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TUBERCULOSIS
Puedeafectarcualquierrganoysistemaenformaprogresivay crnicaporloqueelcuadroclnicoseencuentrarelacionado conelsitioafectado. Sepresentaprincipalmenteanivelpulmonarypuede diseminarseaotrossitios. LaaparicindelVIHhacausadoaumentoenlasformas extrapulmonares quesondifcilesdeDx.
COMOSECONTAGIALATUBERCULOSIS?
Diseminacinaotrosrganos
FORMASDELAENFERMEDAD
PorquestanexitosoMycobacterium?
Parsitoobligado,intracelular
Some mycobacteria are degraded within macrophages. Infected macrophages were processed for transmission electron microscopy at 24 (A), 48 (B), 72 (C), 96 (D), and 120 (E) hours postinfection. Degraded, unrecognizable mycobacteria were labeled with immunogold particles (AE). Healthy Mycobacteria at 72 hours postinfection were seen in (F). Magnification: 104,000 (AC), 104,000 (D), 165,000 (E), 88,400 (F).
Mycobacterium tuberculosis
BacilodescritoporprimeravezporRobertoKock en1882 Bacilodelgado,cidoalcoholresistente Crecimientolento(duplicacin520hrs) Altocontenidodelpidos (paredgruesa)
Aislamiento de M. tuberculosis
Muestra +(NAOH,Citrato deNa,NAcetil cisteina ) 15min Centrifugar Decantar ylavar conPBS AjustarelpH=7.0
Aparicindecolonias(48semanas)
Mycobacterium tuberculosispresentacrecimientoexcelente.Lasclulastiendenaformar agregadoscompactosporlanaturalezahidrofbica delapared Lacoloniassongrandes,secas,amarillentasygranulares
MORFOLOGIACOLONIAL
IdentificacindeMycobacterium tuberculosis
Loscidosmiclicos lesconfieren lapropiedadderesistirla decoloracinconalcoholcido, despusdelatincincon colorantesbsicos(fucsina fenicada). Serequierecalentamientopara queelcoloranteatraviesela paredbacteriana. Posteriormenteselavancon alcoholcidoyseadicionaun colorantedecontraste(azulde metileno).
Tincin ZhielNeelsen
IdentificacindeMycobacterium tuberculosis
Loscidosmiclicos tienen afinidadporlosfluorocromos auramina yrodamina.Estos colorantessefijanalas bacterias,queaparecende coloramarilloonaranja. Elpermanganatodepotasio, empleadocomocontraste, evitalafluorescencia inespecfica. Todoslosmicroorganismos cidoalcoholresistentesson positivos.
Tincin AuraminaRodamina
QUETIENEDEESPECIAL?
Bacterias Grampositivas dealtocontenido deGC,aerobia ymesofila
Reino:Bacteria Phylum:Actinobacteria Clase:Actinobacteridae Orden:Actinomycetales Corynebacterineae Familia:Mycobacteriaceae Gnero:Mycobacterium Especie:Mycobacterium tuberculosis
Elgnero Mycobacterium puede infectar adiversas especies deanimales como:aves, peces ,roedores yhumanos.
MORFOLOGIACELULAR
H37Ra
CDC1551
(Matrixextracelular)
Fig. 3. MTP binds to laminin. (A) ELISAbased assay to measure the binding of increasing concentrations of fibronectin (), laminin (), and collagen type IV ( ) to MTPcoated plates. Binding was detected by using either rabbit antifibronectin, antilaminin, or mouse anticollagen type IV.
Fig. 3. MTP binds to laminin. (B) Flow cytometry demonstration of binding of laminin to MTPproducing M. tuberculosis but not to mtp mutants. Binding was detected by using either rabbit antifibronectin, antilaminin, or mouse anticollagen type IV. Data are presented from triplicate experiments
Fig. 4. MTP are produced during human TB infection. (A) IF showing that MTP incubated with TB patient sera produced fluorescent MTP fibers. (B) No reaction was observed when MTP was incubated with sera from healthy donors. (Original magnification: 1,000.) (C) Sera from TB patients (n=36) and from healthy donors (n=5) were tested for antiMTP IgG antibodies by ELISA. Most of the patients sera (60%) showed a significant IgG titer against immobilized MTP fibers. Results obtained at sera diluted 1:3,200 are shown. The horizontal line represents the cutoff value of two times the average ELISA reading of healthy control sera. All of the sera positive by ELISA produced a positive IF reaction as shown in A.
