1、【医学英文课件】Cytokines and Lymphocyte Subsets No Antigen DNP-GPA DNP-BCG DNP-BADNP-GPA T cell mediaDNP-BCG T cell media System for cell communication in the absence of cell:cell contact J David/B Bloom,1966Cytokines:Discovery of MIFSolubleCell ContactCytokines inform cellsCytokine functionsCell-cell comm
2、unicationResponse amplificationResponse regulationCytokine Function-ExamplesCell Growth/IL-3,GM-CSF,G-CSF,M-CSF,IL-34Differentiation IL-2,IL-5,IL-7,IL-15,IL-21Cell DeathTNF,FasLInflammationIL-1,IL-6,TNF,IL-8,IL-12 IL-17,IFN-g g,IL-18Immune RegulationIL-4,IL-10,TGFb bCytokine biosynthesis-general fea
3、tures1.Absent from unstimulated cells.2.Rapid transcriptional activation with stimulation.3.Can have post-translational control;TNF,TGF-b b.4.Rapid secretion.5.Short-lived transcripts and proteins.6.Transcription ceases when stimulus abates.Cytokine headaches1.How many?2.Who named these(TNF,RANTES,e
4、tc.)?3.How do you get to be an IL?4.Redundancy and Pleiotropy-do we care about any individual cytokine?Do we really know what some of these do?Regulated upon activation,normal T cell expressed and presumably secretedCytokine headachesScience 2016 Official Interleukin Names:HUGO(Human Genome Organiza
5、tion)and IUIS(International Union of Immunological Sciences)HUGO=Genomics basis IUIS=Functional basishttp:/www.gene.ucl.ac.uk/nomenclature/genefamily/interleukins.htmlhttp:/www.genenames.org/genefamily/il.phpCytokine solutions I.Order in structurally-related Families-reflects origin through gene dup
6、lication and inter-chromosomal spreadAdvantages:Common Receptor FamiliesCommon Signaling AdaptersFunctional LinkageGenomic LinkageCytokine solutions II.Order in functionally-related Families:Inflammatory/Early:IL-1,IL-6,TNF,IL-12,IL-17Growth/Differentiation:CSF1-3,Epo,IL-7,kit-LMigratory:ChemokinesI
7、mmuno-activating:IFNs,IL-4 Immuno-de-activating:TGFb,IL-10,soluble receptorsAdvantages:Biologically grouped,easier to remember 4a Helix:IL-2 and gc superfamily(IL-2,4,7,9,13,15,21,TSLP)IL-6/gp130 superfamily (IL-6,IL-11,G-CSF,leptin,LIF,OSM,CNTF,CT-1,IL-31)IL-12/gp130-related superfamily(IL-12,IL-23
8、,IL-27,IL-35)IFN/IL-10 superfamily:(IFNa/b,IFNg,IFNl,IL-10,19,20,22,24,26)Not called interleukins:CSFs,LIF,Flt-3L,EPO.Non-interleukins:GH1,PRL,LPT,.IL-1 family:IL-1F1(IL-1a),IL-1F2(IL-1b).IL-1F4(IL-18)IL-1F5(IL-36Ra)IL-1F6/8/9(IL-36a/b/g).IL-1F7(IL-37)IL1F11(IL-33)IL-17 family:IL-17AIL-17F,IL-25TNF
9、family:TNFSF1(LTa).TNFSF18(AITR-L)Chemokine family:CXC,CC.,C(lymphotactin 1/2),CX3C(fractalkine)Major Cytokine Families ShortLongAAD DBCCNNCAutocrineAutocrineParacrineParacrineIL-2,4,7,9,13,15,21EndocrineIL-3,5,GM-CSF IL-6 familyImmune System IL-12 p40 family Hormones,EPO,TPO BType 1 cytokines:4a a-
10、helix bundleFBDCEECDBFA A4a a-Helix FaceIFN a a/b bIFN-g g/IFN-l l 1-3IL-10 superfamilyType 2 cytokine family-6 helix structureBox 1 (JAKs)Box 2(STATs)Conserved cysteinesWSXWS motifType I Cytokine Receptors Type II Cytokine Receptors4a a helix cytokines bind to cytokine receptor superfamily members
