固有免疫研究進(jìn)展課件

固有免疫研究進(jìn)展固有免疫研究進(jìn)展Contents1、Innateimmunesystem2、Innateimmunerecognition3、Innateimmuneresponse4、ProgressofinnateimmunityContents1、Innateimmunesystem固有免疫研究進(jìn)展課件固有免疫研究進(jìn)展課件InnateImmunitySystem一、固有免疫系統(tǒng)InnateImmunitySystem宿主固有免疫系統(tǒng)固有免疫系統(tǒng)固有屏障結(jié)構(gòu)皮膚與粘膜血腦屏障胎盤(pán)屏障固有免疫分子補(bǔ)體溶菌酶防御素固有免疫細(xì)胞PMNDCsMf宿主固有免疫系統(tǒng)固有免疫系統(tǒng)固有屏障結(jié)構(gòu)皮膚與粘膜血腦屏障胎（一）固有屏障結(jié)構(gòu)（一）固有屏障結(jié)構(gòu)Figure2-4Intactskin/MucusmembraneMucosalimmunesystem(MIS)Secretionofanti-microbialfactorsNormalfloraBlood-brainandplacentabarriersAnatomicandPhysiologicBarriersFigure2-4Intactskin/MucusmeBarriersofinnateimmunitysystemSkinandmucosalbarrierPhysical:skin&mucosa,epidermis-dermisconjunction,mucosalsecretion,lavagingoffluidsandciliaryactionChemical:acidicsecretion,lysozymeMicroorganism:commensalbacteriaBarriersofinnateimmunitysy固有免疫研究進(jìn)展課件LocalbarrierstructureBlood-thymus-barrierBlood-gas-barrierBlood-urine-barrierBlood-testis-barrierBlood-epididymis-barrierCerebralbarrier:BBB,etc.Blood-fetus-barrierMucosa-bicarbonatebarrierLocalbarrierstructureBlood-t（二）固有免疫細(xì)胞（二）固有免疫細(xì)胞ComponentsoftheImmuneSystemComponentsoftheImmuneSyste固有免疫細(xì)胞固有免疫細(xì)胞固有免疫研究進(jìn)展課件固有免疫研究進(jìn)展課件PhagocytosisTypeofphagocytes:Smallphagocytes(neutrophils)largephagocytes(macrophage)

DendriticCellsChemotaxisContactPhagocytosis

IntracellularkillingPhagocyticeffectsenhancedbyantibodyorcomplementPhagocytosisTypeofphagocytes吞噬和殺菌過(guò)程： 趨化

黏附

吞入

殺滅與消化(CD14)吞噬和殺菌過(guò)程：(CD14)Real-timemovieofphagocytosisReal-timemovieofphagocytosi（三）固有免疫分子（三）固有免疫分子固有免疫分子固有免疫分子固有免疫分子Complment:alternative&MBLpathwayLysozyme:tissueandbodyfluid,secre-tion,lysosomeinphagocytesAntimicrobialpeptides:defensinCytokines:interferon,IL-1,IL-6,IL-12,TNF-α,chemotaxinAcutephaseprotein:CRP,MBP,SAP,α1-acidglycoprotein,FN,etc.固有免疫分子Complment:alternative&LysozymeKillingG+bacteria,splitstheexposedpeptidoglycanwallCategory:BacteriallysozymeActonβ-1,4glycosidicbondActonthepeptidetailandamideFungallysozyme:

