Download presentation
1 -

Tuberculosis2006273


TuberculosisRUTIAnewchancetoshortenthetreatmentoflatenttuberculosisinfectionPere-JoanCardonaUnitatdeTuberculosiExperimentalDepartmentofMicrobiologyFundacioInstitutperalaInvestigacioenTel34934978894fax

oneill's Recent Documents

x0000x0000  1  xMCIxD 0 xMCIxD 0  xMCIxD 1 xMCIxD 1 FEEDBACK REG
x0000x0000 1 xMCIxD 0 xMCIxD 0 xMCIxD 1 xMCIxD 1 FEEDBACK REG

Masic I Miokovic M Muhamedagic B Evidence based medicine new approaches and challengesActa Inform Med 2008164219RITISH COLUMBIA HUMANIST ASSOCIATION422 Richards St Suite 170Vancouver BC V6B 2Z4 604 26

published 0K
PROFESSIONAL CONGRESS ORGANISER
PROFESSIONAL CONGRESS ORGANISER

Eethnopharmacology2021artioncomgrT 30 2310257801 congress line 30 2310272275wwwartioncomgronal ongof onal SocogVirtual Congresswwwethnopharmacology2021orgISEonal ongof onal SocogVirtual CongressA case

published 0K
273  Other licenses
273 Other licenses

GS 113-273Page 1 113-aExcept when otherwise indicated licenses in this section are annual licenses issued beginning January 1 each year running until the following December 31bExcept as otherwise prov

published 0K
JournalofMachineLearningResearch15201432973331Submitted913Revised51
JournalofMachineLearningResearch15201432973331Submitted913Revised51

TanLondonMohanLeeFazelandWitten1IntroductionGraphicalmodelsareusedtomodelawidevarietyofsystemssuchasgeneregulatorynetworksandsocialinteractionnetworksAgraphconsistsofasetofpnodeseachrep-resentingavari

published 0K
what_about_writing_.pdf
what_about_writing_.pdf

0021306-70084480029125-0122

published 0K
1Fx008ex008e3x008bte6x008bsx008bt2eport0repredbx009bCffx008bceofx000cn
1Fx008ex008e3x008bte6x008bsx008bt2eport0repredbx009bCffx008bceofx000cn

Armijo6x001b00onalesRdSJx000fAlNuquerquex000fNMx001bx001a121ScOoolIeadersOipMonicaAguilarx000fEecutiQeGirectoroQerningBoardMemNersMelissaArmiMox000fPresident-acoNomex000fVice-PresidentCOarlotteAlderet

published 0K
Documento descargado de httpzlelsevieres el 11112014 Copia para us
Documento descargado de httpzlelsevieres el 11112014 Copia para us

Documento descargado de http//zlelsevieres el 11/11/2014 Copia para uso personal se prohbe la transmisin de este documento por cualquier medio o formatoDocumento descargado de http//zlelsevieres el 11

published 0K
PRESSFIT BUTTONS
PRESSFIT BUTTONS

STANDARD FLAT LOCATION IS AT 0 AS SHOWN AVAILABLE AT 90 180 270 FOR SAME ALTERATION PRICEMD- 00 B R D L 50 15 n 5 10-25 j 6 32-100 RP or G 8mm/32 max 180PGWGWPWRPRWPRROUNDMDCOBLONGMDOSQUAREMDSR

published 0K
Download Section

Download - The PPT/PDF document "" is the property of its rightful owner. Permission is granted to download and print the materials on this web site for personal, non-commercial use only, and to display it on your personal computer provided you do not modify the materials and that you retain all copyright notices contained in the materials. By downloading content from our website, you accept the terms of this agreement.






Document on Subject : "Tuberculosis2006273"‚ÄĒ Transcript:

1 Tuberculosis(2006),273 Tuberculosis RUTI
Tuberculosis(2006),273 Tuberculosis RUTI:AnewchancetoshortenthetreatmentoflatenttuberculosisinfectionPere-JoanCardonaUnitatdeTuberculosiExperimental,DepartmentofMicrobiology,FundacioīInstitutperalaInvestigacioīen Tel.:+34934978894;fax:+34934978895.E-mailaddress:pcardona@ns.hugtip.scs.es. Furtherexperimentsusingbiggeranimals(goatsandmini-pigs)willprovidemoredataontheefřcacyofRUTIbeforestartingphaseIclinicaltrials.2006ElsevierLtd.Allrightsreserved.Introduction:whatisalatentbacillus?Despitethisbeingasimplequestion,řndinganappropriateansweriscrucialtodemonstratethevalidityofanynewtreatmentagainstlatentMycobacteriumtuberculosisbacilli;latentbacilliareresponsibleforthelongperiodoftreatmentcurrentlyrequiredforsterilizinglesionsinfectedM.tuberculosisThenatureoflatentbacilliandtheirabilitytosurvive‘‘invitro’’inlowconditions,andeveninanaerobiosis,hasbeenwidelystudied.Tosurviveinlowbacilliappeartoacquireastateofnonreplicantpersistence(NRP).Hugelesionswithintragranulomatousnecrosis(IN)thatareinducedintuberculosis(TB)maydevelopsuchanextremeanaerobicenvironment,supportingtheideathatlatencyinducedbylowoccursinlesions.However,directmeasurementshaveneverdemonstratedthepresenceofanaerobiosisinTBlesionsandfactualsupportforthehypothesisislacking.Furthermore,INisa‘‘living’’tissue,fullofcollagenřbers,thatalsoneedsaphysiologicalenvironmentforsurvival.Anaerobiosis,therefore,appearsunlikelyinsuchasetting.Evenifsuchatmospherewithlowbeinducedincalciředlesions,experimentalmodelsdemonstratethattheperiodinwhichthebacillimaysurviveinthisenvironmentisdeřnitelyřnite.Moreover,micro-aerobiosisisusualinthehosttissuesand,there-fore,notonlylatentbacillibutalsoactivelygrowingcellsmayadapttothisenvironment.Furthermore,experimentsinknockout(KO)micelackingfunctionalgenesessentialfortriggeringacompetentimmuneresponsedemonstratedthatactivegrowingbacillicanbefoundinsidemassivelesionswithabundantIN,expectedtohavealowSomeauthorsdeřnedlatentbacilliasthosepresentinthetissuesofmicetreatedwithchemotherapyforalongtime,whosepresencewasnotdetectedinculturesuntilafewmonthsaftertheendofthetreatment;thisprocesswasfavoredbytheadministrationofcortisone.Thisexperimentalmodel,characterizedalongtimeago,isknownastheCornellmodel.IntheCornellmodelbacillidonothavetoresistanextremelylow.However,patientswithlatenttuberculosisinfection(LTBI)facecircumstancesdifferentfromthoseintheCornellmodel.Usually,peoplewithLTBIdisplayacell-mediatedandantibody-mediatedimmuneresponsethatcontrolsprogres-sionoftheinfectionintheinitialfocusofinfectionandinthelocaldraininglymphnodes(the‘‘Ghoncomplex’’)byinducingastronggranulomatousinřltration,aprocesswhichdoesnothappenintheCornellmodel.Besides,ItiscurrentlyacceptedthatthepopulationobtainedwiththeCornellmodelisspecialandisnotconsideredtobeagoodmodeloflatentbacilli,butratherof‘‘persistent’’bacilli.Persistenceimpliesthataspecialpopulationofbacillisubsiststheperiodofche-motherapy,probablythroughacquiringtolerancetoantibiotics,althoughitsmechanismremainspoorlyunderstood.Otherauthorsfocusedonthehypothesisthatlatentbacillihavetoadapttoalackofnutrients,thusresemblingthebacterialpopulationfoundinthesteadystateofconventionalliquidcultures.Thustheterm‘‘dormant’’wasestablishedtodeřnebacillithatwere‘‘inastateoflowmetabolicactivityandunabletodivideortoformacolonywithoutaprecedingresuscitationinliquidmedium,whichmayeitheroccurspontaneouslyorrequiretheprovisionofcompounds(growthfactors)pre-sentinthesupernatantofgrowingcells’’.13,14Theseauthorsconsideredtheinvitro‘‘dormant’’formastheequivale

2 nttotheoneobtainedinvivowiththeCornellmo
nttotheoneobtainedinvivowiththeCornellmodel,representinganextremeformofviability,closetothedeathofthecell.However,theexquisitelyfastidiousrequirementsneededforregrowthofthese‘‘dormant’’bacilliaredifřculttoreconcilewiththeconditionsfacedbylatentbacilliinvivo,bacilliwhichare,however,abletoreactivateafteralongperiodoftimetoinduceTBdisease.Finally,thereistheobservationmadealongtimeagothatthebacillarypopulationobtainedfromthelungsofchronicallyinfectedmicehasagreaterresistancetoheatstress(53C)thanthebacillarypopulationintheacutephase.Thesameauthorsubmitted‘‘invitro’’cultures,young(exponentialphase)andold(steady-statephase)tothesameconditionsandobtainedahigherresistancetoheatstressinthelatter.Thisobservationledtotheconclusionthataslowergrowthratemadebacillifromthechronicphasemoreresistanttostress.Consistentwiththishypothesis,Munųoz-Eliasetal.ARTICLEINPRESS P.-J.Cardona recentlyreviewedthisworkanddemonstratedthatthechronicphaseofinfectioninmurineTBwasaccompaniedbyareductionintherateofbacterialcell-division.ThecurrentconsensusisthatthechronicphaseintheexperimentalmurinemodelisaconsequenceofthespeciřcimmunityelicitedagainstgrowingM.tuberculosisThisimmunitytriggersanumberofmechanismsthatinducesthedeathofmostbacterialcells(i.e.lowpH,reactiveoxygenintermediates,reactivenitrogenintermediates,etc.).Asaconsequenceofthisstrongimmuneresponse,thepopulationinthechronicphasecomesfrombacilliabletoadapttostressandthus,ahighproportionofthesebacilliarethesameasso-called‘‘latent’’bacilli.Therefore,inouropinionlatentbacilliarethosethatcanresisttheim-munologicalresponsetriggeredbythehost.How-ever,itisnoteworthythatnotallbacillifromthechronicphaseareinalatentstate,consistentwiththeobservationsinpatientswithLTBI.ImmunopathologyofthescenariooflocalAtthebeginningofM.tuberculosisinfection,thedevelopmentofINsuggeststheoccurrenceoftheKochphenomenon,observedinmostmammals.TheKochphenomenonprobablyarisesfromalocalSchwartzmanreaction.19,20Thefollowingsequenceofeventsarepostulated:infectedmacrophagesinthegranulomaproducehighconcentrationsofTNF;thepresenceofthiscytokinemakesthesiteresponsivetoendotoxin-likemolecules(liketreha-losedimycolate),whichexistinthecellwallofM.tuberculosiswhenthelackofspeciřcimmunityallowsanexponentialgrowthofbacilli;thecorrespondingreactiontriggersa‘‘řrstwave’’oflatentbacilli(i.e.,bacillisurvivingtheinitialinŖammatoryresponse),whicharethentrappedinthecollagenřbersthat‘‘taketheplace’’ofthedestroyedinfectedmacrophages.A‘‘secondwave’’oflatentbacillimaybetriggeredbytheinductionofspeciřcimmunity,whichactivatestheinfectedmacrophagestodestroymostoftheinitialbacillarybulk.Thisresponsetospeciřcimmunityisobservedintheexperimentalmurinemodelofaerosol-inducedTB,whereINisnotusuallyinducedandwhereanti-body-mediatedimmunityisnottriggeredduringthisinitialphaseofinfection.Inthismodel,a90%reductionofthebacillaryconcentrationiscausedbytheactivationofinfectedmacrophagesbyTcellsthatproduceIFN-andbythecytolysisinducedbyspeciřcCD8cells..However,duringthechronicphase,newcellsappearattheoutermostlayerofthegranulomas,occupyingthealveolarspaces:thesecellsarethefoamymacrophagesFMareusuallyseeninallinŖammatoryresponsesinthelung,andnotonlyininfectiousinŖammatoryresponses.ThefunctionofFMistophagocytethenecroticmaterialproducedafteraninterstitialinŖammatoryresponse(i.e.deadhostcells,thesurfactantsecretedlocallyandthedeadM.tuberculosiscellsremaining),torebuildtheoriginalparenchyma.Thesenewlyaccumulatedmacrophagescanleavethelesionthroughthealveolarspacestoreachtheupperbronchi,wheretheya