11、R1R1R2R2Paradigm for cytokine/cytokine receptor interactions R1R1R2R2Paradigm for cytokine/cytokine receptor interactions R1R1R2R2Paradigm for cytokine/cytokine receptor interactions JAKJAKUsually constitutively associated with JAK-box motif(JAK3 can be induced)Four JAKs:JAK1,JAK2,JAK3,Tyk2Type 1 an
12、d type II cytokine receptors signal via JAKs and STATs JAKJAKReceptor OligomerizationPPYYTyrosine phosphorylation of cytokine signaling chainType 1 and type II cytokine receptors signal via JAKs and STATs STAT JAKJAKReceptor OligomerizationPYPYSTATPYRecruitment and phosphorylation of STAT(dimers)on
13、tyrosineSTAT:signal transducers and activators of transcriptionLatent cytosolic transcription factorsSeven STATs:STAT1,2,3,4,5a,5b,6Type 1 and type II cytokine receptors signal via JAKs and STATs JAKJAKReceptor OligomerizationPYPYSTATPYPYSTATSTAT nuclear translocation,gene transcriptionType 1 and ty
14、pe II cytokine receptors signal via JAKs and STATs Common paradigms underpinning redundancyUnique recognition receptor chainCommon signaling receptor chainCombinatorial use of eachCombinatorial use underpins cytokine redundancyType 1 cytokine SF Common chain a chain Adaptergc 2,4,7,9,15,21 JAK-3bc 3
15、,5,GM-CSF JAK-2gp 130 IL-6 familyJAK-1,JAK-2gp-130-like IL-12 p40 familyJAK-2,Tyk-2Combinatorial use underpins cytokine redundancyType 2 cytokine SF a chains Adapters IFNAR1,2 Tyk-2,JAK-1IFNGR 1,2JAK-1,JAK-2IL-10RaJAK-1IL-10RbTyk-2 Hormones(dimeric a chains)JAK-2Combinatorial but engagement sequence
16、 mattersLePorte,SL et al.Cell 132:259-272,2008.IFN-a a/b b IFN-g g IL-28/29 IL-10 IL-19/IL-20/IL-24 IL-22 IL-20/IL-24 IL-26 (IFN-l l1-3)IFNAR1/IFNGR1/IFNGR1/IL-10R1/IL-20R1/IL-22R/IL-22BP IL22R/IL-20R1/IFNAR2 IFNGR2 IL-10R2 IL-10R2 IL-20R2 IL-10R2 IL-20R2 IL-10R2 Jak1/Tyk2 Jak1/Jak2 Jak1/Tyk2 Jak1?J
17、ak1/Tyk2 Jak1?Tyk2?STAT1-3 STAT1 STAT1/3 STAT3/5 STAT3/5?STAT3/5Chromosome 9 12 19 1 1/1/1 12 1/1 12 21 21 6 21 6 21 11 21 6 3 1 21 6 1 3 21IFN/IL-10 superfamily of cytokines and cytokine receptorsCombinatorial but receptors matterOuyang W et al.Regulation and functions of the IL-10 family of cytoki
18、nes in inflammation and disease.Annu Rev Immunol 29:71-109,2011.StructureReceptor Family AdaptersTIR DomainIg DomainsCRDsTRAFsDDSer-Thr Kinaseb b-Trefoil(IL-1F)Jellyroll(TNF)Cysteine Knot(TGF-b b)MyD88 MAL,TRIF,TRAM,SARAM;IRAKs;IRFs DD,TRAFs 1-6 SMADsStructural Paradigm JAKJAKJAKJAKStatStatStatStatS
19、tatSOCSSOCS proteins:negative regulators of cytokine SFRsSOCS proteins:negative regulators of cytokine SFRsLinossi EM,SE Nicholson.2015.Immunological Reviews 266:123-133.Adhesins,ActivationMigrationNeutrophils then Monocytes,Acute Phase Response,FeverDC Activation,Antiviral stateLN,SpleenIL-1,TNFChe
20、mokinesIL-6,IL-17IFN-b b InitiatorsIFN-g g,IL-4 EffectorsIL-10TGFb bRegulators Take Home Message:Cytokines Mediate ImmunityA Working ModelTherapeutic Potential of CytokinesHarnessing cytokines as therapeuticsAnti-TNF,IL-1,IL-6,IL-12/23,IL-5,IL-13,IL-17Hematopoietic growth factors for stem cell mobil