Saccharomycetic,AspergillarLysozymeKillingG+bacteria,sCytokinesInterferon:virallyinfectedcellscansynthesizetypeⅠIFN.ReducemRNAtranslation,degrademRNA,inhibitviralreplicationInterleukin:IL-1,IL-6,IL-12ChemotacticfactorsTNF:TNF-αCytokinesInterferon:virallyiInnateRecognition二、固有免疫識(shí)別InnateRecognitionFatherofInnateImmunity：Prof.JanewayCConcept:PRR:PathogenrecognizingreceptorsPAMPs:PathogenassociatedmolecularpatternsFatherofInnateImmunity：Prof（一）識(shí)別什么？PAMPs（一）識(shí)別什么？PAMPsPAMPandPRR1、PAMP:PathogenAssociatedMolecularPatternsLippolysaccharide(LPS),dsRNA,unmethylatedCpG,mannose,glucan,Lipoteichoicacid(LTA),Pepidoglycan(PGN)2、PRR:receptorsonprofessionalAPCandMφwhichenablethemtorecognizePAMP.mannosereceptorToll-likereceptorscavengerreceptorphosphatidylserinereceptorPAMPandPRR1、PAMP:PathogenA29PathogenAssociatedMolecularPatternPAMPareconserved,sharedbyalargegroupofinfectiousagents,andclearlydistinguishablefromselfpatterns.TypicalPAMP: lipopolysaccharide(LPS), mannose,glucan,dsRNA, lipoteichoicacid(LTA), peptidoglycan(PGN) unmethylatedCpGDNAetc29PathogenAssociatedMoleculaPathogen-AssociatedMolecularPatterns(PAMPs)Pathogen-AssociatedMolecular黏附素（adhesin)黏附素（adhesin)fimbrialadhesins(E.

coli)non-fimbraladhesin（Bartonella）黏附素fimbrialadhesins(E.coli)no固有免疫研究進(jìn)展課件LigandSpecificitiesofTLRsLigandSpecificitiesofTLRs（二）用什么去識(shí)別？PRR（二）用什么去識(shí)別？PRR3636Pattern-RecognitionReceptor，PRRExpressedmainlyonDCandMfwhichenablethemtorecognizePAMP.ThreetypesofPRR:secretedPRR:MBL,C-reactiveproteinendocyticPRR:

scavengerreceptor,mannosereceptorsignalingPRR:

Toll-likereceptor, NOD-likereceptor, RIG-likereceptor3636Pattern-RecognitionRecept3737一）TLR的發(fā)現(xiàn)Drosophilaembryogenesis.(1980s)AntifungalresponseinDrosophilaadults. (Hoffmannetal.,Cell,1996)AhumanhomologueofToll(Janewayetal.,Nature,1997)3737一）TLR的發(fā)現(xiàn)Drosophilaembryog3838The2011NobelPrize

JulesA.HoffmannBruceBeutlerTheyreceivedone-halfofthe2011NobelPrizeinPhysiologyorMedicine,for"theirdiscoveriesconcerningtheactivationofinnateimmunity"3838The2011NobelPrizeJulToll-likeReceptors(TLRs)Afamilyofproteins,whicharestructurallyrelatedtoDrosophilaTollandIL-1receptor.TLR1-10inhuman,TLR1-13inmouse.Recognizespecificpathogenassociatedmolecularpatterns(PAMPs)suchasendotoxin(LPS).Regulatetheactivationofbothinnateandadaptiveimmunity.Toll-likeReceptors(TLRs)Afa固有免疫研究進(jìn)展課件TLRsignaling

AkiraS.etal.(2005)Int.Immunol.17:1-14

TLRsignalingAkiraS.etal.固有免疫研究進(jìn)展課件4343TLRsininnateandadaptiveimmunityTLRsininnateandadaptiveimTLRscontroladaptiveimmunityAkiraS.etal.

(2001)NatureImmunology

2:675-680

TLRscontroladaptiveimmunity固有免疫研究進(jìn)展課件固有免疫研究進(jìn)展課件參與固有免疫識(shí)別的受體模式識(shí)別受體（PRR)參與固有免疫識(shí)別的受體抗感染免疫與固有免疫識(shí)別抗感染免疫與固有免疫識(shí)別NOD-likereceptors,NLRRecognizecytoplasmicPAMPsand/orendogenousdangersignals,inducingimmuneresponses.Ligands:MDP

andmeso-DAPThreedomains:C-terminalLRRssenseligandscentrallylocatedNODdomainN-terminaleffectorregioncomprisingaprotein-proteininteractiondomainsuchastheCARD,PyrinorBIRdomain.NOD-likereceptors,NLRRecogniNLR

subfamiliesSeveralsubfamilies(onthebasisoftheireffectordomains)CARDsubfamily:NODs,IPAF,CIITA,PYDsubfamily:NALPsBIRsubfamily:NAIPsNLRsubfamiliesSeveralsubfami52NLRDomains52NLRDomains53NLR

families53NLRfamiliesFunctionsofNLRTheymaythenformoligomersthatactivateinflammatorycaspase1causingcleavageandactivationofimportantinflammatorycytokines（IL-1β）NLRsmayalsoactivatetheNF-kB

signalingpathwaytoinduceproductionofinflammatorymolecules.FunctionsofNLRTheymaythen5555NLR

signaling

5555NLRsignalingNLRandinflammasomeinflammasome,amultiproteincomplexthatcontains,inadditiontoNLR,theadaptormoleculeASCandtheproteasecaspase-1.