3 reswallowedandenterthestomach.Intheconte
reswallowedandenterthestomach.InthecontextofM.tuberculosisinfection,theFMmaybealsooriginatedfromtheinitialinfectedmacro-phagepopulation,followingthekillingofmostofbacilli.Theaerosolmurinemodelpresentsaninterestingscenariobecauselargequantitiesofbacilliareobservedingranulomasthatareinitiallyill-deřned,duringtheacutephaseofinfection,whereasnoacidfastbacilliareseenintheircenterduringthechronicphase(at6weekspostinfec-tion);instead,singlebacilliarepresentintheperiphery,insideFMlocatedinthealveolarspaces.Furthercharacterizationofthesestructuresde-monstratedaprogressiveincreaseofinduciblenitricoxide(NO)synthase(iNOs)duringthechronicphaseofinfection,whichisthemainenzymeresponsibleforNOproductionandwhichisloca-lizedalmostexclusivelyinFM.Hence,sinceFMproduceNOandmaysuppresstheactivatingspeciřcTcells,bothTh1andTh2,theyconstituteanimmunosuppressiveoutermostringaroundthelesion.IntheacutephaseofM.tuberculosisproductionofRNIislinkedtoaTh1activation,andisparamountforthecontroloftheinfectionincludingthecontrolofchronicinfectioninthemurinemodelofTB.However,atthesametime,NOproduction,whichisstimulatednotonlybyIFN-butalsoTNFandIL-4inhumanmonocytesandpulmonaryepithelialcells,alsogeneratesanegativefeedbackintheproductionofthesecytokinesthusgeneratingalocalimmunosuppres-sionbothforTh1andTh2responses.Theanti-inŖammatoryroleofNOhasbeensuggestedbytakingintoaccountthatRNIarealsoextremelytoxictothehostcellsandnotallofthemareabletokillM.tuberculosis,(forexample,peroxynitriteanion,which,atthesametimeexertsanegativefeedbackagainstNOproduction).FMhavebeenextensivelystudiedintheinduc-tionofatherosclerosis.Inthiscase,theiroriginisARTICLEINPRESS RUTI:treatmentoflatenttuberculosisinfection275 linkedtoaninŖammatoryprocess.Inparticular,ithasbeendemonstratedthatTNFisabletoupregulatethelectin-likeoxidizedLDLreceptor(LOX-1)whichincreasestheuptakeofOxidizedLDL(OxLDL)thuscontributingtolipidosisandFMformation.Inthiscase,theincreaseofLOX-1waslinkedtoaNOdeřciency.Thus,thepresenceofNOdoesnotfavortheformationofFM.Ontheotherhand,ithasbeenrecentlydemonstratedthatoxidationofLDLisnotalimitingfactorforinductionofFMastheycanbealsoformedwithnativeLDLthroughamechanismofmacropino-cytosis.AdditionalevidenceinsupportoftheviewthatinŖammatoryresponseisparamountintheinductionofFMcomesfromthestudieswithIL-10KOmice,whichexhibitedathreefoldincreaseincholesterolaccumulationinmacrophages,whereasover-expressionofIL-10ineitherC57BL/6ormiceresultedinasigniřcantdecrease.Furthermore,treatmentofmicewiththenonsteroidalanti-inŖammatorydrugindomethacinoraCOX-2-selectiveinhibitor,rofexocib,resultedinasigniřcantreductioninlesionsize.Inthecontextofthelunginfectionitmustbetakenintoaccountthatthepresenceoflargeamountsofalipoproteinsurfactantcomplexmayalsoplayanimportantrole.Thename‘‘surfactant’’isderivedfromitswell-deřnedabilitytolowersurfacetensionattheinterfaceofalveolargasandtheliquidhypophase,andalsoparticipateinthecontrolofparenchymadestruction.Inthisregard,whenalveolarmacrophagesarechronicallyex-posedtohighquantitiesofsurfactanttheybecomeThepresenceofkilledM.tuberculosisorcellwallfragmentsfromthemalsoinducestheformationofFM.Furthermore,somecomponentsM.tuberculosisarealsoknowntoinduceiNOSproduction.ThismayexplainwhyapparentlyincoherentcircumstancesmaycoexistinthechronicphaseofM.tuberculosisinfectioninmice,i.e.thepresenceofFMinthecontextofNOexpressionandthecontinuedlocalimmunosuppres-sionoveralongperiod.Besides,asNOproductionintheFMmayoccurinthecontextofinsufřcientmacrophageactivation,asaconsequenceofthesuppressio

4 nofTh1response,thiscouldexplainwhythebac
nofTh1response,thiscouldexplainwhythebacillithatshelterinthemcansurvive.ItisknownthatRNIbactericidaleffectisconcentrationdependent.Thus,paradoxically,RNIareknowntobeabletoeveninducemycobacterialgrowth,andhavebeenimpliedinthehormesisphenomenon,i.e.stimulatoryeffectscausedbylowlevelsofapotentiallytoxicagent.Recently,severalreportshaveshownthatlowornontoxicconcentrationsofNOdonors(DETA-NO)modulatetheexpressionofa48-generegulon,whichisexpressedbyM.tuberculosisbothinvivoandinvitroandpreparesbacilliforsurvivalduringlongperiodsofinvivodormancy.Inconjunctionwithlowconcentrationsofoxygenintissues,NOcanplayaroleintheinitiationandmaintenanceofthelatentstateofM.tuberculosiswithingranulo-mas,whichcouldbeamechanismforthetuberclebacillustopersistinFMofthehost.addition,Darwinetal.haveshownthatmutationoftheproteasomeofM.tuberculosisbacillitoNO,suggestingthatmycobacterialpro-teasomeservesasadefenseagainstnitrosativeFMalsoproducelargequantitiesoftransforminggrowthfactor-beta(TGF-)andthusmayinduceapoptosisofeffectorTcells.TheyareknownalsotodisplayhighlevelsofDEC-205,(i.e.,amarkercharacteristicofdendriticcells),butdownregulateMHCclassIImarkers,thusrenderingtheminade-quateforantigenpresentation(althoughnodataonMHCClassImarkersareavailable).Besides,FMappeartoresistapoptosis.Thus,FMappeartobeakindofsanctuaryforthosebacillithatsurvivetheimmunologicalresponse,inadditiontoalsobeingasourceoflocalimmunosuppression.Furthermore,wemustalsoconsiderthehypothesisthatFMcannotproducematureendosomesandinducestressconditionsduetohavingtoomuchmaterialtoprocess.Thisinturn,mayencouragebacillarygrowthattheperipheryofthegranuloma.Finally,ithasbeenreportedthattreatmentwithisoniazid(INH)andrifampicinresultsintheeliminationoftheFMoutermostlayer,butitisstillnotclearifthisphenomenonislinkedtotheinhibitionoftheregrowthofthebacilliandthusthereductionoftheinŖammatoryresponse,orbythedirectreductionoftheinŖammatoryresponsecausedbytheantibiotic,asdemonstratedformoxiŖoxacyn.Figure1representsatime-relateddescriptionofthemurinemodelofTBwithregardstogrowing/deadbacilli,synthesisofsurfactant,IFN-andTNF,andFM.Allthesedataarebasedonpublishedwork.However,thekineticsofsurfactanthasbeeninterpretedbasedonitsfunction,asacomponentoftheinnateimmunityandconsideringitsroleagainsttissuedestruction.ThepeakofsurfactantoccursbeforethepeakofIFN-andTNFanditsproductionissustainedasaconsequenceofcumulativeinřltrationofthelungparenchy-causingconstantinjurytothistissue.Inthismodel,thepresenceofFMisaconsequenceoftheaccumulationofparenchymaldebris,surfac-tantanddeadM.tuberculosisNotethatiflatentbacilliremaininthemacro-phageinitiallyinfected,whichhasbeenactivatedARTICLEINPRESS P.-J.Cardona andhasdestroyedmostbacilli,anditbecomesaFM,itisreasonabletoassumethat,withtime,lowpHwillbeneutralizedandthebacilliwillre-growwhenFMareattheperipheryofthegranuloma.However,iftheseinfectedmacrophagesbecomeapoptoticornecrotizedinthecenterofthegranuloma,whathappenswhenthelatentbacilliisphagocytedbynewmacrophages?Thesebacilliaremetabolicallyinactiveandtheycannotavoidthephagosome-lysosomefusion,asitrequiresastrongmetaboliceffort.Theselatentbacilliarethenconsidered‘‘deadbacilli’’andmustface,onceagain,alowpHthatwillfurtherprolongtheirlatentstateuntilstressfulconditionsdisappear.ThishypothesismayalsoexplainwhyveryfewbacilliremaininFMandwhysofewaredrainedtotheperipherywheretheyřnallyre-grow.Interest-ingly,inbothsituationstheantigensofthelatentbacillicanbeeasilyprocessedduetheir‘‘deadlike’’status,andiftheyarepresentedinsideapoptoticvesicles,theyw