21、izationJAK inhibitors(JAKinibs)Insights from cytokinopathiesGenetic loss(IFNg gR,IL-12,etc),autoantibodies(GM-CSF,IFNg g,IL-17,etc),interferonopathies(deficiencies of Trex1,MDA5,STING,ISG15,etc),NLRP3(hereditary periodic fevers,etc)Th1Th2IFN-g gIL-2LymphotoxinIL-4IL-5IL-13IL-10IL-9Th subsets:stable
22、mouse clones(Mosmann,Coffman-1986)Critical early concepts driven by Th1/Th2 paradigmCytokines mediate immune effector functions of helper T cellsHelper T cells produce patterns of cytokines that mediate coordinate responses by effector cellsDistinct pathogens elicit distinct Th cytokine patternsEffe
23、ctor patterns become stabilized in recall responsesTh1/Th2-definitionsOriginalStable CD4+clonesGeneral UsageAny response polarized towards IFN-g or IL-4Operational ExtensionType 1 immunityHost response coordinated by Th1 cells and its downstream targets,mainly activated phagocytes,armed with complem
24、ent-fixing Ig(IgG2a in mouse),and CTLType 2 immunityHost response coordinated by Th2 cells and its downstream targets,mainly activated eosinophils and mast cells,armed with non-complement-fixing Ig(IgE,IgG1)Paradigms ExtendedCD8(Tc1/Tc2),NK,B cells,DC,etc.TSLPIL-33IL-25IL-13Th1Th2Systemic ImmunityPh
25、agocyte activationOpsonizing AbCTL,NK cellsBarrier ImmunityMast cells,basophilsEosinophilsIgE,IgAAllergy&asthmaIFNg gLTIL-4IL-5IL-9IL-13NaveCD4+T cellType 2Type 1IL-27IL-18IL-12IL-4 DCAntigen EliminationAntigen Sequestration DCEoBasoMast cellNKIFNg gThe Extended Th1/Th2 paradigmTh subsets:an epigene
26、tic differentiation modelCytokine loci are methylated and associated chromatin is hypo-acetylatedPolarizing conditions associated with chromatin acetylation,new DNase I hypersensitivity sites(1-2 days)and demethylation of target genes(4-7 days)Chromatin and DNA changes are epigenetically stabilized
27、in polarized clones(irreversible)Current Paradigm:Master Regulators Modulate Chromatin and Mediate Changes in DNA Methylation at Cytokine Genes by Targeting Transcriptional Machinery Bird et al.,Immunity 9:229-37,1998;Agarwal and Rao,Immunity 9:765-75,1998Conserved Noncoding Sequences in the type 2
28、cytokine cluster on chromosome 11(5q)Bold=Single-copy*=conserved in human,mouse,rat,rabbit,dog,cow,pig,chicken,fuguLoots et al,Science 2000IL-4 IL-13IL-5IL-3100 kb 460 kb8 kb4 kbCNS-1(401 bp)GM-CSFIL-4IL-13Conserved non-coding sequence CNS-1:regulatory element for IL-4 and IL-13conservation:IL-4/13
29、50%CNS-1 89%Loots et al,Science 2000CNS-1-/-mice have impaired Th2 responsesMohrs et al,Nat Immunol,2001 IL-13 RAD50IL-43 Enhancer Silencer Intronic Enhancer Promoter Intergenic Enhancer CD8 ROG site LCRTCR specificity Th2 Th2 specificity,weak enhancer TCR Strong enhancer CD8 Silencing PositionWeak