inflammasomeisresponsiblefortheproteolyticprocessingoftheimmatureformofthecytokinesbelongingtothe

IL-1family

(IL-1b,IL-18,andIL-33),56NLRandinflammasomeinflammasIL-1β/IL-18inflammosome57IL-1β/IL-18inflammosome57NALP3andIPAFinflammasomeFabioMartinon，Annu.Rev.Immunol.2009.27:229NALP3andIPAFinflammasomeFabRIG-Ilikereceptors，RLRDetectintracellulardsRNAofviralorigin:RIG-I(retinoicacidinducedgene)MDA-5(melanomadifferentiationassociatedgene)LGP2(laboratoryofgeneticsandphysiology)ComprisesN-terminalcaspaseactivationandrecruitmentdomains(CARDs),aDECHhelicase,andaC-terminaldomain(CTD).RIG-Ilikereceptors，RLRDetect60RLRDomainsRedbox,CARD1;bluebox,CARD2;greenbox,Helicasedomain;purplebox,DEAD/DEAHdomain60RLRDomainsRedbox,CARD1;61SignalingofRLR61SignalingofRLRe.g.,HCVproteinNS3/4AeliminatesantiviralsignalingbycleavageofMAVSandTRIF.

e.g.,HCVproteinNS3/4Aelimin63SummaryofPatternRecognitionTheJournalofClinicalInvestigation,118(2),2008?63SummaryofPatternRecognitiInnateImmuneResponse固有免疫應(yīng)答InnateImmuneResponse固有免疫研究進(jìn)展課件固有免疫研究進(jìn)展課件固有免疫研究進(jìn)展課件固有免疫研究進(jìn)展課件6969固有免疫研究進(jìn)展課件固有免疫研究進(jìn)展課件視頻(ImmuneResponse)視頻固有免疫研究進(jìn)展課件固有免疫研究進(jìn)展課件固有免疫研究進(jìn)展課件固有免疫研究進(jìn)展課件IntracellularkillingmechanismsO2-dependent:ROI,RNI(reactivenitrogenintermediate),SOD(superoxidasedismutase)O2–independent:MPO(myeloperoxidase,)Enzymes(lysozyme)

DefensinsIntracellularkillingmechanisProgressofInnateImmunity固有免疫研究進(jìn)展ProgressofInnateImmunity抗細(xì)菌感染免疫的固有免疫識(shí)別與免疫應(yīng)答抗細(xì)菌感染免疫的抗病毒免疫的固有免疫識(shí)別與免疫應(yīng)答抗病毒免疫的P.aeruginosakeratitisNormalcorneaBacterialKeratitis:ClinicalPictureP.aeruginosakeratitisNormal研究依據(jù)WHO（2006）:在影響身心健康的疾病中，眼科疾病僅次于腫瘤與心血管疾病角膜病－－－主要的致盲性眼病 中國(guó)角膜盲患者350～500萬(wàn)

絕大部分與角膜潰瘍相關(guān)廣東省是角膜潰瘍的高發(fā)區(qū)研究依據(jù)WHO（2006）:在影響身心健康的疾病中，眼科疾病立項(xiàng)依據(jù)角膜感染 潰瘍穿孔

致盲感染性角膜潰瘍： 青壯年勞動(dòng)人口、綠膿桿菌病理本質(zhì)：免疫病理?yè)p害 潰瘍壞死臨床治療：抗感染

耐藥性、過(guò)度免疫反應(yīng) 潰瘍穿孔 致盲立項(xiàng)依據(jù)角膜感染 潰瘍穿孔 致盲免疫病理?yè)p害機(jī)制激活機(jī)體的抗感染免疫

清除病原體病原體感染宿主免疫激活PRR受體

過(guò)度炎癥反應(yīng) 免疫病理?yè)p害

LPS、DNA、鞭毛素

負(fù)調(diào)控免疫病理?yè)p害機(jī)制激活機(jī)體的抗感染免疫病原體感染宿主免疫

熱 紅

腫 痛功能喪失

炎癥熱 紅 腫 痛功能喪失炎癥BacteriaPseudomonasaeruginosaATCCstrain196601x106CFU/mlappliedtoscarifiedcorneaBacteriaPseudomonasaeruginosaAnimalmodelofP.aeruginosainfectionC57BL/6(B6)mice Susceptible(cornealperforationwithin5-7daysp.i.)BALB/cmice Resistant(restorecornealclaritywithin1-2wkp.i.)

