marrow provides aMATURATION&DIFFERENTIATION MICROENVIRONMENTfor B cell developmentEnsures each cell has onl

4、y one specificityBChecks and disposes of self-reactive B cellsBExports useful cells to the peripheryBProvides a site for antibody productionBSecretedFactors-CYTOKINES2.Secretion of cytokines by stromal cellsBBone marrow stromal cells nurture developing B cellsTypes of cytokines and cell-cell contact

5、s needed at each stage of differentiation are differentStromal cell1.Specific cell-cell contacts between stromal cells and developing B cellsCell-cell contactBone marrow stromal cellMaturing B cellsBBStromal cellB lineage commitmentHSC(hematopoietic stem cell)MPP(淋巴系髓系多能前体细胞淋巴系髓系多能前体细胞)ELP(earliest

6、lymphocyte progenitor)ETP(early T-lineage progenitor)CLP(common lymphoid progenitor)HSCHSCHSCHSCDevelopment and migration of B cells(An overview)Stages of differentiation in the bone marrow aredefined by Ig gene rearrangementB CELL STAGEIgH GENECONFIGURATIONStem cellEarly pro-BLate pro-BLarge pre-BG

7、ermlineDH to JHVH to DHJHVHDHJHPre-B cellreceptorexpressedIg light chain gene has not yet rearrangedB cell receptorTransiently expressed when VHDHJH CHm is productively rearrangedIga&Igb signaltransductionmoleculesCHmHeavy chainVHDHJHLight chainVLJLCLVpreBl5Pre-B cell development is coupled with rea

8、rrangement of heavy-chain B cell development is coupled with rearrangement of heavy-chain B cell development is coupled with rearrangement of heavy-chain B cell development is coupled with rearrangement of heavy-chain B cell development is coupled with rearrangement of heavy-chain B cell development

9、 is coupled with rearrangement of heavy-chainSplicing of IgM and IgD RNACm1Cm2Cm3Cm4Cd1Cd2Cd3pA1VDJCm1Cm2Cm3Cm4Cd1Cd2Cd3VDJAAA RNA cleaved and polyadenylated at pA2VDJ CmIgM mRNAVDJ CdIgD mRNAC1C3CdCmVDJCm1Cm2Cm3Cm4Cd1Cd2Cd3DNApA1pA2VDJTwo types of mRNA can be made simultaneously in the cell by diff

10、erential usage of alternative polyadenylation sites and splicing of the RNARNA cleaved and polyadenylated at pA1AAA B cell development is coupled with rearrangement of heavy-chain6000 heavy chains combined with 320 light chains 1.9X106 Abs Ligation of the pre-B cell receptor1.Ensures only one specif

11、icty ofAb expressed per cellLargePre-BStromal cellUnconfirmed ligand of pre-B cell receptor2.Triggers entry into cell cycleALLELIC EXCLUSIONExpression of a gene on one chromosome prevents expression of the allele on the second chromosome1.Suppresses further H chain rearrangement2.Expands only the pr

12、e-Bcells with in frame VHDHJH joinsEvidence for allelic exclusionAllotypes can be identified by staining B cell surface Ig with antibodiesa/ab/ba/bYBbYBaYBbYYBabYBaANDALLOTYPE-polymorphism in the C region of Ig one allotype inherited from each parentSuppression of H chain rearrangement by pre-B cell

13、 receptor prevents expression of two specificities of antibody per cell(Refer back to Dreyer&Bennet hypothesis in Molecular Genetics of Immunoglobulins lecture topic)YYYYSuppression of H chain gene rearrangementensures only one specificity of Ab expressed per cell.Allelic exclusion prevents unwanted

14、 responsesBSelf antigenexpressed bye.g.brain cellsS.aureusYYYYYBS.aureusYYYYYYYAntiS.aureusAntibodiesYYYYYYAntibrainAbsOne Ag receptor per cellIF there were two Ag receptors per cellYYYYYYYAntiS.aureusAntibodiesPrevents induction of unwanted responses by pathogensAllelic exclusion is needed for effi