30、enhancer specificity Effect Notch target Lee et al.,Immunity 14:447-59,2001;Lee et al.,Immunity 19:145-53,2003;Omori et al.,Immunity 19:281-94,2003;Ansel et al.,Nat Immunol 5:1251,2004.Mini-locus of all elements enhanced by co-expression of GATA-3 CNS-1Regulatory elements in the IL-4/13 locusKM Anse
31、l et al,An epigenetic view of helper T cell differentiation.Nature Immunol 4:616-623,2003.Looping-out model for coordinate gene expression at the type 2 cytokine locusTissue-specific expression regulated by incorporation of LCR and induction of lineage-specific factorsLee GR et al:T helper cell diff
32、erentiation:regulation by cis elements and epigenetics.Immunity 24:369-79,2006.GATA-3T-betGATA-family zinc finger transcription factorDiscovered in Flavell lab using differential displayNecessary and sufficient for Th2 differentiationT-box family transcription factor,Tbx21Discovered in Glimcher lab
33、using yeast 1-hybridNecessary and sufficient for Th1 differentiationMaster regulators of Th differentiationInitiation of Transcription Stabilization HardwiredFrom Murphy and Reiner,The lineage decisions of helper T cells,Nature Rev Immunol 2002,2:933.Th2nave CD4+Th1IL-4g-satellitePositioning near ar
34、eas of condensed,nontranslated,chromatinTerminal differentiation in Th subsets is associated with repositioning of cytokine genescollagenmyelinarthritisEAEIL-12Rb b1 koIL-12/23 p40 koIL-12:p35/p40:IL-12Rb b1/IL-12Rb b2IL-23:p19/p40:IL-12Rb b1/IL-23RCD4 T cell-dependentAdditional CD4 subsetsIL-21IL-2
35、3Th1Th17Systemic ImmunityPhagocyte activationOpsonizing AbCTL,NK cellsInflammatory ImmunityNeutrophilsMonocytesIFNg gLTIL-17A,-17FIL-22,-26IL-21NaveCD4T cellType 17Type 1IL-12 DCAntigen EliminationAcute Inflammation DCIL-6TGFb bIFNg g-Th17 cells mediate neutrophil inflammationCiofani MDR Litmman.201
36、2.A validated regulatory network for Th17 cell specification.Cell 151:289-303.T cells differentiate using positive and negative regulatory modules(i.e.;there is no master regulator)Transcriptional networks direct epigenetic fate Ciofani MDR Litmman.2012.A validated regulatory network for Th17 cell s
37、pecification.Cell 151:289-303.Epigenetic fate directs phenotype position,function,lifespanCiofani MDR Litmman.2012.A validated regulatory network for Th17 cell specification.Cell 151:289-303.Transcriptional modules as paradigms for parsing positioning and effector functionJosefowicz S,L-F Lu,AY Rude
38、nsky.2012.Regulatory T cells:mechanisms of differentiation and function.Annu Rev Immunol 30:531-64Peripheral tissuesLymphoid tissuesNave CD4 T cellTFH Lots and lots of papersCritical for every successful human vaccineCD4 T cell help orchestrates immunityIL-4 PrimingIL-4 SecretionIL-13 SecretionTFHBE
39、DTAReinhardt et al.,Nat Immunol 10:385,2009:Lymph node IL-4-secreting T cells are TFH cellsIL-4 Mediastinal LN LungIL-13 Stat6 IL-4/13 Lee Reinhardt,Hong-Erh Liang Intracellular GATA-3 Bcl6 GATA-3loLN Cytokine production by TFH cells and Th2 cellsIL-13 IL-5 GATA-3hiPeriphery Blimp Tissue IL-13-secre