AnimalmodelofP.aeruginosaMicroarrayanalysisWhatistheoverallgeneexpressionprofileinB6vs.BALB/cmiceafterP.aeruginosacornealinfection?

B6andBALB/cmicewereinfectedwithP.aeruginosa

Uninfected(N)corneastakenandat1dayp.i.TotalRNAextractedforlabelingRNAhybridizedwithamurinearrayMG-U74(Affymetrix)ArraychipsscannedforrawdataDataanalyzedusingbioinformaticssoftwarepackageHuangX.andHazlettLD.

(2003)IOVS44:3409-3416Microarrayanalysis B6andBALType-1(Red)vs.Type-2(Green)B61dpiCCEEDDFFB6NormalB6NormalBALB/cNormalBALB/cNormalB61dpiB61dpiBALB/c1dpiBALB/c1dpiBBBALB/cNormalBALB/c1dpiAAB6NormalBlue=DetectableRed=type-1genesGreen=type-2genesHuangX.andHazlettLD.

(2003)IOVS44:3409-3416Type-1(Red)vs.Type-2(GreenGeneexpressiondifferencesconfirmedbyPCR

HuangX.andHazlettLD.(2003)IOVS44:3409-3416GeneexpressiondifferencescoRNAexpressionofcellulargenesup-(ordown-)regulated4-foldorhigherinbothmousestrains

GeneName AccessionNo.TLRs/Signaltransduction:

1.CD14 X133332.IL-1R L328383.TLR-6 AB0208084.IKKi AB0165895.NIK U889846.Fos-likeantigen1 AF0171287.Fasantigen M836498.TNFReceptor1 X871289.TRAF-1 L35302HuangX.andHazlettLD.

(2003)IOVS44:3409-3416RNAexpressionofcellulargenTLR4inbacterialkeratitisP.aeruginosaLPS,aspecificligandofTLR4.

AkiraS.etal.(2005)Int.Immunol.17:1-14

TLR4inbacterialkeratitisP.DayspostinfectionN13570306090120150RelativemRNAofTLR4TLR4mRNAexpressioninBALB/cmiceHuangX.etal.(2006)IOVS47:4910-4916DayspostinfectionN1357030012345BALB/cTLR4lps-d1d3d5d

DayspostinfectionClinicalscoreDiseaseresponseafterP.aeruginosainfectionABCDHuangX.

etal.

(2006)IOVS47:4910-4916012345BALB/cTLR4lps-d1d3d5dDaDayspostinfectionLog10CFU/cornea3d5d024681012BALB/cTLR4lps-dp<0.001p<0.001DayspostinfectionUnitsMPO/cornea3d5d060120180240300BALB/cTLR4lps-dp<0.01p<0.001HuangX.etal.

(2006)IOVS47:4910-4916Bacterialload&MPOassayDayspostinfectionLog10CFU/coPro-inflammatorycytokineproductionHuangX.etal.(2006)IOVS47:4910-4916DayspostinfectionDayspostinfectionIL-1bprotein(pg/ml)3d5d03000600090001200015000BALB/cTLR4lps-dp<0.01p<0.001DayspostinfectionMIP-2protein(pg/ml)3d5d030006000900012000BALB/cTLR4lps-dp<0.001p<0.001RelativemRNAofIFN-g1d3d5d04080120160200240280BALB/cTLR4lps-dp<0.01p=0.35p<0.01DayspostinfectionRelativemRNAofIL-181d3d5d050100150200250BALB/cTLR4lps-dp<0.01p<0.01p<0.01Pro-inflammatorycytokineprodDayspostinfectionRelativemRNAofiNOs1d3d5d0816243240BALB/cTLR4lps-dp<0.05p<0.01p<0.05DayspostinfectionRelativemRNAofb-Defensin-21d3d5d01020304050BALB/cTLR4lps-dp<0.05p=0.83p<0.05iNOsandb-defensin-2productionHuangX.etal.(2006)IOVS47:4910-4916DayspostinfectionRelativemRNSummaryofTLR4inBacterialKeratitisIntheinfectedcornea:TLR4-deficiencyincreasescorneal inflammation;IncreasesTh1-typecytokineproduction;DecreasesiNOsandb-defensin-2 production;Reducesbacterialkilling.SummaryofTLR4inBacterialKTLR9inbacterialkeratitis

