分子醫(yī)學(xué)-炎癥、感染與相關(guān)疾病

分子醫(yī)學(xué)（MolecularMedicine）炎癥、感染與相關(guān)疾病InflammationInflammationispartofthecomplexbiologicalresponseofvasculartissuestoharmfulstimuli,suchaspathogens,damagedcells,orirritants.Inflammationisaprotectiveattemptbytheorganismtoremovetheinjuriousstimuliandtoinitiatethehealingprocess.FourMajorSymptomsofInflammation:1.Redness2.Pain3.Heat4.SwellingMayalsoobserve:5.LossoffunctionFunctionsofInflammationDestroyandremovepathogens(aswellasdamagedselftissuesandcells).Ifdestructionisnotpossible,tolimiteffectsbyconfiningthepathogenanditsproducts.Repairandreplacetissuedamagedbypathogenanditsproducts.StagesofInflammation1.Vasodilation:Increaseindiameterofbloodvessels.Triggeredbychemicalsreleasedbydamagedcells:histamine,kinins,prostaglandins,andleukotrienes(組胺，激肽，前列腺素，白三烯).2.PhagocyteMigrationandMargination:Marginationistheprocessinwhichphagocytessticktoliningofbloodvessels.Diapedesis(Emigration):Phagocytessqueezebetweenendothelialcellsofbloodvesselsandentersurroundingtissue.Phagocytesareattractedtositeofinfectionthroughchemotaxis.Phagocytesdestroymicrobes,aswellasdeadanddamagedhostcells.3.TissueRepair:Deadanddamagedcellsarereplaced.DefinitionofInfectiousDiseases

Alsoknownascommunicablediseases,ortransmissiblediseasescompriseclinicallyevidentillnessresultingfromtheinfection,presenceandgrowthofpathogenicbiologicalagentsinanindividualhostorganism.Infection感染Immunityisthebody’ssolutiontoprotectitselffromInfectiousDiseasesCommensalflora(共生菌群):staph,strep,pneumonia(葡萄球菌，鏈球菌，肺炎)Opportunisticpathogens(條件致病菌):candida,Clostridiumdifficile(念珠菌，艱難梭狀芽胞桿菌)Truepathogens(致病菌):InfluenzaContentsBriefintroductionofInfectiousDiseasesBacteriainfectionandInnateImmunityagainstbacteriainfectionViralinfectionandInnateImmunityagainstviralinfectionBriefintroductionofInfectiousDiseases1951年中國城市居民前10位死因:

肺炎;中樞神經(jīng)系之血管病變;胃炎,十二指腸炎,腸炎及大腸炎;心臟疾病;惡性腫瘤;周產(chǎn)期之死因;結(jié)核病;意外災(zāi)禍;自殺及自傷;腎炎及腎水腫InfectiousdiseasesisstillamajorprimarycausesofdeathworldwideMorensDMetal.,Nature2004,430:242-9.ManyCancersarealsorelatedtoviralinfectionEmergingandre-emerginginfectiousdiseasesMorensDMetal.,Nature2004,430:242-9.ViralEvolutionandPathogen-Hostrelationship:Evolutionarypressureonimmunity

Manyviruses,inparticularRNAviruses,haveshortgenerationtimesandrelativelyhighmutationrates.Thiselevatedmutationrateallowsvirusestoquicklyadapttochangesintheirhostenvironment.Viralevolutioncausesproblemsinthedevelopmentofsuccessfulvaccinesandantiviraldrugs,asresistantmutationsoftenappearwithinweeksormonthsafterthebeginningofthetreatment.InnateimmunityagainstPathogenicMicrobes

Bacteria:

Virus

FungiBacteriaFACTS:largenumbersofsingle-celledprokaryotemicroorganisms.2.Typicallyafewmicrometersinlength;3.Everywhereinearth:water,soil,aswellasinlivebodiesofplantsandanimals4.10millioninagramofsoil;amillionin1Lwater,5×1030onearth;10timescellnumbersinhumanflora(菌群).BacteriaInfectionBacteriaFACTS:5.Thevastmajorityofthebacteriainthebodyarerenderedharmlessbytheprotectiveeffectsoftheimmunesystem,andafewarebeneficial.6.Afewspeciesofbacteriaarepathogenicandcauseinfectiousdiseases,includingcholera,syphilis,anthrax,leprosyandbubonicplague(霍亂，梅毒，炭疽，麻風(fēng)病和鼠疫).Themostcommonfatalbacterialdiseasesarerespiratoryinfections,withturberculosisalonekillingabout2millionpeopleayear.7.Antibioticsareusedtotreatbacterialinfectionsandinagriculture,soantibioticresistanceisbecomingcommon.8.Inindustry,bacteriaareimportantinsewagetreatment,theproductionofcheeseandyogurtthroughfermentation,aswellasinbiotechnology,andthemanufactureofantibioticsandotherchemicals.Endotoxin:EndotoxinsaretoxinsassociatedwithcertainGram-negativebacteriawhicharestructuralmoleculesofthebacteriathatisrecognizedbytheimmunesystem.Theprototypicalexamplesofendotoxinarelipopolysaccharide(LPS).Exotoxin:Toxinsexcretedbyamicroorganism,includingbacteria,fungi,algaeandprotozoa.Anexotoxincancausedamagetothehostbydestroyingcellsordisruptingnormalcellularmetabolism.Theyarehighlypotentandcancausemajordamagetothehost.PathogenesisofBacteriaInfectionExotoxinsofBacteriaInfectionDiseasesfromBacteriaInfectionAntibiotics:Antibioticsisacompoundorsubstancethatkillsorslowsdownthegrowthofbacteria.ResistanceTheemergenceofresistanceofbacteriatoantibacterialdrugsisacommonphenomenon.

Emergenceofresistanceoftenreflectsevolutionaryprocessesthattakeplaceduringantibacterialdrugtherapy.Theantibacterialtreatmentmayselectforbacterialstrainswithphysiologicallyorgeneticallyenhancedcapacitytosurvivehighdosesofanti-bacterials.Undercertainconditions,itmayresultinpreferentialgrowthofresistantbacteriawhilegrowthofsusceptiblebacteriaisinhibitedbythedrug.Misuses:Thefirstruleofantibioticsistrynottousethem,andthesecondruleistrynottousetoomanyofthem.—PaulL.Marino,

TheICUBookBacteriaInfectionRecognitionOpsonization,phagocytosisDestructionMemoryInnateimmunity

Phagocytes Macrophages Dendriticcells Neutrophils Receptors Toll-likereceptors C-typelectinreceptors Cytosolic NODproteins RNAhelicases DNArecognition MediatorsInflamatorycytokinesAdaptiveimmunity Humoral Antibodies Cellular TcellsMicrobialrecognitionininnateimmunereceptorsInvertebrates/VertebratesVertebrates固有免疫適應(yīng)性免疫RecognitionofMicrobeproductsbyPAMPSPathogen-associatedmolecularpatterns,orPAMPs,aremoleculesassociatedwithgroupsofpathogensrecognizedbycellsoftheinnateimmunesystem.Thesemoleculescanbereferredtoassmallmolecularmotifsconservedwithinaclassofmicrobes.TheyarerecognizedbyToll-likereceptors(TLRs)andotherpatternrecognitionreceptors(PRRs)inbothplantsandanimals.BacterialLipopolysaccharide(LPS)onbacterialcellmembrane(Gram-)BbacterialflagelinPeptidoglycanandlipoteichoicacid(磷脂壁酸)fromGrampositivebacteriaUnmethylatedCpGDNA。。。PAMPsfrombacteriaToll-likereceptors(TLRs;transmembranereceptors)RIG-I-likereceptors(RLRs;cytoplasmicRNAhelicases)NOD-likereceptors(NLRs;cytoplasmicsensors)C-typelectinreceptors(CLRs;transmembranereceptors)Pattern

RecognitionReceptors（PRRs）Recognitionofpathogen-associatedmolecularpatternsbyPRRsInnateImmunitytoextracellularBacteriaO’Neill,LukeA.J.“Immunity’sEarly-WarningSystem”.ScientificAmerican,Jan(2005),38-45.TLRs:Toll-likeReceptorsdiacyl-triacyl-lipopeptide?；?；flagellin:鞭毛蛋白;LPS:脂多糖(Lipopolysaccharides)

DrosophilaTollIdentifiedaproteincalled“Toll”meaning“weird”HelpstheDrosophilaembryotodifferentiateitstopfromitsbottom