5 ouldenhanceantigenpresentationthroughMHC
ouldenhanceantigenpresentationthroughMHCIandCD1molecules.Paradoxically,latentbacillimighttriggeraspeciřcimmunity.Whydoesthisnothappen?Theanswermaybe‘‘duetolocalimmunosuppressioninducedbyFM’’or‘‘becausethemacrophagesharboringthemdonotreachthelymphnodestopresenttheantigens,asthisisnottheirrole’’.Finally,wemustalsoconsiderthehypothesisthatFMcannotproducematureendosomesandinducestresstobacilli,duetohavingtoomuchmaterialtoprocess,thusencouragingbacillaryregrowthattheperiphery.Infact,thepresenceoftheseFMiscrucialinthemurinemodel,astheyallowcontinuousinřltrationofthepulmonaryparenchymaduringthechronicphaseoftheinfection.However,thisphenom-enonseemstobelimitedinbiggerhosts,suchasguineapigsorhumans,wherethehosttriggersastronginŖammatoryresponseatthebeginningoftheinfection,inducingINandquicklysurroundingthegranulomawithařbroticmantle.Neverthe-less,thesefoamycellshavealsobeenobservedinsidethegranulomaharboringsinglebacilli.Allthesedatasupportthehypothesisthatlatentinfectioninhumansissustainedbyastaticpopulationinthenecrotictissuesandbyaconstantreactivationoflatentbacilliphagocytedbymacro-phagestryingtoreabsorbthenecrotictissues.M.tuberculosisrecognizestheabsenceofstressfulconditionsbymodulatingtheirgene-expressiondependingonexternalconditions(i.e.tionistriggeredwithacidpH),andmustalsodetectwhentheadverseconditionsdisappear(i.e.whenphagocytedbyFM)andthuswhentheycanstartgrowing.TheverylowmetabolicactivityofM.tubercu-alsofavorstheirsurvival.Thesebacillimayreactivatefarfromtheinitialinfectionfocus,awayfromthepresenceofeffectorTcells,andinsidethealveolarspacewhere,aftergrowinganddestroyingtheinfectedmacrophages,theycaneasilydisse-minatetobephagocytedbyothernaű¨vealveolarmacrophagesandreinitiateinfection.Thefocusoftheimmunologicalresponseisongrowingbacilli,whichpoolisconstantlyregenerated;incontrast,aresponseagainstlatentbacilli,ifitisevertriggered,remainsrelativelyweak.AsrepresentedinFig.2A,thesuggestedscenarioofLTBIincludesastaticpopulationoflatentbacillithatperiodicallyre-growsatdifferentsitesoftheparenchyma.Forabacillusinthiscontext,reachingaconvenientsiteforgrowthiscrucial(e.g.,theapicalzonesofthelungwithhighoxygenpressuretoinducealargegranulomaandacavern).ARTICLEINPRESS 0369121518 0246 024 relative values 0246 026 growing bacillifoamymacrophages 0246 surfactant Figure1EvolutionmodelinexperimentalmurineTB,focusingonthelocalpresenceofgrowing/deadbacilli;surfactant,IFN-andTNFproduction;andpresenceofFM.Continuousanddashedlinesmarkthepeakandthebaseofgrowingbacilli.Dottedlinesrepresentacycleintheevolutioninthechronicphase. RUTI:treatmentoflatenttuberculosisinfection277 Asaconsequenceofalltheabove,thestrategyofinducingastrongTh1responseisnotrelevantfordesigninganimmunotherapeuticstrategyagainstLTBI,especiallyifwetakeintoaccountthathumans,whoarethehoststhatmanagetocontrolM.tuberculosisinfectionmosteffectively,triggeraTh2response.Th1responsekillsgrowingbacilliwhereasTh2‘‘wallsoff’’latentbacilli,thelatterbeingresponsibleforgranulomařbrosis,togetherwiththepresenceofTGF-aTh3response.However,ithasbeensuggestedthatTGF-istheonlyfactorrequiredforřbrosingthegranulomas,i.e.toensurethatlatentbacillidonotescape.Inthisscenarioalso,a‘‘pure’’Th1response,suchasoccursinmice,doesnot‘‘close’’thegranulomasufřcientlyandthereforeallowsconstantdisseminationoftheinfection.more,asNOlocalproductionisparamountinchronicphasegranulomasandinhibitsbothTh1andTh2responsesitisnonsensetoincreaseanyofthemasitwillbeuseless.Interestingly,histopathologystudiesonhumanTBlesionshavedemonstratedthat

6 thepresenceofIL-4maynotbeanindicatorofpo
thepresenceofIL-4maynotbeanindicatorofpoorprognosisinsuchpatientsbutrathermaybeanintegralfeatureoftuberculousgranulomaformationwitharoleincontrollingtissuedamage.Inthesamestudy,thosepatientsexhibitingthehighestpercentageofTNF-positivegranulomasalsohaveIL-4-positivegranulomasandalowerpercentageofgranulomaswithcaseousnecrosis.WhydoesconventionalchemotherapyofLTBIneedsuchalongperiodofAccordingtotheřeld-workcarriedoutbyCom-thegold-standardtreatmentforLTBIistheadministrationofINHfor9months.Interestingly,thestudyofearlybactericidalactivity(EBA)ofINHinpatientswithpulmonaryTBshowsthatthehighestbactericidalactivityisachievedwithintheřrst2daysoftreatment(witha0.7logreduc-Essentially,thehighestbactericidaleffectofINHoccurswithintheřrst15dayspostadministration.Therefore,whyissuchalongtreatmentperiodrequired?ThisisbecauseINH,asmostbactericidaldrugs,isonlyeffectiveagainstactivelygrowingbacilli.Infact,inthecaseofcavitaryTB,INHiseffectivebecausethereisanimportantextracellularpopulationwithmanyactivelygrowingbacilli.ThelongperiodofchemotherapyrequiredinLTBItreatmentmaybeexplainedbytheabilityofchemotherapytostopconstantregrowthofbacilliFig.2B):thelackofgrowingbacillireduceslocalinŖammatoryandimmunologicalresponsesallowingreabsorptionofthenecroticdebris,surfactantanddeadbacilli,thussigniřcantlyreducingthepresenceofFMandlatentbacilli.Consideringthatthisdecreaseinlocalimmuneresponsesfacilitatesfuturere-growingoflatentbacilliifthesebacilliarenotcompletelyremoved,ARTICLEINPRESS 16 necrosis population 050100150200250300350400450500 146 days Figure2MathematicalapproachtolatencyinTBbasedontheconstantreactivationoflatentbacilli.Thereisanacutephasewherebacillireachmaximumconcentrationafter20daysofconstantgrowththatlastsuntiltheimmuneresponseistriggeredand99%ofthebacillarybulkisdestroyed(A).Thisreactionoriginatesfrom0.1%oflatentbacilliinnecrotictissue.Theremaining1%ofbacilliinmacrophagesre-growafterarestingperiodof20days,whenonceagain99%aredestroyed,originatinganother0.1%oflatentbacilliinanecrotictissue.Thisdynamiccontinuesuntilequilibriumisreachedatthetolerancethresholdinhumansof2log.Thenumberofbacilliinnecrotictissuedecreaseswhiletherestingperiodbetweenreactivationsincreasesproportionallytohistoricaldataofuntreatedtuberculosis-infectedhouseholdcontacts.Thisdatasuggeststhatthediseaseoccursatarateof0.74%peryearinyears1and2;0.31%peryearinyears35;and0.16%peryearinyears6and7.(B)showstheendofconstantre-growingwithche-motherapyandtheeliminationoflatentbacillifromnecrotictissuecalculatedinrelationtotheprotectionobtainedwithseveralperiodsoftreatmentwithINH.Thenumberofbacilliinsidemacrophagesisindicatedinred,whereasthebacillarynumberinnecrotictissuesisingray.Thetimeofchemotherapyispresentedinorange. P.-J.Cardona itiscrucialtomaintainlongchemotherapyperiodssoastoallowtotalremovaloflatentbacillifromthepulmonaryparenchyma.WhatdoesRUTIdo?ThedesignofRUTIRUTIwasdesignedtořlltheimmunologicalgapleftbyshort-termtherapyforM.tuberculosistion.Infact,ourgroupdemonstratedinpreviousinvestigationsandindifferentexperimentalmodelsofTBinmicethatM.tuberculosisgrewagainaftershort-termchemotherapy,althoughthiswasnotfollowedbyanimmediatestimulationofimmunity.Moreover,thisdelayedimmunestimulationwasonlytriggeredwhenthebacterialloadreachedavalueof4log(afteraperiodof2months),whichhadclearimplicationsforthe‘‘oldissue’’of‘‘tolerance’’ofmicetolargerbacillaryexposurethanhumans,andtheneedformorebacillitoboostimmunity.TheoriginalideaonintroducingRUTI,therefore,wasto‘‘boost’’theimmunologicalresponseagainstM