AkiraS.etal.(2005)Int.Immunol.17:1-14

TLR9inbacterialkeratitisAkBALB/cB601020304050601dayp.i.NormalATLR9mRNAlevelsTLR9IL-1bMIP-20102030405060GpCcontrolCpGtreatmentBRalativemRNAlevelsIL-12IL-18IFN-g0102030405060GpCcontrolCpGtreatmentCRelativemRNAlevelsIL-4IL-100102030405060CpGtreatmentGpCcontrolDRelativemRNAlevelsTLR9expressionandsignalingactivationinthecorneaHuangX.etal.(2005)IOVS46:4209-4216BALB/cB601020304050601dayp.

TLR9template

PCRreaction

T7-TLR9

TranscriptionTLR9dsRNA

RNaseIIITLR9siRNAATLR9temp

T7-TLR9

BTLR9dsRNACOverviewofTLR9-siRNAProductionHuangX.etal.(2005)IOVS46:4209-4216RNAinterference(RNAi):

anaturallyoccurringbiologicalprocessfor silencinggeneexpressionandfunctionbysiRNA.SmallinterferingRNA(siRNA):

asmallssRNAfragmentspecificallydegrade homologousRNAinmammaliancells.

TLR9templateATLR9tempTLR9-siRNAinhibitsmRNAexpressioninvitroHuangX.etal.(2005)IOVS46:4209-4216TLR9-siRNAinhibitsmRNAexpreTLR9silencinginvitro(Rho-CpGasareadout)HuangX.etal.(2005)IOVS46:4209-4216TLR9silencinginvitroHuangTLR9SilencingInVivo0123456Control-siRNATLR9-siRNA1d3d5dDayspostinfectionClinicalscoreCIL-12IL-18IFN-

g0102030405060Control-siRNATLR9-siRNARelativemRNAlevelsDIL-4IL-100102030405060TLR9-siRNAControl-siRNARelativemRNAlevelsEABTLR9-siRNAControl-siRNAHuangX.etal.(2005)IOVS46:4209-4216TLR9SilencingInVivo0123456CConfirmingSilencingWorksInvivoHuangX.etal.(2005)IOVS46:4209-4216ConfirmingSilencingWorksInIL-1bELISAControlTLR9-siRNA02000400060008000AIL-1b(pg/ml)MIP-2ELISAControlTLR9-siRNA0400080001200016000BMIP-2(pg/ml)ControlTLR9-siRNA050100150200250CUnitsMPO/corneaControlTLR9-siRNA0246810DLog10CFU/corneaELISAandMPOAnalysesHuangX.etal.(2005)IOVS46:4209-4216IL-1bELISAControlTLR9-siRNSummaryofTLR9dataIntheinfectedcornea,TLR9-siRNAtreatment:Reducesinflammation;Down-regulatesTh1-typecytokines;Down-regulatesPMNinfiltrate;Increasesbacterialload.SummaryofTLR9dataIntheinf科研課題：固有免疫識(shí)別與免疫應(yīng)答TLR9:

被細(xì)菌DNA激活

導(dǎo)致過(guò)度炎癥反應(yīng)角膜潰瘍的關(guān)鍵分子之一HuangXetal,

IOVS,2005;46:4209-4216（第一和通訊作者：黃曦）TLR4:

被內(nèi)毒素LPS激活免疫病理?yè)p害的分子基礎(chǔ)

角膜抗感染免疫所必需HuangXetal,

IOVS,2006;47:4910-4916（第一和通訊作者：黃曦）科研課題：固有免疫識(shí)別與免疫應(yīng)答TLR9:TLR4:NegativeregulationofTLRsignaling1st