(Neuraltubedevelopment)/genomics/papers/drosophila.html

TollandInnerPartofHumanIL-1RisSimilarSearchingforproteinssimilartoTollShowscytoplasmicdomainofTollrelatedtothatofhIL-1RIdentityextendsfor135aaDidn’tmakesenseWhydoesaproteininvolvedinhumaninflammationlooklikeoneinvolvedinflyneuraltubedevelopment?FliesuseTolltoDefendfromFungiInfectedTl-deficientadultflieswithAspergillusfumigatusAllfliesdiedafter2-3daysFliesuseTolltodefendfromfungiThus,inDrosophila,TollseemstobeinvolvedinembryonicdevelopmentandadultimmunitySurvivalrateofadultDrosophilainfected

withAspergillusfumigatusinToll-FliesuseTolltoDefendfromFungiDrosophilahasnoadaptiveimmunesystemThereforeneedsarapidantimicrobialpeptideresponseTwodistinctpathwaystoactivateantimicrobialpeptidegenesinadultsMutationsinTollpathwayreducesurvivalafterfungalinfectionHumanTollDiscoveryIninsect,IL-1receptorandtheTollproteinareonlysimilarinthesegmentswithinthecellhumanproteinsthattotallyresembletoTollwerelaterdiscoveredHumanTollDiscoveryAlignmentofthesequencesofhumanandDrosophilaTollproteinsHomologyovertheentirelengthoftheproteinchainshTollgenemoststronglyexpressedinSpleenandPBL(peripheralbloodleukocytes)TLRRecognitionofMicrobialComponents

Bacteria

MicrobialCompomentsSpeciesTLRusage

LPSGram-negativebacteriaTLR4DiacyllipopeptidesMycoplasmaTLR6/TLR2TriacyllipopeptidesBacteriaandmycobacteriaTLR1/TLR2LTA(脂磷壁酸)GroupBStreptococcusTLR6/TLR2PG(肽聚糖)

Gram-positivebacteriaTLR2PorinsNeisseriaTLR2LipoarabinomannanMycobacteriaTLR2FlagellinFlagellatedbacteriaTLR5CpG-DNABacteriaandmycobacteriaTLR9NDUropathogenicbacteriaTLR11/nobel_prizes/medicine/laureates/2011/#LPSistheligandforTLR4LargemoleculesfoundinoutermembraneofGram-negativebacteriaComprisedofalipidandsaccharidecomponentHighlyimmunogenicRecognizedbyTLR4CancausesepticshockandleadtodeathOftenreferredtoasEndotoxinsinceitisnotsecretedbutisabyproductofbacteriallysisTLR4ActivatedbyLPSNormalmicedieofsepsisafterbeinginjectedwithLPSC3H/HeJmicehavedefectiveresponsetoLPSandsurviveMissensemutationaffectingthecytoplasmicdomainofTLR4Majorbreakthroughinthefieldofsepsis–molecularmechanismthatunderliesinflammationrevealedLPSLPSCellularlocationofTLRsTLRs:StructureLigandRecognitionTLR3-dsRNALigandRecognitionTLR4/MD2–LPSMyD88-dependentandindependentpathwayCytoplasmictailsofTLRsshowsimilaritiestoIL-1receptor(TIR)CommonadaptortoTLRsisMyD88CrucialprolineresidueinallTLRTIRdomains,exceptTLR3Ifmutatedordeleted,nosignalingoccursAllTLRslikelyhaveaMyD88pathway(TLR3isanexception)TLR4hasaMyD88independentpathwayaswellCytoplasmicTIRdomainTLRsandTIRDomainMyD88knockoutmicehavenoresponsetoLPSMyD88isessentialtoallinflammatorysignalingpathwaysMyD88s,asplicevariantofMyD88downregulatestheinflammatoryresponseMyD88interactswithTIRdomainofTLRandrecruitsIRAK-4,IRAK-1andTRAF-6MyD88AdaptorRakoff-Nahoumetal.,RecognitionofcommensalmicroflorabyToll-likereceptorsisrequiredforintestinalhomestasis.Cell118:229-41,2004.4IRAKsknown,IRAK1,IRAK-2,IRAK-MandIRAK-4IRAKareserine/threoninekinasesIRAK-4phosphorylatesIRAK-1IRAK-MplaysaninhibitoryroleinTLRsignalingTRAF6isamemberoftheTNFreceptorassociatedfactor(TRAF)familyTRAF6interactswithIRAK-1andgetsactivatedReleaseofTRAF6/IRAK-1ensuessubsequentsignalingIRAKandTRAF6IL-1RI-associatedproteinkinases(IRAKs)tumornecrosisfactorreceptor-associatedfactor6(TRAF6).TRAF6/IRAK-1complexassociateswith3proteinsTAK1(TGF-Bactivatedkinase)TAB1(TAK1bindingproteins)TAB2(TAK1bindingproteins)LargecomplexassociateswithmembraneEventuallyIRAK-1staysinmembranewhileTRAF6/TAK1/TAB1/TAB2movetocytosolE2LigasessuchasUbc13andUev1AjoinfurtherenlargingcomplexIRAKandTRAF6ReleaseTAK-1ActivationTheenlargedcomplexthatincludesTRAF6,TAK1,TAB1,TAB2,Ubc13,Uev1AactivateTAK1ActivatedTAK1phosphorylatesIKKcomplexActivatedTAK1canalsophosphorylateMAPKinasesIKKcomplexconsistsofIKK,and/NEMOIBphosphorylationresultsinNF-BtranslocationtonucleusOverviewofMyD88-dependentpathwayStepbyStepviewofMyD88-mediatedsignalingMyD88IndependentPathwayMyD88KnockoutmicedonotproduceinflammatorycytokinessuchasTNF-