15、cient clonal selectionAll daughter cells must express the same Ig specificityotherwise the efficiency of the response would be compromisedSuppression of H chain gene rearrangement helps prevent the emergence ofnew daughter specificities during proliferation after clonal selectionS.typhiAntibodyS.typ

16、hiAllelic exclusion is needed to prevent holes in the repertoire Exclusion of anti-brain B cells i.e.self toleranceYYB BOne specificity of Agreceptor per cellS.aureusAnti-brain IgANDanti-S.aureus IgYYYB BIF there were two specificitiesof Ag receptor per cellAnti-brain IgBBDeletionAnergyORanti S.aure

17、us B cells will be excluded leaving a“hole in the repertoire”BUTYYYB B1.Suppresses further H chain rearrangementLigation of the pre-B cell receptor1.Ensures only one specificity ofAb expressed per cellLargePre-BStromal cellUnconfirmed ligand of pre-B cell receptor2.Triggers entry into cell cycle2.Ex

18、pands only the pre-Bcells with in frame VHDHJH joinsLargePre-BLargePre-BLargePre-BLargePre-BLargePre-BLargePre-BLargePre-BLargePre-BLargePre-BLargePre-BProliferationYImmatureB cellLight chain expressedIgM displayed on surfaceIgMLigation of the pre-B cell receptor triggers entry into the cell cycleLa

19、rgepre-BMany large pre-B cells with identical pre-B receptorsLarge pre-BIntracellular VDJCH chainVL-JL rearrangesProliferation stopsPre-receptor not displayedSmall pre-BBYYYYBSmall pre-B cellNo antigen receptor at cell surfaceUnable to sense Ag environment!May be self-reactive!Immature B cellCell su

20、rface Ig expressedAble to sense Ag environmentCan now be checked for self-reactivityAcquisition of antigen specificity creates a needto check for recognition of self antigens1.Physical removal from the repertoireDELETION2.Paralysis of functionANERGY3.Alteration of specificityRECEPTOR EDITINGB cell s

21、elf tolerance:clonal deletionImmature B cell recognisesMULTIVALENTself AgBClonal deletion byapoptosisY YBImmatureBBSmallpre-BSmall pre-B cellassembles IgYB cell self tolerance:anergyBYYYBAnergic B cellIgD normal IgM lowImmature B cell recognisessoluble self AgNo cross-linkingYYBImmatureBBSmallpre-BS

22、mall pre-B cellassembles IgIgMIgDIgDIgDReceptor editingA rearrangement encoding a self specific receptor can be replacedVCD JVVVYBB!Receptorrecognisesself antigen!BApoptosisor anergyYBBEdited receptor now recognisesa different antigen and can berechecked for specificityCD JVVVVArrest developmentAnd

23、reactivateRAG-1 and RAG-2YYYYYYMature B cellexported to theperipheryYYB cell self tolerance:export of self tolerant B cellsIgD and IgM normalIgMIgDIgDIgDIgDIgMIgMIgMImmature B cell doesnt recognise anyself AgYYBImmatureBBSmallpre-BSmall pre-B cellassembles IgBB-cell surface markers1.BCR complexBCR(m

24、Ig):VH,VL-Ag binding site mature B cells:mIgM and mIgD.Function:specifically recognizes antigen.Ig/Ig (CD79a/CD79b):heterodimer cytoplasmic domains contain ITAM.Function:transduce the signals that lead to B cell activation.Antigen receptor-mediated signal transduction in B cellsCD19/CD21/CD81complex

25、 CD21=CR2,C3dR,EB virus receptor CD19/CD21/CD81 interactions with complement associated with antigen play a role in antigen-induced B-cell activation.2.Co-receptorsThe role of the coreceptor in B cell activation(1)CD40 interacts with CD40L(Th cell)(2)CD80(B7.1),CD86(B7.2)Expressed on activated B cel