SolubleTLRssuchassTLR2andsTLR4,actas decoyreceptorsforTLR2andTLR4signaling.2nd

TransmembraneproteinssuchasSIGIRRand ST2,sequesterrecruitmentofadaptorsandinhibit formationofTLRsignalingcomplex.3rd

IntracellularregulatorssuchasMyD88s,SOCS, IRAK-M.4th

ReductionofTLRexpressionorincreasedTLR degradation. 5th TLR-inducedapoptosis.6th TLRtolerance.NegativeregulationofTLRsigNegativeregulationofTLR4signaling

NegativeregulationofTLR4siSIGIRRandST2:MembersofimmunoglobulindomainsubgroupofTLR/IL-1Rsuper-familyNegativelyregulateTLRsignalingLigandunknown(SIGIRR)LigandforST2(IL-33)Lietal.(2005)J.Mol.Med.83:258-266SIGIRRandST2:Lietal.(200TLR負(fù)調(diào)控：抑制過(guò)度炎癥反應(yīng)ST2:

調(diào)控Th1/Th2細(xì)胞因子平衡

抑制TLR信號(hào)轉(zhuǎn)導(dǎo)通路

抑制免疫病理?yè)p害IOVS,2007;48:4626-4633（第一作者：黃曦）SIGIRR:直接抑制TLR4表達(dá)

角膜抗感染免疫所必需抑制免疫病理?yè)p害JImmunol.2006;177:548-556

（通訊作者：黃曦）TLR負(fù)調(diào)控：抑制過(guò)度炎癥反應(yīng)ST2:SIGIRR:

Roleof

SIGIRRinresistanceagainstP.aeruginosakeratitisRoleofSIGIRRinresistanceBacteriaPseudomonasaeruginosaATCCstrain196601x106CFU/ml(5ml)appliedtoscarifiedcorneaBacteriaPseudomonasaeruginosaAnimalmodelofP.aeruginosainfectionC57BL/6(B6)mice Susceptible(cornealperforationwithin5-7daysp.i.)BALB/cmice Resistant(restorecornealclaritywithin1-2wkp.i.)

AnimalmodelofP.aeruginosaN12h1d3d5d020406080100p<0.001p<0.001p<0.01p=0.23p=0.41DayspostinfectionSIGIRRmRNAlevelsBALB/cC57BL/6SIGIRRmRNAexpressionHuangX.etal.(2006)J.Immunol.177:548-556N12h1d3d5d020406080100p<0.0CornealdiseaseADayspostinfection012345

IgGcontrolAnti-SIGIRRAb1d3d5d7dClinicalscoreBCDEHuangX.etal.

(2006)J.Immunol.177:548-556CornealdiseaseADayspostinfec3d5d024681012

IgG

ControlAnti-SIGIRR

Abp<0.001p=0.007DayspostinfectionLog10

CFU/corneaBacterialloadHuangX.etal.

(2006)J.Immunol.177:548-5563d5d024681012IgGControlAnti-1d3d5d0200040006000800010000IgGcontrolAnti-SIGIRRAbp<0.001p<0.01Ap=0.26DayspostinfectionIL-1b

protein(pg/ml)1d3d5d030006000900012000150001800021000IgGcontrolAnti-SIGIRRAbp<0.001p<0.001p=0.03BDayspostinfection

MIP-2protein(pg/ml)Pro-inflammatorycytokineproductionHuangX.etal.(2006)J.Immunol.177:548-5561d3d5d0200040006000800010000I1d3d5d020406080100120IgGcontrolAnti-SIGIRRAbAP<0.001P<0.01P=0.12DayspostinfectionRelativeIL-1R1mRNAlevels1d3d5d0255075100IgGcontrolAnti-SIGIRRAbBP=0.016P=0.01P=0.67DayspostinfectionRelativeTLR4mRNAlevelsIL-1R1andTLR4expressionHuangX.etal.(2006)J.Immunol.177:548-5561d3d5d020406080100120IgGcont

1d3d5d020406080100120

IgGcontrolAnti-SIGIRRAbBp<0.05p<0.01p=0.58DayspostinfectionRelativeIFN-gmRNAlevels1d3d5d0255075100