HoweverwithTLR4stimulationNF-BandJNKdelayedactivityoccursThisstronglysuggeststheexistenceof2pathwaysinTLRsignalingaMyD88dependentpathwayaMyD88independentpathwayTLR3stimulationalsoexchibitsaMyD88independentpathwayTLR4OverviewofTLRsignalingInflammationCytokinesChemokinesFluidsProteinsBacteriatriggermacrophagestoreleasecytokinesandchemokinesVasodilationandincreasedvascularpremeabilitycauseredness,heat,andswellingInflammatorycellsmigrateintotissue,releasinginflammatorymediatorsthatcausepainInflammationmediatedbycytokinesLowmolecularweight,solubleproteinsthatareproducedinresponsetoanantigenandfunctionaschemicalmessengersforregulatingtheinnateandadaptiveimmunesystemInnateimmunesystemMacrophagesandDendriticcellsTumornecrosisfactor-alpha(TNF-)Interleukin-1(IL-1)Interleukin-12(IL-12)Adaptiveimmunesystem……………InflammationPro-inflammatorycytokines(TNF,IL-1)signaltoendothelialcellstomakethem:Leakytofluid(influxofplasma;containingantibodies,complementcomponents,etc.)Stickyforleukocytes,leadingtoinfluxofneutrophilsfirst,thenmonocytes,lymphocytesSystemiceffects:fever,acutephaseresponseSepsis(Septicshock)BacterialsepticemialeadstoactivationofTLRsonmonocytesinthebloodSystemicreleaseofTNFandIL-1leadsto“inflammation”alloverthebodyShockfromlossofbloodpressure(vasodilationandleakageoffluidintotissues)Thecombinationofeffectscanleadtomulti-organfailureanddeathBacteriaInfectionafterinjury—SepsisinEmergencyRoomNeonatalSepsisNeonatalSepsis？NegativeRegulationofToll-likereceptorsignalingSolubleTLRsMembrane-associatedRegulatorsFeedbackRegulationofIntracellularsignalingTLRscanalsosignalingtoapoptosisNLRTheNOD-likereceptors(NLRs)arecytoplasmicproteinsthathaveavarietyoffunctionsinregulationofinflammatoryandapoptoticresponses;TheofficialdefinitionofNLRiscurrently"Nucleotide-bindingdomain,Leucine-Richrepeatcontaining"proteinsincetheseproteinsarecomposedofconserved"modules"includingacentralnucleotide-bindingoligomerizationdomainandaseriesoftandemleucine-richrepeats.NLRsareencodedbyalargegenefamiliesinmanydifferentanimalspecies;therearemorethan20NLRgenesinhumans.Manyarethoughttoserveaspatternrecognitionreceptors(PRRs)whichsensemicrobialproductsinthecytoplasmofcells,althoughsomemembershavedifferentfunctions.NOD1&NOD2recognizepeptidoglycansubstructuresandpromoteinnateimmuneresponsesNOD1andNOD2areintracellularmoleculesandresemblesomeplantdiseaseresistanceproteins;bestunderstoodofthe“NOD-likereceptors”orNLRsNLR

signalingCARD

Domains：CaspaserecruitmentdomainsProcessingofIL-1andrelatedcytokinesbyinflammasome:animportantregulatorystepSome“NLRs”assembletoformthe“inflammasome”whichproteolyticallyprocessesIL-1andrelatedcytokinestotheiractive,secretedforms.InflammasomeinactivatedbycellularstressorrecognitionofmicrobialcomponentsinthecytoplasmGeneticperiodicfeversyndromesareduetoactivatingmutationsininflammasomeNOD-likereceptor(NLR)familymemberssuchasNALPs,NAIP,andIPAFTheLRRofNALP3orIPAFsensetheactivatingsignalsleadingtotheoligomerizationoftheNACHTregion.TheexposedPYDandCARDdomaincanrecruitadaptorproteinASCandCARD-containingcaspasestoformadonutshapecomplex.TheIL-1β-processingcaspaseactivitymostlikelyfacetheinsideofthedonut(lowerpanel).NACHTPYDCARDTwo