26、ls and other APCs(3)ICAM-1（CD54）、LFA-1（CD11/CD18)：mediate cell-cell interaction and co-stimulation3.Co-stimulatory molecules(1)CD20:function is unclear.It is suspected that it acts as a calcium channel in the cell membrane(2)CD22：Inhibitory receptor with ITIM motif(3)CD32(FcRII)：Inhibitory receptor(

27、4)Cytokine receptors(5)Complement Receptors(6)Toll-like receptors(7)MHC 4.Other receptors3.Subtype of B cells1.Conventional B cells(B-2 cells)2.B-1 cells(expression of CD5)CD5Two B cell lineagesBB cell precursorBMature B cellB2 B cellsPlasma cellYYYYYYYYYPCIgGBYYYYYYYYYYYYYYYYYYYYIgM-no other isotyp

28、esBDistinct B cellprecursor?B1 B cellsPrimitive B cells found in pleura and peritoneumB-2 cells:conventional B cells Recirculating follicular B cells:circulate between LN follicles and bloodmIg:IgM,IgDProduce IgG after antigenic stimulation in the presence of T helper cells B1 cells(CD5+):Many of th

29、e first B cells that appear during ontogeny express CD5,a marker originally found on T cells.(express mIgM,no mIgD).They respond well to TI-Ag and may also be involved in the Ag processing and presentation to T cells.Functions 1.produce anti-bacterial IgM the first line of defence against microorgan

30、isms;2.produce polyreactive Ab clearance of denatured self components;3.produce auto-Ab,thereby participating in the pathogenesis of some autoimmune diseases.Comparison of B-1 and B-2 B cell propertiesPropertyB-1 cellsB-2 cellsN regionsFewExtensiveV region repertoireRestrictedDiverseLocationPeritone

31、um/pleuraEverywhereRenewalSelf renewal in situBone marrowSpontaneous Ig productionHighLowIsotypesIgMIgM/G/A/D/ECarbohydrate specificityYesRarelyProtein specificityRarelyYesNeed T cell helpNoYesSomatic hypermutation of IgNoHighMemory developmentNoYesYes RarelyRarely YesNo YesCarbohydrate specificityP

32、rotein specificityNeed T cell helpSpecificity&requirement for T cell help suggests strikingly different typesof antigens are seen by B-1 and B-2 B cells4.Function of B cells1.Production of antibody Abs prevent microorganism from entry into cells and eliminate microorganisms by opsonization causing p

33、hagocytosis,complement activation and toxin neutralization.2.Ag presentation to T cells3.Immune regulation Secretion of cytokines(TNF,IFN,IL-12)M,DC,NK,B cell.Co-stimulation of T cellsT cell proliferation.YYImmune effector mechanisms against extracellular pathogens&toxinsNEUTRALISATIONYToxin release

34、blockedPreventstoxicityNEUTRALISING ANTIBODIESAdhesion tohost cells blockedPreventsinvasionBacteriumYToxinFc receptorbindingEffector mechanisms against extracellular pathogensOPSONISATIONOPSONISATIONPhagocytosisBacteria in extracellular spaceAb+Effector mechanisms against extracellular pathogensCOMP

35、LEMENT ActivationBacteria in plasmaAb&COMPLEMENT+PhagocytosisbindingComplement&Fc receptorLysisOpsonisationADCCB cell-mediated humoral immune responseHumoral immunity is mediated by antibodies and is the arm of the adaptive immune response that functions to neutralize and eliminate extracellular mic

36、robes and microbial toxins.It is more important than cellular immunity in defending against microbes with capsules rich in polysaccharides and lipids.TD-Ag：T cell-dependentTI-Ag：T cell-independentResponse to TD-Ag1)B cells recognize TD-Ag a.BCR directly recognizes B cell epitopes b.Ig/Ig transfer th

37、e first signalc.Signaling pathwaysd.Effect of coreceptors(CD21/CD19/CD81)Transduction of signals by the B cell receptorIgaIgbIntracytoplasmicsignalling domainsExtracellular antigenrecognition domainsThe cytoplasmic domains of the Iga and Igb contain Immunoreceptor Tyrosine-based Activation Motifs(IT