7 .tuberculosisthatalreadyexistedinthehost
.tuberculosisthatalreadyexistedinthehost.RUTIwasalsodesignedtotriggeranewimmunologicalresponseagainstantigensofthelatentbacilli,i.e.theso-called‘‘structural’’anti-aswellasthoseassociatedtostressresponses.Protectiveimmunityarisesagainstanti-gensthatareactivelyreleasedbygrowingbacillianditislikelythatthe‘‘focus’’oftheimmunolo-gicalresponseonlyongrowingbacilliallowsnonactivebacillitoremain‘‘invisible’’tothespeciřcimmunityofthehost.RUTIismadewithbacilligrownunderthestressfulconditionsofstarvation,lowandlowpH,conditionsachievedgraduallybyculturingonsolidmedia.Progresstostressfulconditions(e.g.,low)hasbeenshownto‘‘prepare’’thebacilliforevenmorestressfulconditions(e.g.,strictanaerobiosis),whileprogressivestarvationisknowntoleadtostationarygrowthinoldculturesinwhichslowmetabolismmakesbacillimoreresistanttostress,asinthechronicphaseofthemurineinfection.Insummary,bacilliusedtomakeRUTIaresubjectedtoconditionsthatareprobablyfoundinthegranulomaofhostswithactiveimmunity:alowinřbroticstructureofthegranulomaandthelowpHandstarvingconditionsinsidethephagolysosomeoftheacti-vatedmacrophage.ThechoiceoffragmentationofthebacillitomakeRUTIwastoallowoptimalpresentationofcellwallantigens,aswasthechoiceofcompositionintoliposomes.Theaveragediameterofthesefragments,whichis0.1m,allowstheantigensinthecellwalltobewellpresentedandthusprovideaneasierrecognitionoflatentbacilli.Moreover,itiswellknownthatthecellwallofM.tuberculosishasadjuvantpropertiesthatensuretheinductionofanimmunologicalresponsewithoutfurtheradjuvantmeasures.Inaddition,theinductionofapolyantigenicresponseispostulatedtohelprecog-nizelatentbacilli.AnotherrelevantfeatureofRUTIisthe‘‘detox-iřcation’’ofthecellfragmentsobtained.Inpreviousexperiments,ourřndingssupportedthehypothesisthatthecharacteristicINingranulomasM.tuberculosiswasinducednonspeciřcallybyendotoxin-likemolecules,essentiallyglycolipidslocatedintheoutermostlayerofthecellwallinsteadofbeinginducedbyaspeciřcimmunologi-calresponsesuchasdelayedtypehypersensitivityOurhypothesisarosefrompreviousworkbythegroupofG.A.W.Rook,suggestingthatINinducedbythe‘‘Kochphenomenon’’resemblesalocal‘‘Schwartzmanreaction’’.Thisnecrosisisnotusuallyseeninmurineexperimentalmodels,maybeduetotherelative‘‘tolerance’’ofthesesmallhoststoM.tuberculosisantigens.Fortheresponsetoinfectioninmicetobehavelikeother,larger,mammalsthatspontaneouslydevelopthisnecrosis,micewouldrequireahigheramountofendotoxin-likemolecules.Therefore,toavoidthepotentialriskofdevelopingaKochphenomenoncausedbyatherapeuticvaccineagainstLTBIwedecidedtoremoveendotoxin-likemoleculesfromthesurfaceofthefragmentsusingTritonadetergentthatminimizesthedenatura-tionofantigenicproteins.Finally,wealsoremovedthelipidicphasetoenhancethepresentationofthose‘‘hidden’’antigensbytheoutercoatandtoincreasetherangeoftheimmunologicalresponse.TheprotectionmechanismsofRUTIByusingawesternblottechnique,weobservedthattheinoculationof3dosesofRUTI(of185geach)2weeksapart,afterchemotherapy,inducedastrongpolyantigenicresponse,triggeringastrongTh1immunityagainstatleast13knownM.tuberculosisantigens,aswellasaTh3response.Inaddition,studiesonthecellularimmunityinducedby2dosesofRUTI(245g3weeksapart)andcomparedwiththeinoculationofBCGinmicepreviouslytreatedwithchemotherapy,revealedthatbothinoculationofBCGorRUTIinduceda10-foldincreaseintheCD4+populationinducedbyPPDstimulation.Interestingly,RUTIalsoenhancedtentimestheARTICLEINPRESS RUTI:treatmentoflatenttuberculosisinfection279 CD8IFN-+population(Fig.3),whereasthisincreasewasnotdetectedwithBCGwhichinduced

8 noprotection,adifferencealreadyfoundbyot
noprotection,adifferencealreadyfoundbyotherauthors.ThisdifferencemayexplaintheimprovedeffectivenessofRUTI,sincetheCD8IFN-populationiscrucialforthecontroloflatentbacilli.Additionally,antigenicdifferencesbe-tweenBCGandM.tuberculosisalsohelptoexplaindifferencesinprotection,whichwasanotherimportantfactorforthedesignofRUTI.AlthoughtheexactmechanismbywhichRUTIincreasestheCD8IFN+populationinthelungsremainstobeelucidated,itsdeliveryasliposomesappearscrucial;alternatively,theinductionofapoptosisinmacrophagesthatphagocyteRUTI,mayalsofavoraCD8response.Weweresurprisedtořndthatpolyclonalanti-bodiesobtainedafterRUTIadministrationprovidedprotectionagainstM.tuberculosisreactivationafterchemotherapyinSCIDmice.Intheseexperiments,weallowedaerosolinfectiontoprogressfor3weeks,atwhichpointchemotherapywithINHandrifampicinwasadministeredfor5weeks.Afterchemotherapy,twoexperimentalgroupsweredeřnedbytheinoculationofserumobtainedfrominfectedimmunocompetentmicetreatedwithchemotherapyandRUTI(serum-therapygroup),ortheinoculationofserumfromnoninfecteduntreatedimmunocompetentmice(controlgroup).After2weeks,necropsiesrevealedasigniřcantdifferencebetweenbothgroups:theserum-therapygroupreŖectedaweakerbacillaryreactivation(Fig.4),100timeslessthanthecontrol,andadecreasedgranulomatousinřltrationinthelungs(3timeslessthanthecontrol).Infact,micefromtheserum-therapygroupdevelopedfewerabscesses.ThesedatasupportthenotionthatIgGantibodiesarealsoabletocontroldisseminationoftheinfectioninthepulmonaryparenchyma,aswasearliersuggestedfromexperi-mentsofinfectionofbacillipre-coatedwithspeciřcIgG,orbyinducingactiveIgAproduc-Interestingly,ithasbeenrecentlyshownthatinoculationofnonspeciřcIgGalsoincreasestheeffectivenessofchemotherapywhenadminis-teredtherapeutically.Furthermore,astrongresponseofspeciřcantibodiesagainstlipoarabino-andAg85werealsoassociatedwithbetterprognosisinpatientswithactiveTB,furthersupportingthecurrentviewthatmereproductionofIFN-isinsufřcientforinductionofprotectionagainstTB,asreŖectedintheparadoxicalprotec-tionachievedbyCynomolgusmacaccawithBCGvaccination,whichcouldnotbelinkedtotheproductionofIFN-Acombinedcell/antibody-mediatedimmunityandcombinedTh1/Th2/Th3immunityisthoughttoprovidetheprotectionelicitedbyRUTI.Thistypeofresponseiscoherentwiththeconceptofglobalimmunityagainstlatentbacilli,withanaggressiveTh1cellularresponseprobablymediatedbyCD8+Tcells;Th2andTh3responsesthatwalloffthegranuloma79,80,110;andanantibodyTh1/Th2ARTICLEINPRESS 0810 123456 HS3 Figure4ControlofCFUsafterserum-therapyinthelungofSCIDmice.Afterinfection,miceweretreatedwithINH+RIFfromweek38(ingray),andtreatedwith4intraperitoneal(i.p.)inoculationsofserum(hyperim-muneserum,HS)frommicetreatedwithashortperiodchemotherapyandRUTI(inwhite)orfromnormalnoninfectedmice(inblack).Theresultsaregivenasmeanvalueswithstandarddeviationsobtainedfrom4miceforeachtimepoint(apartfromweek19,where10miceweretreatedwithimmuneserumand11miceweregivennormalserum).Differenceswithcontrolweresigniřcantwhenmarkedwith*for05. 3 cells 048406080100120 61417196141719 Figure3EvolutionofTCD4+andTCD8+IFN+cellsfromlungsinmicetreatedwithchemotherapyfromweeks6to14,andtreatedwith2subcutaneous(s.c.)inoculationsofRUTI(245g)atweeks14and17;1s.c.inoculationofBCG(10e6CFUs)atweek14or1s.c.ofemptyliposomeswhite,grayandblacksymbols,respectively.Theresultsaregivenasmeanvalueswithstandarddeviationsobtainedfrom4miceforeachtimepoint.Differenceswithcontrolweresigniřcantwhenmarkedwith*for05. P.-J.Cardona productionthatneutralizestheextracellularbacillithatescapethrou