signalsareessentialforIL-1secretionInnateImmunityagainstViralinfectionViralinfection:Avirusisasmallinfectiousagentthatcanreplicateonlyinsidethelivingcellsoforganisms.EntrymediatedbyCellsurfaceReceptors:ViralInfectionVirus

receptor

celltypeHIV CD4 ThcellsEBV CR2 BcellsInfluenza sialicacid manycelltypesRhinovirus ICAM-1 manycelltypesPoliovirus poliovirusreceptor neuronsMeasles CD46 manycelltypesHHV6 CD46 manycelltypes GeneralProcessofViralInfection:ViralInfectionAttachmentPenetrationUncoatingReplicationself-assemblyReleaseCelldamagesmediatedbyviralinfection-EffectsonthehostcellMostvirusinfectionseventuallyresultinthedeathofthehostcell.Thecausesofdeathincludecelllysisandapoptosis。Oftencelldeathiscausedbycessationofitsnormalactivitiesbecauseofsuppressionbyvirus-specificproteinsSomevirusescausenoapparentchangestotheinfectedcell（latentinfection）.Thiscausespersistentinfectionsandthevirusisoftendormantformanymonthsoryears.Thisisoftenthecasewithherpesviruses.Someviruses,suchasEpstein-Barrvirus,cancausecellstoproliferatewithoutcausingmalignancy,whileothers,suchas,papillomaviruses(HPV乳頭狀瘤病毒)areestablishedcausesofcancer.ViralInfectionViralInfection

ViralNucleicAcidsrecognizedbyTLRsTLR3,TLR7andTLR8detectviralnucleicacidsFoundinintracellularmembranessinceviralnucleicacidsareendogenouslygeneratedViralNucleicAcidsrecognizedbyTLRsViralNucleicAcidsrecognizedbyTLRsRecognitionofcytosolicviralRNAsbyRLRsRIG-I-likereceptors(RLRs),alsoknownasRIG-I-likehelicases(RLHs)constituteafamilyofcytoplasmicRNAhelicasesthatarecriticalforhostantiviralresponses.

RIG-I(retinoic-acid-inducibleprotein1,alsoknownasDdx58)andMDA-5(melanoma-differentiation-associatedgene5,alsoknownasIfih1orHelicard)sensedouble-strandedRNA(dsRNA),areplicationintermediateforRNAviruses,leadingtoproductionoftypeIinterferons(IFNs)ininfectedcells.

RIG-IandMDA-5containaDExD/HboxRNAhelicaseandtwocaspaserecruitingdomain(CARD)-likedomains.ThehelicasedomaininteractswithdsRNA,whereastheCARDdomainsarerequiredtorelaythesignal.

RIG-IparticipatesintherecognitionofParamyxoviruses(Newcastlediseasevirus(NDV),Sendaivirus(SeV)),Rhabdoviruses(vesicularstomatitisvirus(VSV)),Flaviviruses(hepatitisC(HCV))andOrthomyxoviruses(Influenza).MDA-5isessentialfortherecognitionofPicornaviruses(encephalo-myocarditisvirus(EMCV))andpoly(I:C),asyntheticanalogofviraldsRNA.

Notably,RIG-IbindsspecificallytosinglestrandedRNAcontaining5’-triphosphatesuchasviralRNAandinvitro-transcribedlongdsRNA[4].MammalianRNAiseithercappedorcontainsbasemodificationssuggestingthatRIG-Iisabletodiscriminatebetweenselfandnon-selfRNA.RIG-IbindspreferentiallytoshortdsRNAwhileMDA-5recognizespreferentiallylongdsRNA.ViraldsRNAisalsorecognizedbyToll-Likereceptor3(TLR3)whichisexpressedonthecellsurfacemembraneorendosomes.

RecognitionofdsRNAbyRIG-I/MDA-5orTLR3iscell-typedependent.StudiesofRIG-I-andMDA-5-deficientmicehaverevealedthatconventionaldendriticcells(DCs),macrophagesandfibroblastsisolatedfromthesemicehaveimpairedIFNinductionafterRNAvirusinfection,whileproductionofIFNisstillobservedinplasmacytoidDCs(pDCs).ThusincDCs,macrophagesandfibroblasts,RLRsarethemajorsensorsforviralinfection,whileinpDCs,TLRsplayamor