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1、MATERIALSANDMETHODS(RoleforMatrixMetalloproteinase9inGranulomaFormationduringPulmonaryMycobacteriumtuberculosisInfection,fromCell,Vol.148,Issue13,July.2000)Mice.Specific-pathogen-freefemale,6-to8-week-old,CBA/JandC57BL/6micewerepurchasedfromTaconic(Germantown,NY),andwild-typeFVBmiceandMMP-9null(
2、MMP-9KO)breedermicewerepurchasedfromJacksonLaboratory(BarHarbor,ME).TheMMP-9KObreedersweremaintainedasacolonyatColoradoStateUniversity.Allmiceweremaintainedunderbarrierconditionswithsterilemousechowandwateradlibitum.Thespecific-pathogen-freenatureofthemousecolonieswasdemonstratedbytestingsentine
3、lanimals,whichwereshowntobenegativefor12knownmousepathogens.AllexperimentalprocedureswereapprovedbytheColoradoStateUniversityAnimalCareandUseCommittee.小鼠无特异病原体，6-8周大的CBA/J与C57BL/6雌性小鼠购自Taconic(Germantown,NY),野生型FVB小鼠与MMP-9null(MMP-9KO)育种小鼠breedermice购自Jackson实验室(BarHarbor,ME)。MMP-9KO育种动物breeders
4、以群体形式asacolony饲养于美国科罗拉多州大学。所有小鼠都隔栏underbarrierconditions喂养并随意喂无菌鼠食和水sterilemousechowandwateradlibitum。检测岗哨动物以证实小鼠群体的无特异病原体特征，结果表明12种已知的小鼠病原体均呈阴性。所有实验步骤均通过美国科罗拉多州大学动物保护使用委员会ColoradoStateUniversityAnimalCareandUseCommittee同意.。InfectionwithM.tuberculosis.MycobacteriumtuberculosisstrainH37Rv,obtaine
5、dfromtheTrudeauMycobacterialCultureCollection,waspassagedthreetimesthroughpellicleandthenstoredfrozenasaseedstock.WorkingstocksweremadebypassagethroughliquidcultureofProskauer-Beckmediumcontaining0.01%Tween80,andaliquotsoftheorganisminmid-log-phasegrowthwerestoredat80°Cuntilneeded.Micewereinfect
6、edusingproceduresdescribedpreviously(4).Briefly,bacterialstocksweredilutedin5mlofsteriledistilledwaterto2x106CFU/mlandplacedinanebulizerattachedtoanairborneinfectionsystem(Glass-Col,TerreHaute,IN).Micewereexposedto40minofaerosol,duringwhichapproximately50to100bacteriaweredepositedinthelungsofeac
7、hanimal.FortheexperimentsinwhichmiceweretreatedwithBB-94(alsoknownasBatimastat),micewereinfectedwith500CFUintratracheally(17).Bacterialloadwasdeterminedbyplatingwhole-organhomogenatesontonutrient7H11agarsupplementedwitholeic