9 ghthealveolarspaces.Theseconclusionsdono
ghthealveolarspaces.TheseconclusionsdonotagreewiththemostcommonhypothesissuggestingthatimmunityM.tuberculosisinfectionmustbeinducedbyaTh1response,whereasTh2responsesaredetrimental.ThishypothesisisbasedontheTh1/Th2responsetriggeredbyTBpatientscomparedwiththeresponsesseeninpatientswithLTBI,whoarethoughttoavoidreactivationoftheirlatentinfection.Inthisregard,sinceTh2responseplaysarelevantroleintheimmunopathologyofTBlesionsincontrollingandreclosingalargelesion(i.e.acavern)bybuildingastrongřbroticmantlearoundit,acompleteabsenceofanyTh2responsewouldbesurprising.Inanycase,sincemostoftheseřndingsareincidental,weconsiderthatthehypothesissuffersfroma‘‘chickenandegg’’problem:arethesepatientssufferingTBbecauseofTh2polarization,ordotheydevelopaTh2/Th3responsetofacethedisease?AnotherinterestingpointisthelackofefřcacyofBCGvaccinationinnewbornbabiesintheThirdWorldduetothelackofastrongTh1response.Thisseemstobecausedbytheinuterosensitizationtořlariae,whichresultsinaTh2bias,leadingtoaTh1/Th2responseinsteadofastrongerTh1responseasobservedagainstPPD-drivenIFN-production.Ontheotherhand,theimmunologi-calenvironmentatthefetomaternalinterfaceisTh2dominatedinordertoprotectthefetoplacen-talunit,andatthisagetheinductionofaTh2immunologicalresponseisgeneralized.Further-more,experimentalmicemodelsdonotsupportthishypothesis:nodifferenceineitherprotectionorintheprogressionofM.tuberculosisinfectionhasbeenfoundaftervaccinationwithdifferentdosesofBCGthatinducedeitherpureTh1ormixedTh1/Th2responses.Evenmoreso,nodifferencesintheevolutionofM.tuberculosisinfectioncouldbedemonstratedinIL-4,IL-4plusIL-13KO,IL-10KOmicecomparedwithwild-typemice.Finally,long-termchronicmurineTBinthelungswasnotinŖuencedbytheabsenceofBlymphocytesorIL-4.TheroleoftheTh3responsetriggeredbyRUTImustbeclariřed.ItispossiblethatTh3playsanimportantrolebeyondtheřnalřbrosisofthegranulomasthroughtheactionofTGF-fact,immunotherapywithDNAencodingforstressproteinslikehsp65hasproveneffectiveagainsttuberculosisinfectionwhencombinedwithche-motherapy.ItiswidelyacceptedthattheseproteinsinduceregulatoryTcellsthatcansuppressautoimmunityinducedbyTh1orTh2responses,thuscounterbalancingtheimmuneresponse.thisregard,theseproteinsmayhelpto‘‘synchro-nize’’theimmuneresponse(i.e.activatingtheinfectedmachrophagesorwallingoffthegranulo-mas)ratherthanjustinducingastrongTh1responseorreducingtheTh2response,similartothepostulatedmechanismofMycobacteriumimmunotherapy.Whyispreviousshort-termchemotherapystillnecessarybeforeLTBItherapeuticvaccination?Atthispoint,itisveryimportanttostudyLTBIinhumanpatients,becausewecurrentlyignorewhichbacillarypopulationthelesionsharbor.Aremostofthesebacilliactivelygrowingoraretheylatent?Andwhichistherelativeproportionofthesepopulations?Thepresenceofheterogeneouspopu-lationsinalesionmayexplainwhyashortperiodofchemotherapy(e.g.4weeks),whichcould‘‘har-monize’’or‘‘standardize’’thebacillarypopulationinallpatientstowardalatentpopulation,isadvantageousfortherapy.Furthermore,thisshortperiodofchemotherapyalsoallowsadecreaseintheimmunologicalconstantboostagainstgrowingbacilliandremovesFM(Fig.5)andthusthesourceoflocalimmuno-suppression.Moreover,chemotherapyinpatientswithLTBIhasalsobeenassociatedwiththeincreaseofTh1speciřceffectorcellsinperipheralbloodwithintheřrst4weeksofLTBIconventionalchemotherapy(9monthsofINH).ThisriseinTh1effectorcellsarisesfromantigenpresentationandthereleaseofeffectorTcellsfromtheinfectedThesedataalsosupportthehypothesisthatlocalchangesinthegranulomafavourrecogni-tionoflatentbacilli.Chemotherapyalsoi

10 ncreasesreabsorptionofthelesion,therefor
ncreasesreabsorptionofthelesion,thereforereducingtheinŖammatoryresponseinthelungs,reducingthechancethatthetherapeuticvaccinewillinduceaKochphenomenon,becausetheinfectedsitewillnolongerbea‘‘preparedsite’’.Inconclusion,theneedforchemotherapyin-dicatesthatoncelatentbacilliareinduced,vaccinationisuselessbecausethesebacilliremaininvisibletotheimmunesystem.Moreover,evenifthehost’simmunityisalreadypreparedtocontrolinfection,itcannotpreventthereactivationofthesecellsinasitewhichisprivilegedforthebacilli(suchasthepulmonaryapex),thusbeingunabletopreventtheinductionofpulmonaryTB.Therefore,itispossiblethatchemotherapyallowsthepresentationoflatentbacillitothehostimmunityby‘‘rediscovering’’theirpresence.ARTICLEINPRESS RUTI:treatmentoflatenttuberculosisinfection281 HowhastheeffectivenessofRUTIbeenTheresearchofnewdrugsandvaccinesagainstTBisespeciallydifřcultbecausenodirectcorrelationexistsbetweentheresultsobtainedincommonlyusedexperimentalmodelsandwhatreallyhappensinhumans.Forinstance,testingnewprophylacticvaccinesrequiresanefřcacyatleastsimilartothatobtainedwithBCGvaccination,whichreducesbacterialconcentrationinmouselungsby1logat3weeksafterinfectioncomparedtononvacci-natedanimalsandallowsinfectedguineapigstosurviveforupto30weeksinsteadofthe14weeksusuallyseeninnonvaccinatedanimals.Overall,prophylacticvaccinesdonotprovideanyprophy-laxisbutimprovecontroloftheinfection,althoughtheydonotpreventdeathofinfectedanimalsasaresultofTBdevelopment.Thelackofclearcorrelationbetweeneffectivenessofvaccinesinanimalmodelsandinhumansmeansthatthevalidationofnewtherapiesinexperimentalmodelsmustbeinterpretedwithcaution.Forexample,inLTBItherapy,thegoldstandardtreatmentensuresa90%efřcacyagainstLTBIreactivationinhumansbutatmostitonlyachievesa10%efřcacyinAnotherstrikingandrelevantquestionalreadydiscussedaboverelatestothetypeofexperimentalmodelthatneedstobeusedfortestingtheefřcacyofanewtherapeuticvaccine,withtheaimofmimickingwhathappensinhumans.TheCornellmodeldoesnotappearveryappropriatebecauseitisnotclearifpersistentorlatentbacilliarepresent,andno‘‘complete’’granulomasandgrowingbacilliaredetectedattheonsetoftreatment.ThusitdoesnotreŖecttheconditionsofhumanLTBI.Instead,wechosechronicinfectioninmiceasastartingpoint.Moreover,the‘‘toler-ance’’shownbymicetoM.tuberculosisinfectionARTICLEINPRESS Figure5Effectsof8-weekchemotherapyonthestructureofgranulomas.NotethelackofFMintheperipheryoftreatedmice(B)comparedwithmicebeforestartingchemotherapy(A),atweek8post-aerosolinfection.Slideswerestainedwithhematoxylineosin.Originalampliřcationis400x(datanotpublished). 039171922 12 H+RRUTI Figure6BactericidalactivityofRUTIinthelungofC57BL/6mice.Afterinfection,miceweretreatedwithINH+RIFfromweek917(ingray),andwith3subcuta-neous(s.c.)inoculationsofRUTI(185gperinoculation)atweeks17,19and21(inwhite)oremptyliposomes(inblack).Theresultsaregivenasmeanvalueswithstandarddeviationsobtainedfrom4to6miceforeachtimepoint.Differenceswithcontrolweresigniřcantwhenmarkedwith*for05(modiředfrom P.-J.Cardona mayunderestimatenewtreatments,asthehostdoesnotdisplayastronginŖammatoryresponseagainstthebacilli,suggestingthatevaluatingthereductionofbacillarybulkanditscontrolafterchemotherapyshouldbetheřnaltargetinsteadofsterilization.ItisrelevantthatinitialexperimentsshowedthatRUTIsigniřcantlyreducedthebacil-larybulk(Fig.6andmaintainedthiscontrolforalongtime,atleastforupto11weeksafterthelastRUTIinoculation.Concerningthe‘‘tolerance’’phenomenondis-cussedabove,wedecidedtorunnewexperimentswithguineapigsinsteadofmice.Therationaleforth

11 isdecisionwasbasedonthehypothesisthatthe
isdecisionwasbasedonthehypothesisthatthebiggerthehost,thestrongertheinŖammatoryresponseagainstM.tuberculosis,becausemoreparenchymacanbedestroyedinlargeranimalstostopbacillarygrowth(assumingthatthesizeofimmunecellsissimilarinallmammals).Fig.7exempliřesthisideabycomparingthevolumeofasmallhumanTBcavernwiththetotalvolumesofdifferentmammals,andshowsthatamousewillneverdevelopsuchalesion,asitrepresentsitstotalvolume.Wethusstartedworkingwiththeguineapigusingamore‘‘humanized’’,shortchemotherapyregimen(4weekswithINHandrifampicin),aswellasamorevirulentM.tubercu-strain(H37RvPasteur).Histopathologicalanalysis4weeksafterinfectionshowedthatguineapigsdevelopedastructuredanddeřnedgranulo-ma,withstrongoutermostřbrosisandIN;fewacidfast-bacilliweredetected,localizedonlyintheperipheryofthegranulomaandinsideFM(Fig.8Incontrast,atthissametimepoint,miceonlyARTICLEINPRESS Figure7Comparisonofdifferenthostsaccordingtotheirweight.ThevolumeofaTBcavernweighing25gisshownin Figure8Comparisonoflesionsinguineapigs(AD)andmice(EH)atweek4post-aerosolinfection.PicturesA,B,EandFshowthestructureofthegranulomastainedwithhaematoxylineosin.Notethehighlyorganizedguineapiggranuloma,thepresenceofacentralnecrotictissuesurroundedbyamantleofřbrotictissueandanoutermostlayerdominatedbylymphocytes.Incontrast,thegranulomaofmiceisjustamixtureofPMNs,macrophagesandsomelymphocyteswithnoapparentorganization.PicturesC,D,GandHshowtheresultofZiehlNeelsenstaininginthesegranulomas.Acid-fastbacilliarescarceinsideFMincludedintheřbroticmantle(C),andinsidethenecroticcenter(D);onthecontrary,acidfastbacillicanbeeasilyseeninmicegranuloma(GandH). RUTI:treatmentoflatenttuberculosisinfection283 developedatinypre-granulomafullofacid-fastbacilli,withnoouterřbroticcontent,thusleavingmoretimeforfurtherpulmonarydissemination.Infact,bacillarycountswerehigherinmicethaninguineapigs(1logmore),whichagreewithpreviousreports.Inthiscontext,administrationof3dosesofRUTI(180g)atweeks8,10and14aftershorterchemotherapythaninmiceexperi-ments(betweenweeks4and8)signiřcantlydecreasedthebacillaryloadatweek16(atthetimeofnecropsy)evenwhenbacillarybulkincontrolguineapigsdidnotincrease(Fig.9).Incontrast,bacillarycountsincontrolmiceincreasedby2log10whenthesameM.tuberculosiswasused.NotethatcontrolofthebacillarycountinguineapigsrequiresatremendousinŖammatoryeffortthatřnallykillsthehostfasterthanitkillsmice.Therefore,wecannotassumethatmicearemoreresistanttoM.tuberculosisinfectionthanguineapigs,butinstead,weassumethatmicearemore‘‘tolerant’’tothesebacilli,allowinganabundanceofbacilliintheirlungsbytriggeringaweakerinŖammatoryresponse.ThistoleranceallowsmicetosurvivelongertoM.tuberculosisinfection,butintheendthebacillialsokillthehostduetoconstantdisseminationthroughouttheirHowever,itisobviousthattheevolutionofinfectionisnotequivalentinguineapigsandinhumanhostsbecausethelattercontrolbacillaryprogressionbetterbywallingthemoffinthegranulomas.Theformationofgranulomasinhumanswillalsoallowadministrationofweakerchemotherapy(e.g.INHalone)ratherthanthecombinationofINHandrifampicinrequiredforcontrollinginfectioninguineapigs.ARTICLEINPRESS 08101416 12 H+RRUTI 4 Figure9BactericidalactivityofRUTIinthelungofguineapigs.Afterinfection,animalsweretreatedwithINH+RIFfromweek48(ingray),andwith3subcuta-neous(s.c.)inoculationsofRUTI(185gperinoculation)atweeks8,10and14(inwhite)oremptyliposomes(inblack).Theresultsaregivenasmeanvalueswithstandarddeviationsobtainedfrom8animalsforeachtimepoint.Differenceswithcontrolweresigniřcantwhenmarkedwith*for05. Table1PropertiesoftheL