38、AMS)-2 tyrosine residues separated by 9-12 amino acids-YXXL/VX6-9YXXL/VBCR Signaling IBCR Signaling IIThe role of co-receptors(CD19/CD21/CD81)in B cell activation2)Role of Th cells in humoral immune response to TD-AgFor a protein Ag to stimulate Ab response,B cells and Th cells specific for that Ag

39、must come together in lymphoid organs and interact in a way that stimulates B cell proliferation and differentiation.a.Activation and migration of helper T cellsTh cells that have been activated to differentiate into effector cells interact with antigen-stimulated B cells at the edges of lymphoid fo

40、llicles in the peripheral lymphoid organs.The interactions of Th cells and B cells in lymphoid tissues.b.Presentation of Ags by B cells to Th cellsB cells that bind protein Ags by their BCR endocytose these Ags,process them in endosomal vesicles,display MHC II-peptides for recognition of Th cells.B

41、cells and Th cells recognize different epitopes of the same protein Ag.Ag presentation by B cells to Th cellsc.Mechanisms of Th cell-mediated activation of B cellsTh cells that recognize Ag presented by B cells activate B cells by expressing CD40L and by secreting cytokines(IL-2,IFN-,IL-4,IL-5,IL-6,

42、IL-13,etc.).Mechanisms of Th cell-mediated activation of B cellsKey components of T cell helpCD40L triggers CD40-synergizes with BCR signals to promote mitosis;cytokine(e.g.IL4)signals also contributeFasL triggers Fas-BCR signaling protects from apoptosisT cell derived cytokines influence differenti

43、ation,isotype switching:IL2,IL6 promote differentiationIL4 -IgG1,IgEIFN -IgG2a,IgG3TGFb and IL5 IgAMany other molecules involved in T-B interactions e.g.ICAM1/LFA1,ICOS/ICOSL,CD30/CD30L,CD27L/CD27,OX40L/OX40,.d.Th cells stimulate B cells to produce Abs of different heavy chain classes(isotypes)Heavy

44、 chain class switching is initiated by CD40L-mediated signals,and switching to different classes is stimulated by different cytokines.Ig heavy chain class(isotype)switching（蠕虫）Ig heavy chain class(isotype)switchingMechanisms of Ig heavy chain class switchinge.Affinity maturation in Ab responsesAffin

45、ity maturation is the process by which the affinity of Abs produced in response to a protein Ag increases with prolonged and repeated exposure to that Ag.The increase in affinity is due to point mutations in the V regions,and particularly in the Ag-binding HVR,of the Abs produced.Affinity maturation

46、 occurs in the germinal centers of lymphoid follicles.Affinity maturation in antibody responses somatic mutationControl of Affinity&Affinity MaturationFive B cell antigenreceptors-all specificfor ,but withdifferent affinitiesdue to somatichypermutationof Ig genes in the germinal centreBBBBBOnly this

47、 cell,that has a high affinity for antigen can express CD40.Only this cell can receive signal 2Only this cell is rescued from apoptosis i.e.clonally selectedThe cells with lower affinity receptors die of apoptosis by neglect Affinity maturation in antibody responses-Selection of high affinity B cell

48、sThe anatomy of humoral immune responsesA fraction of the activated B cells,which are often the progeny of class-switched high-affinity B cells,do not differentiate into Ab secretors but instead become memory B cells.Germinal CentersFunction:to generate B cells that produce antibodies with increased

49、 affinity for the inducing antigen=affinity maturationGerminal Center Reaction:Activated B cells give rise to Centroblasts（中心母（中心母细胞）胞）-localize in follicle,undergo rapid cell division and turn on machinery that causes somatic mutation in V-regionsCentroblasts give rise to Centrocytes（生（生发中心中心细胞）胞）-

50、migrate to the FDC-rich region of the Germinal Center-survival is dependent on interaction with FDC-bound Ag and presentation of Ag to T cells-centrocytes that successfully compete to bind antigen(e.g.by having higher affinity BCR)and to receive T cell help are selected and may differentiate into lo