40、ting cells are Th2 effector cellsLung CD4 cells YFP+yfp-IL-5IL-13XROSA26 locus Hong-Erh Liang,Lee ReinhardtNimmerjahn and Ravetch,Immunity 24:19-24,2006 IL-4 IFNg gTGFb bTh2Th1Th17FcRs bind IgG heavy chains with different affinitiesTh subsets coordinate distinct populations of innate cells that shar
41、e effector functionsTh2 cells:Basophils,eosinophils,mast cells,alternatively activated macrophages,(IgE,IgG1)Th1 cells:NK cells,CD8 T cells,activated macrophages,(IgG2a)Th17 cells:PMN,monocytes,(IgG2b)Implies role for Th cells in attraction,organization,sustaining survival of myeloid cells.Innate Ly
42、mphoid Cells,2016Lineage-negative,Id2-dependent cells that arise from a common lymphoid precursor.Comprise cells that mediate lymph node organogenesis during fetal development and become tissue-resident effector cells expressing cytokine programs driven by transcriptional modules associated with typ
43、ical CD4 helper subsets(Th1,Th2,Th17,etc).Roles in homeostasis(establishing commensals,responding to dietary signals,responding to circadian cues,etc).Roles in inflammation and disease(facilitate programs mediated by established Th subsets).Group 1:T-bet+IFNGroup 2:GATA3hiGroup 3:RORt+IL-5 IL-13IL-1
44、7A IL-22Ex-RORtIEL ILC1LTiNK-22ILC1cNKIFNNatural helper cellsNuocytesIh2 ILC1 ILC2 ILC3 IL-12,IL-15,IL-18,etc.IL-33,TSLP,IL-25,etc.IL-1,IL-6,IL-23,Ahr ligands,etc.,Th1VTh2VTh17Th22VVEx-Th17VInnate TCR+lymphoid cells Berg LJ,Signalling through TEC kinases regulates conventional versus innate CD8+T ce
45、ll development.Nat Rev Immunol 7:479,2007.McDonald BD et al.,Nature Immunol 14:1110,2013 Cell 164:1198,2016Cell 150:1235-48,2012Kastenmuller WRN Germain,Cell 150:1235-48,2012.Localization of invariant effector lymphocytes in draining LNsonSherlock JPDJ Cua,Nature Med 18:1069-76,2012.Localization of
46、invariant effector lymphocytes at sites of tissue stressonIL-17-producing gdgd T cells in tendon enthesesModified from Eberl&Sawa,Trends Immunol 31:50-5,2010714210neonatal flora/metabolitesadult flora/metabolitesdays post-birthRORg g/IL7R/g gcLTa a/LTb bRRANKL(SI)AHRMicrobiota/MyD88/NOD1IL-22CCR6/CC
47、R7/CXCL13a a4b b7/MAdCAM1Inductionof CPsActivationOf ILC cytokinesInductionILFs?weanCPiILFmILFILC3Modified from Eberl&Sawa,Trends Immunol 31:50-5,2010714210neonatal flora/metabolitesadult flora/metabolitesdays post-birthRORg g/IL7R/g gcLTa a/LTb bRRANKL(SI)AHRMicrobiota/MyD88/NOD1IL-22CCR6/CCR7/CXCL
48、13a a4b b7/MAdCAM1Inductionof CPsActivationOf ILC cytokinesInductionILFs?weanCPiILFmILFILC3Catch22 colonHomeostasisTGF-b b,retinoic acid,etcTreg ILCIL-10,IL-35PerturbationIL-6/IL-1 plus TGF-b b,retinoic acid,etc (+/-adaptive immune response)TregILCIL-17,etcRepairIL-6(TGF-b b exhausted from environme
49、nt)TregILCIL-22,IL-10,etcHomeostasisTGF-b b,retinoic acid,etc ILCTregIL-10,IL-35Resident memory adaptive effectors displace innate effectors at peripheral sites of injuryZaid et al,PNAS 111:5307,2014Immune SystemTissue HomeostasisCommensals and PathogensLatent organismsStem CellsClarity from darknessMembers of the Locksley Lab at UCSF