12 TBI.A.ThenatureoflatentbacilliMycobacter
TBI.A.ThenatureoflatentbacilliMycobacteriumtuberculosishasainnateslowgrowth-rate.Oncesubmittedtostress,bacillislowmetabolismtonear-zeroactivity,inwhichsituationthebacillicanresiststressfulenvironments.Arelevantcapacitytosurviveinnecrotictissue.Bacillicangrowattheperipheryofthegranulomasoncethestresshasresumed.B.ThehostimmunologicalresponseThehostdevelopactiveimmunitylargelyagainstgrowingbacilli.CD8Tcellsmayplayarelevantroleinthecontroloflatentbacilli.C.TheroleoffoamymacrophagesAreaconsequenceoftheaccumulationofnecroticdebris,surfactantanddeadM.tuberculosiscellsHaveadecreasedabilityforpresentingantigens.SuppresseffectorTcellsthattrytoactivatethem.Harbourlatentbacilli.Representtheoutermostlayerofthegranuloma.Rendertheinductionofaspeciřcimmunologicalresponseagainstlatentbacilliuseless.KeyaspectsforthedestructionofM.tuberculosislatentbacilliRemovaloftheoutermostlayeroffoamymacrophagesthatexertlocalimmunosuppression.Inductionofapolyantigenicimmunologicalresponsenotonlyagainstantigensofactivelygrowingbacilli,butalsoagainststructuralantigens. P.-J.Cardona RegardingthepossibletoxicityofRUTI,inbothexperimentswithmiceandguineapigs,onlyalocaltransientinŖammatoryresponseatthesiteofinoculationwasfound.Nosystemictoxicity(weightlossorincreasedgranulomatousresponse)weredetected.ThefutureBasedoncurrentdata,newexperimentsarebeingcarriedouttogainfurtherunderstandingofallthemechanismsinvolvedintheprotectivebeneřtsofRUTI.Theseexperimentsincludemore‘‘huma-nized’’modelswithlargermammalianhosts.Inthisregard,wehavestartedsomeřeldexperimentswithnaturallyinfectedgoatsandexperimentalmodelssuchasmini-pigs.Ouraimistousehostswithavolumemorecomparabletohumansthanmodelsusedhitherto,todeterminetheprotectiveeffectsofRUTIandalsotoconřrmlackoftoxicity,becauselargerhostsaremorelikelytodevelopgreaterinŖammatoryresponsesandtheKochphenomenon.Inconclusion,weexpecttoachieveabetterchemotherapeutictreatmentofLTBIpatients,byadministeringINHforashortperiodoftime,i.e.4weeks,withonly2dosesofRUTI;thisimprovedregimenisalsoexpectedtoamelioratetreatmentcompliance.(Table1AknowledgementsI’mindebtwithRajkoReljic,JurajIvanyiandIreneGarcia-Gabayfortheircontributionstothepaper.1.WayneLG,SramekHA.MetronidazoleisbactericidaltodormantcellsofMycobacteriumtuberculosisAntimicrobAgentsChemother2.WayneLG,SohaskeyCD.Nonreplicatingpersistenceofmycobacteriumtuberculosis.AnnuRevMicrobiol3.WayneLG,LinKY.GlyoxylatemetabolismandadaptationofMycobacteriumtuberculosistosurvivalunderanaerobicInfectImmun4.MochizukiM.Kineticsofoxygenandcarbondioxidereactions.CrystalRG,WestJB,etal.,editors.Thelung:scienticfoundations,vol.5.3.2.4.NewYork:RavenPressLtd;1991.p.12415.CooperAM,DaltonDK,StewartTA,GrifřnJP,RussellDG,OrmeIM.Disseminatedtuberculosisininterferongammagene-disruptedmice.JExpMed6.GilO,GuiradoE,GordilloS,etal.IntragranulomatousnecrosisinlungsofmiceinfectedbyaerosolwithMycobacteriumtuberculosisisrelatedtobacterialloadratherthantoanyonecytokineorTcelltype.MicrInfectAvailableonline17January2006.7.McCuneRM,FeldmannFM,LambertHP,McDermottW.MicrobialpersistenceIThecapacityoftuberclebacillitosurvivesterilizationinmousetissues.JExpMed8.AmericanThoracicSocietyandtheCentersforDiseaseControlandPrevention.Targetedtuberculintestingandtreatmentoflatenttuberculosisinfection.AmJRespirCritCareMed9.LucasSB.Histopathology.DaviesPDO,editor.Clinicaltuber-,vol.8.London:Chapman&Hall;1998.p.11327.10.GomezJE,McKinneyJD,editors.M.tuberculosispersis-tencelatencyanddrugtoleranceTuberculosis(Edinburgh)11.OrmeIM.ThelatenttuberculosisbacillusI’l

13 lletyouknowifIevermeetone.IntJTubercLung
lletyouknowifIevermeetone.IntJTubercLungDis12.WallisRS,PatilS,CheonSH,etal.DrugtoleranceinMycobacteriumtuberculosisAntimicrobAgentsChe-13.ShleevaMO,BagramyanK,TelkovMV,etal.Formationandresuscitationof‘‘non-culturable’’cellsofRhodococcusrhodochrousandMycobacteriumtuberculosisinprolongedstationaryphase.Microbiology14.KellDB,YoungM.Bacterialdormancyandculturability:theroleofautocrinegrowthfactors.CurrOpinMicrobiol15.WallaceJG.Theheatresistanceoftuberclebacilliinthelungsofinfectedmice.AmRevRespirDis16.Munųoz-EliasEJ,TimmJ,BothaT,ChanWT,GomezJE,McKinneyJD.ReplicationdynamicsofMycobacteriuminchronicallyinfectedmice.InfectImmun17.LazarevicV,NoltD,FlynnJL.Long-termcontrolofMycobacteriumtuberculosisinfectionismediatedbydynamicimmuneresponses.JImmunol18.ThoenCO.Tuberculosisinwildanddomesticmammals.BloomBR,editor.Tuberculosispathogenesisprotectictionandcontrol,vol.11.Washington,DC:ASMPress;1994.p.15719.RookGA,AttiyahR.CytokinesandtheKochphenomenon.Tubercle20.CardonaPJ,LlatjosR,GordilloS,etal.Towardsa‘human-like’modeloftuberculosis:intranasalinoculationofLPSinducesintragranulomatouslungnecrosisinmiceinfectedaerogenicallywithMycobacteriumtuberculosisJImmunol21.RyllR,KumazawaY,YanoI.Immunologicalpropertiesoftrehalosedimycolatecordfactorandothermycolicacid-containingglycolipidsareview.MicrobiolImmunol22.IndrigoJ,HunterJr.RL,ActorJK.Cordfactortrehalose66’-dimycolateTDMmediatestrafřckingeventsduringmycobacterialinfectionofmurinemacrophages.23.FlynnJL,ChanJ.Immunologyoftuberculosis.AnnuRev24.OrmeIM,editor.ThemouseasausefulmodelofTuberculosis(Edinburgh)25.NorthRJ,JungYJ.Immunitytotuberculosis.AnnuRev26.CardonaPJ,LlatjosR,GordilloS,etal.EvolutionofgranulomasinlungsofmiceinfectedaerogenicallywithMycobacteriumtuberculosisScandJImmunolARTICLEINPRESS RUTI:treatmentoflatenttuberculosisinfection285 27.CardonaPJ,GordilloS,DiazJ,etal.WidespreadbronchogenicdisseminationmakesDBA/2micemoresusceptiblethanC57BL/6micetoexperimentalaerosolinfectionwithMycobacteriumtuberculosisInfectImmun28.GreenGM,BurlingtonVT.Alveolobronchiolartransportmechanisms.ArchInternMed29.KorfJ,StoltzA,VerschoorJ,DeBaetselierP,GrootenJ.Mycobacteriumtuberculosiscellwallcomponentmycolicacidelicitspathogen-associatedhostinnateimmuneresponses.EurJImmunol30.ErikssonU,EgermannU,BihlMP,etal.HumanbronchialepitheliumcontrolsTH2responsesbyTH1-induced,nitricoxide-mediatedSTAT5dephosphorylation:implicationsforthepathogenesisofasthma.JImmunol31.MillsCD.Molecularbasisof"suppressor"macrophagesArgininemetabolismviathenitricoxidesynthetasepath-way.JImmunol32.BingisserRM,TilbrookPA,HoltPG,KeesUR.Macrophage-derivednitricoxideregulatesTcellactivationviareversibledisruptionoftheJak3/STAT5signalingpathway.JImmunol33.NabeshimaS,NomotoM,MatsuzakiG,etal.T-CellhyporesponsivenessinducedbyactivatedmacrophagesthroughnitricoxideproductioninmiceinfectedwithMycobacteriumtuberculosisInfectImmun34.ChanED,ChanJ,SchlugerNW.Whatistheroleofnitricoxideinmurineandhumanhostdefenseagainsttubercu-losis?Currentknowledge.AmJRespirCellMolBiol35.FlynnJL,ScangaCA,TanakaKE,ChanJ.Effectsofaminoguanidineonlatentmurinetuberculosis.JImmunol36.LamasS,MichelT,BrennerBM,MarsdenPA.Nitricoxidesynthesisinendothelialcells:evidenceforapathwayinduciblebyTNF-alpha.AmJPhysiol37.NusslerAK,DiSilvioM,BilliarTR,etal.Stimulationofthenitricoxidesynthasepathwayinhumanhepatocytesbycytokinesandendotoxin.JExpMed38.ChuSC,Marks-KonczalikJ,WuHP,BanksTC,MossJ.Analysisofthecytokine-stimulatedhumaninduciblenitricoxidesynthase(iNOS)gene:characterizationofdifferencesbetweenhuman