IgGcontrolAnti-SIGIRRAbAp<0.01p<0.01p<0.01DayspostinfectionRelativeIL-18mRNAlevelsTh1-typecytokineproductionHuangX.etal.(2006)J.Immunol.177:548-5561d3d5d020406080100120IgGcon1d3d5d0255075100IgGcontrolAnti-SIGIRRAbAp=0.94p=0.86p=0.58DayspostinfectionRelativeIL-4mRNAlevels1d3d5d0255075100IgGcontrolAnti-SIGIRRAbBp=0.42p=0.89p=0.93DayspostinfectionRelativeIL-10mRNAlevelsTh2-typecytokineproductionHuangX.etal.(2006)J.Immunol.177:548-5561d3d5d0255075100IgGcontrolAnBSIGIRRmRNAexpressionNormal6h18h060120180240300LPSHK-PAASIGIRRmRNAlevelsEmptyvectorRAW264.7cellRAW264.7cellSIGIRR-vectorSIGIRR-vectorEmptyvectorC

SIGIRRproteinlevelsEmptyvectorEmptyvectorPositivecontrolSIGIRR-vectorSIGIRR-vectorSIGIRRSIGIRRexpressioninRAW264.7cellsHuangX.etal.(2006)J.Immunol.177:548-556BNormal6h18h06012018024030IL-1R1TLR-4TLR304080120160200EmptyvectorSIGIRRvectorAp<0.01p<0.001p=0.83RelativemRNAlevelsIL-12IL-18IFN-g050100150200250EmptyvectorSIGIRRvectorBp<0.01p<0.001p<0.01RelativemRNAlevelsSIGIRRinhibitsIL-1R1,TLR4andTh1-typecytokineproductionHuangX.etal.(2006)J.Immunol.177:548-556IL-1R1TLR-4TLR304080120160200EAIL-5IL-10020406080100Empty-vectorSIGIRR-vectorp=0.64p=0.78RelativemRNAlevelsIL-1bMIP-2100101102103EmptyvectorSIGIRRvectorBp<0.01p<0.01RelativemRNAlevelsCProteinlevels(pg/ml)IL-1bMIP-2100101102103104105106EmptyvectorSIGIRRvectorp<0.01p<0.01SIGIRRinhibitspro-inflammatorycytokines,butnotTh2-typecytokinesHuangX.etal.(2006)J.Immunol.177:548-556AIL-5IL-10020406080100Empty-veSummaryofSIGIRRIntheinfectedcornea,SIGIRR:IsdisparatelyexpressedinB6andBALB/c miceafterP.aeruginosainfection.Down-regulatesTh1,butnotTh2cytokine production;NegativelyregulatesIL-1RandTLR4expression andsignaling.SummaryofSIGIRRIntheinfectRoleofST2activationinP.aeruginosakeratitisRoleofST2activationHowtoactivateST2signaling?IL-33hasbeenidentifiedasaspecificligandofST2.WhetherornotIL-33iseffectiveinattenuatingocularinflammationthroughST2signaling?HowtoactivateST2signaling?RelativeST2mRNAlevelsN1350306090120150180C57BL/6BALB/cp<0.01p<0.05p<0.01p=0.26DayspostinfectionST2mRNAexpressionRelativeST2mRNAlevelsN13503ST2proteinexpressionRelativeST2proteinlevelsN350.00.51.01.52.0C57BL/6BALB/cp<0.01p<0.001p=0.35DayspostinfectionST2proteinexpressionRelativeIL-33treatmentofB6miceWhetherincreasinglevelsofIL-33(protein)inthecorneamodulatedP.aeruginosakeratitis.Todothis,B6mice(n=5/group/treatment)wereinjectedsubconjunctivallywith1mgIL-33protein(orPBScontrol)1daybeforeinfection.

IL-33treatmentofB6miceWhetAprotectiveroleofIL-33012345135PBSmIL-33p<0.01p<0.05p<0.01ADayspostinfectionClinicalscoreCBSlitLampat5daysp.i.PBSIL-33AprotectiveroleofIL-330123N150123mIL-33PBSp<0.01p<0.01p=0.77ADayspostinfectionRelativeIL-1bmRNAN1501234mIL-33PBSp<0.01p<0.01p=0.32BDayspostinfectionRelativeMIP-2mRNAN1501234mIL-33PBSp<0.05p<0.01p=0.13CDayspostinfection