14 andmouseiNOSpromoters.BiophysResCommun39
andmouseiNOSpromoters.BiophysResCommun39.KolbJP,Paul-EugeneN,DamaisC,YamaokaK,DrapierJC,DugasB.Interleukin-4stimulatescGMPproductionbyIFN-gamma-activatedhumanmonocytes.Involvementofthenitricoxidesynthasepathway.JBiolChem40.GuoFH,UetaniK,HaqueSJ,etal.Interferongammaandinterleukin4stimulateprolongedexpressionofinduciblenitricoxidesynthaseinhumanairwayepitheliumthroughsynthesisofsolublemediators.JClinInvest41.EiglerA,MoellerJ,EndresS.ExogenousandendogenousnitricoxideattenuatestumornecrosisfactorsynthesisinthemurinemacrophagecelllineRAW264.7.JImmunol42.ErikssonU,EgermannU,BihlMP,etal.HumanbronchialepitheliumcontrolsTH2responsesbyTH1-induced,nitricoxide-mediatedSTAT5dephosphorylation:implicationsforthepathogenesisofasthma.JImmunol43.CooperAM,AdamsLB,DaltonDK,AppelbergR,EhlersS.IFN-gammaandNOinmycobacterialdisease:newjobsforoldhands.TrendsMicrobiol44.Hernandez-PandoR,SchonT,OrozcoEH,SerařnJ,Estrada-GarciaI.Expressionofinduciblenitricoxidesynthaseandnitrotyrosineduringtheevolutionofexperimentalpulmon-arytuberculosis.ExpToxicolPathol45.YuK,MitchellC,XingY,MagliozzoRS,BloomBR,ChanJ.Toxicityofnitrogenoxidesandrelatedoxidantsonmycobacteria:M.tuberculosisisresistanttoperoxynitriteTuberLungDis46.RobinsonVK,SatoE,NelsonDK,CamhiSL,RobbinsRA,HoytJC.Peroxynitriteinhibitsinducible(type2)nitricoxidesynthaseinmurinelungepithelialcellsinvitro.FreeRadicBiolMed47.MoriwakiH,KumeN,KataokaH,etal.Expressionoflectin-likeoxidizedlowdensitylipoproteinreceptor-1inhumanandmurinemacrophages:upregulatedexpressionbyTNF-FEBSLett48.SmirnovaIV,KajsturaM,SawamuraT,GoligorskyMS.AsymmetricdimethylarginineupregulatesLOX-1inacti-vatedmacrophages:roleinfoamcellformation.JPhysiolHeartCircPhysiol49.KruthHS,HuangW,IshiiI,ZhangWY.Macrophagefoamcellformationwithnativelowdensitylipoprotein.JBiolChem50.LiAC,GlassCK.Themacrophagefoamcellasatargetfortherapeuticintervention.NatMed51.BurleighME,BabaevVR,OatesJA,etal.Cyclooxygenase-2promotesearlyatheroscleroticlesionformationinLDLreceptor-deřcientmice.52.WrightJR.Immunomodulatoryfunctionsofsurfactant.PhysiolRev53.LianX,YanC,YangL,XuY,DuH.LysosomalacidlipasedeřciencycausesrespiratoryinŖammationanddestructioninthelung.AmJPhysiolLungCellMolPhysiol54.KramerBW,JobeAH,IkegamiM.Exogenoussurfactantchangesthephenotypeofalveolarmacrophagesinmice.AmJPhysiolLungCellMolPhysiol55.ChanED,MorrisKR,BelisleJT,etal.Inductionofinduciblenitricoxidesynthase-NO*bylipoarabinomannanofbacteriumtuberculosisismediatedbyMEK1-ERK,MKK7-JNK,andNF-kappaBsignalingpathways.InfectImmun56.RhoadesER,OrmeIM.SusceptibilityofapanelofvirulentstrainsofMycobacteriumtuberculosistoreactivenitrogenintermediates.InfectImmun57.BrugmannWB,FirmaniMA.LowconcentrationsofnitricoxideexertahormeticeffectonMycobacteriumtubercu-invitro.JClinMicrobiol58.CicconeR,MarianiF,CavoneA,etal.InhibitoryeffectofNO-releasingCiproŖoxacin(NCX976)onMycobacteriumtuberculosissurvival.AntimicrobAgentsChemother59.VoskuilMI,SchnappingerD,ViscontiKC,etal.InhibitionofrespirationbynitricoxideinducesaMycobacteriumdormancyprogram.JExpMed60.SchnappingerD,EhrtS,VoskuilMI,etal.TranscriptionalAdaptationofMycobacteriumtuberculosiswithinMacro-phages:InsightsintothePhagosomalenvironment.JExp61.OhnoH,ZhuG,MohanVP,etal.TheeffectsofreactivenitrogenintermediatesongeneexpressioninMycobacter-iumtuberculosisCellMicrobiol62.DarwinKH,EhrtS,Gutierrez-RamosJC,WeichN,NathanCF.TheproteasomeofMycobacteriumtuberculosisARTICLEINPRESS P.-J.Cardona requiredforresistancetonitricoxide.63.Hernandez-PandoR,OrozcoH,ArriagaK,SampieriA,Larriva-SahdJ,Madrid

15 -MarinaV.Analysisofthelocalkineticsandlo
-MarinaV.Analysisofthelocalkineticsandlocalizationofinterleukin-1alphatumournecrosisfactor-alphaandtransforminggrowthfactor-betaduringthecourseofexperimentalpulmonarytuberculosis.64.OrdwayD,Henao-TamayoM,OrmeIM,Gonzalez-JuarreroM.FoamymacrophageswithinlunggranulomasofmiceinfectedwithMycobacteriumtuberculosisexpressmole-culescharacteristicofdendriticcellsandantiapoptoticmarkersoftheTNFreceptor-associatedfactorfamily.JImmunol65.CardonaPJ,JulianE,VallesX,etal.ProductionofantibodiesagainstglycolipidsfromtheMycobacteriumcellwallinaerosolmurinemodelsoftuberculosis.ScandJImmunol66.ShalitI,Horev-AzariaL,FabianI,etal.ImmunomodulatoryandprotectiveeffectsofmoxiŖoxacinagainstCandidaalbicans-inducedbronchopneumoniainmiceinjectedwithcyclophosphamide.AntimicrobAgentsChemother67.WerberS,ShalitI,FabianI,SteuerG,WeissT,BlauH.MoxiŖoxacininhibitscytokine-inducedMAPkinaseandNF-kappaBactivationaswellasnitricoxidesynthesisinahumanrespiratoryepithelialcellline.JAntimicrobChe-68.DunnPL,NorthRJ.VirulencerankingofsomeMycobacter-iumtuberculosisandMycobacteriumbovisstrainsaccord-ingtotheirabilitytomultiplyinthelungsinducelungpathologyandcausemortalityinmice.InfectImmun69.GrodeL,SeilerP,BaumannS,etal.IncreasedvaccineefřcacyagainsttuberculosisofrecombinantMycobacter-iumbovisbacilleCalmette-GuerinmutantsthatsecreteJClinInvest70.RamachandraL,NossE,BoomWH,HardingCV.ProcessingMycobacteriumtuberculosisantigen85Binvolvesin-traphagosomalformationofpeptide-majorhistocompat-ibilitycomplexIIcomplexesandisinhibitedbylivebacillithatdecreasephagosomematuration.JExpMed71.RamachandraL,SmialekJL,ShankSS,ConveryM,BoomWH,HardingCV.PhagosomalprocessingofMycobacteriumantigen85Bismodulatedindependentlyofmycobacterialviabilityandphagosomematuration.72.SchaibleUE,WinauF,SielingPA,etal.ApoptosisfacilitatesantigenpresentationtoTlymphocytesthroughMHC-IandCD1intuberculosis.NatMed73.TurnerOC,BasarabaRJ,OrmeIM.ImmunopathogenesisofpulmonarygranulomasintheguineapigafterinfectionMycobacteriumtuberculosisInfectImmun74.UlrichsT,KosmiadiGA,JorgS,etal.Differentialorganiza-tionofthelocalimmuneresponseinpatientswithactivecavitarytuberculosisorwithnonprogressivetuberculoma.JInfectDis75.FisherMA,PlikaytisBB,ShinnickTM.MicroarrayanalysisofMycobacteriumtuberculosistranscriptionalresponsetotheacidicconditionsfoundinphagosomes.JBacteriol76.ColstonMJ,CoxRA.Mycobacterialgrowthanddormancy.RatledgeC,DaleJ,editors.Mycobacteriamolecularbiologyandvirulence,vol.11.London:BlackwellScience;1999.p.19877.CardonaPJ,Ruiz-ManzanoJ.OnthenatureofMycobacter-iumtuberculosis-latentbacilli.EurRespirJ78.RookGA,Hernandez-PandoR,DhedaK,TengSeahG.IL-4intuberculosis:implicationsforvaccinedesign.Trends79.DhedaK,BoothH,HuggettJF,JohnsonMA,ZumlaA,RookGA.LungRemodelinginPulmonaryTuberculosis.JInfect80.AungH,ToossiZ,McKennaSM,etal.Expressionoftransforminggrowthfactor-betabutnottumornecrosisfactor-alfainterferon-gammaandinterleukin-4ingranulo-matouslunglesionsintuberculosis.TubercleLungDis81.FenhalsG,WongA,BezuidenhoutJ,vanHeldenP,BardinP,LukeyPT.Insituproductionofgammainterferon,inter-leukin-4,andtumornecrosisfactoralphamRNAinhumanlungtuberculousgranulomas.InfectImmun82.ComstockGW.Howmuchisoniazidisneededforpreventionoftuberculosisamongimmunocompetentadults?JTubercLungDis83.JindaniA,DoreCJ,MitchisonDA.Bactericidalandsterilizingactivitiesofantituberculosisdrugsduringtheřrst14days.AmJRespirCritCareMed84.MitchisonDA.Basicmechanismsofchemotherapy.85.GrossetJ.Bacteriologicbasisofshort-coursechemother-apyfortuberculosis.ClinChestMed86.OlsenAW,Brand

16 tL,AggerEM,vanPinxterenLA,AndersenP.Thei
tL,AggerEM,vanPinxterenLA,AndersenP.TheinŖuenceofremainingliveBCGorganismsinvaccinatedmiceonthemaintenanceofimmunitytotuberculosis.ScandJImmunol87.CardonaPJ,AmatI,GordilloS,etal.ImmunotherapywithMycobacteriumtuberculosiscellsincreasestheeffectivenessofchemotherapyagainstachronicalinfec-tioninamurinemodeloftuberculosis.Vaccine88.OrmeIM,AndersenP,BoomWH.TcellresponsetoMycobacteriumtuberculosisJInfectDis1993;167:148197.89.PetersAC,WimpennyJW,CoombsJP.OxygenprořlesinandintheagarbeneathcoloniesofBacilluscereusStaphylococcusalbusandEscherichiacoli.JGenMicrobiol90.RobinsonTP,WimpennyJW,EarnshawRG.pHgradientsthroughcoloniesofBacilluscereusandthesurroundingagar.JGenMicrobiol91.LesermanL.Liposomesasproteincarriersinimmunology.LiposomeRes92.LarsonCL,BellJF,ListRH,RibiE,WichtWC.SymposiumonrelationshipofstructureofmicroorganismstotheirimmunologicalpropertiesIIhost-reactivepropertiesofcellwallsandprotoplasmfrommycobacteria.BacteriolRev93.DannenbergJrAM.Rolesofcytotoxicdelayed-typehypersensitivityandmacrophage-activatingcell-mediatedimmunityinthepathogenesisoftuberculosis.94.MoreiraAL,TsenovaL,AmanMH,etal.MycobacterialantigensexacerbatediseasemanifestationsinMycobacter-iumtuberculosis-infectedmice.InfectImmun95.TaylorJL,OrdwayDJ,TroudtJ,Gonzalez-JuarreroM,BasarabaRJ,OrmeIM.FactorsassociatedwithsevereARTICLEINPRESS RUTI:treatmentoflatenttuberculosisinfection287 granulomatouspneumoniainMycobacteriumtuberculosisinfectedmicevaccinatedtherapeuticallywithhsp65DNA.InfectImmun96.BrennanPJ.Structure,function,andbiogenesisofthecellwallofMycobacteriumtuberculosis.Tuberculosis(Edin-97.GilO,GuiradoE,AmatI,etal.Thecontroloflatenttuberculosisinfectionafterashort-coursechemotherapyinducedbyRUTIisrelatedtotheincreaseofPPD-speciřcCD8+cellsinthelungsandthereductionofpulmonarydisseminationthroughapoliclonalhumoralresponseNewapproachestovaccinedevelopment,Berlin,September98.TurnerJ,RhoadesER,KeenM,BelisleJT,FrankAA,OrmeIM.EffectivepreexposuretuberculosisvaccinesfailtoprotectwhentheyaregiveninanimmunotherapeuticInfectImmun99.vanPinxterenLA,CassidyJP,SmedegaardBH,AggerEM,AndersenP.ControloflatentMycobacteriumtuberculosisinfectionisdependentonCD8Tcells.EurJImmunol100.AggerEM,AndersenP.AnovelTBvaccine;towardsastrategybasedonourunderstandingofBCGfailure.Vaccine101.SoruriA,SchweyerS,RadzunHJ,FayyaziA.Mycobacterialantigensinduceapoptosisinhumanpuriředproteinderivative-speciřcalphabetaTlymphocytesinaconcentration-dependentmanner.102.SchaibleUE,WinauF,SielingPA,FischerK,CollinsHL,HagensK,ModlinRL,BrinkmannV,KaufmannSH.ApoptosisfacilitatesantigenpresentationtoTlymphocytesthroughMHC-1andCDIintuberculosis.NatMed103.GuiradoE,AmatI,GilO,etal.Passiveserum-therapywithpolyclonalantibodiesagainstMycobacteriumtuberculosisprotectsagainstpost-chemotherapyrelapseoftuberculosisinfectioninSCIDmice.MicrobInfect.Availableonline27January2006.104.Glatman-FreedmanA.Advancesinantibody-mediatedimmunityagainstMycobacteriumtuberculosis:implica-tionsforanovelvaccinestrategy.FEMSImmunolMed105.WilliamsA,ReljicR,NaylorI,etal.PassiveprotectionwithimmunoglobulinAantibodiesagainsttuberculousearlyinfectionofthelungs.106.RoyE,StavropoulosE,BrennanJ,etal.Therapeuticefřcacyofhigh-doseintravenousimmunoglobulininMycobacteriumtuberculosisinfectioninmice.107.CostelloAM,KumarA,NarayanV,etal.Doesantibodytomycobacterialantigensincludinglipoarabinomannanlimitdisseminationinchildhoodtuberculosis?TransRSocTropMedHyg108.Sanchez-RodriguezC,Estrada-ChavezC,Garcia-VigilJ,etal.AnIgGantibodyresponsetotheantigen85complexisassociatedw

17 ithgoodoutcomeinMexicanTotonacaIndianswi
ithgoodoutcomeinMexicanTotonacaIndianswithpulmonarytuberculosis.IntJTubercLungDis109.LangermansJA,AndersenP,vanSoolingenD,etal.DivergenteffectofbacillusCalmette-GuerinBCGvaccina-tiononMycobacteriumtuberculosisinfectioninhighlyrelatedmacaquespecies:implicationsforprimatemodelsintuberculosisvaccineresearch.ProcNatlAcadSciUSA110.SimePJ,XingZ,GrahamFL,CsakyKG,GauldieJ.Adenovector-mediatedgenetransferofactivetransform-inggrowthfactor-beta1inducesprolongedsevereřbrosisinratlung.JClinInvest111.DemissieA,AbebeM,AseffaA,etal.HealthyindividualsthatcontrolalatentinfectionwithMycobacteriumexpresshighlevelsofTh1cytokinesandtheIL-4antagonistIL-4delta2.JImmunol112.MalhotraI,MungaiP,WamachiA,etal.Helminth-andBacillusCalmette-Guerin-inducedimmunityinchildrensensitizedinuterotořlariasisandschistosomiasis.JImmunol113.PrescottSL,MacaubasC,HoltBJ,etal.Transplacentalprimingofthehumanimmunesystemtoenvironmentalallergens:universalskewingofinitialTcellresponsestowardtheTh2cytokineprořle.JImmunol114.GruppoV,OrmeIM,editors.DoseofBCGdoesnotinuencetheefcientgenerationofprotectiveimmunityinmicechallengedwithMycobacteriumtuberculosisTuberculosis115.JungYJ,LaCourseR,RyanL,NorthRJ.Evidenceincon-sistentwithanegativeinŖuenceofThelper2cellsonprotectionaffordedbyadominantThelper1responseMycobacteriumtuberculosislunginfectioninmice.InfectImmun116.JungYJ,RyanL,LaCourseR,NorthRJ.Increasedinterleukin-10expressionisnotresponsibleforfailureofThelper1immunitytoresolveairborneMycobacteriuminfectioninmice.117.TurnerJ,FrankAA,BrooksJV,Gonzalez-JuarreroM,OrmeIM.TheprogressionofchronictuberculosisinthemousedoesnotrequiretheparticipationofBlymphocytesorinterleukin-4.ExpGerontol118.LowrieDB,TasconRE,BonatoVL,etal.TherapyoftuberculosisinmicebyDNAvaccination.119.NuermbergerE,TyagiS,WilliamsKN,RosenthalI,BishaiWR,GrossetJH.RifapentineMoxiŖoxacinorDNAVaccineImprovesTreatmentofLatentTuberculosisinaMouseAmJRespirCritCareMed120.VanEdenW,VanDerZeeR,VanKootenP,etal.Balancingtheimmunesystem:Th1andTh2.AnnRheumDis(Suppl2):ii25121.SilvaCL,BonatoVL,Coelho-CasteloAA,etal.Immu-notherapywithplasmidDNAencodingmycobacterialhsp65inassociationwithchemotherapyisamorerapidandefřcientformoftreatmentfortuberculosisinmice.122.RookGA,DhedaK,ZumlaA.DosuccessfultuberculosisvaccinesneedtobeimmunoregulatoryratherthanmerelyTh1-boosting?Vaccine123.WilkinsonK,KonOM,NewtonS,etal.Tcellmonitoringoftreatmentforlatenttuberculosisinfection.In:internationalconferenceonthepathogenesisofmycobac-terialinfections.Stockholm.Sweden:Saltsjo¨baden;June124.OrmeIM,McMurrayDN,BelisleJT.Tuberculosisvaccinedevelopment:recentprogress.TrendsMicrobiol125.LecoeurHF,Truffot-PernotC,GrossetJH.Experimentalshort-coursepreventivetherapyoftuberculosiswithrifampinandpyrazinamide.AmRevRespirDis126.CardonaPJ,AmatI,GordilloS,etal.EltratamientoępicoconRUTIdelainfeccioęnexperimentalMycobacteriumtuberculosisęneseřcazenARTICLEINPRESS P.-J.Cardona ęspedesresistentesalainfeccioęn.AbstractXI.CongresoSociedadEspanėoladeEnfermedadesInfeccioīsayMicro-īaClīnica.Bilbao,Euzkadi,Spain.June2003.127.GuiradoE,GilO,DűęazJ,etal.ImmunotherapywithRUTIisusefulinthetreatmentoflatentinfectionintheexperi-mentalmodeloftuberculosisinguineapigs.In:internationalconferenceonthepathogenesisofmycobac-terialinfections.Stockholm.Sweden:Saltsjo¨baden;June128.MiddlebrookG.Immunologicalaspectsofairborneinfec-tion:reactionstoinhaledantigens.BacteriolRev129.FerebeeSH.Controlledchemoprophylaxistrialsintuber-culosisAgeneralreview.AdvTubercRes ARTICLEINPRESS RUTI:treatmentoflatenttuberculosisi