CAPSULA
Capsulaestacompuestadecarbohidratos (glicanos yarabinomananos) Glucosa Manosa Arabinosa Lpidos Protenas(19kDa,complejo85) Sehademostradoquelacapsulalimitala interaccinconelmacrfagodurantelas etapastempranasdelainfeccin. Sinembargosehareportadoquealgunos polisacridosdelacapsulaseunenal macrfagovareceptor3decomplemento, evitandolaactivacindelmacrfago
QUELAHACEEXITOSA? COMPARACINCONOTRASBACTERIAS
SchematicRepresentationofBacterialCellWalls
Grampositive bacteria have a thick layer of PG. Lipoteichoic acids and lipoproteins are embedded in this cell wall. The cell wall of Gramnegative bacteria is characterized by the presence of LPS. Mycobacteria have a thick hydrophobic layer containing mycolyl arabinogalactan and dimycolate, in addition to a lipid bilayer and a PG layer. Lipoarabinomannan (LAM) is a major cellwall associated glycolipid. Lipoproteins are common structures for various types of bacteria.
PEPTIDOGLICANOS
Estructuradelaunidadbsicadelospepidoglicanos
DAP
LISINA
COMPOSICINDELAPAREDGRAM
UNIONNOCOVALENTEDELACAPADEPEPTIDOGLICANOSPORLIPOPROTEINAS
ESTRUCTURADELLIPOPOLISACARIDO
Lasecuenciadifiereentreespecies,perolasecuencia deloselementosprincipalesesuniforme.
COMPOSICINDELAPAREDGRAM+
ESTRUCTURABASICA DELAC.TEICOICO
Propiedadesdelaparedcelular
Laparedposeeunelevado contenidoenlpidos(5060%)que leconfierenuncarcter hidrofbico Estconstituidaporelcomplejo macromolecularformadopor cidosmiclicoarabinogalactano peptidoglucano (mAGP) Loscidosmiclicos tienende70 80carbonosyselesatribuyeel carcterhidrofbico dela envoltura.
PAREDCELULARCOMPLEJA
cord factor dimycolyltrehalose, sulpholipids and phosphatidylinositol Mannosides andLAM
Schematic representation of the cell envelope of Mycobacterium tuberculosis. The cell wall contains three different covalently linked structures (peptidoglycan ,grey; arabinogalactan, blue and mycolic acids, green).
Acido miclico: un trmino genrico para unos cidos grasos de elevado peso molecular a-ramificados b-hidroxilados que forman la pared de las micobacterias.
Donde R1 es una cadena hidrocarbonada de 20 a 24 C y R2 una estructura compleja de 30 a 60 tomos de carbono que puede contener dobles enlaces, anillos cicloproplicos y funciones oxigenadas.
MOLCULASDELAPAREDCELULAR
LAM(lipoarabinomananos)son secretadosdemacrfagosinfectados conmicobacterias. Afectanlafuncindeclulasvecinasno infectadasylosactivan LAMyLMtienencaractersticasde PAMP(pathogenassociated molecular patterns),molculasexpresadas invariablementeporpatgenos relacionados. PAMPsonligandos dereceptoresdela respuestainmuneinnataexpresadasen losfagocitos.
Lipomannans
Receptoresdelsistemainmuneinvolucradosenelreconocimientode elementosdeparedcelulardeM.tuberculosis
IMPORTANCIADELOSCOMPONENTESDELAPAREDCELULARENLA RESPUESTAINMUNE
IMPORTANCIADELOSCOMPONENTESDELAPAREDCELULARENLA RESPUESTAINMUNE
Abreviaturas:(GPL);Fenolglicolpidos (PGL);Fosfatidilinositil (PI);Fosfatidilinositolmansidos (PIM);Lipoarabinomanana (LAM);Lipoarabinomanana conresiduosterminalesenmanosa (ManLAM);Lipoarabinomanana sinresiduosterminalesde manosa (AraLAM).
ELEMENTOSEXTRACELULARESDEVIRULENCIA
M.tuberculosissintetizapequeasmolculas extracelularesdeorigenproteicoqueestnrelacionadas conlavirulencia. Estosfactoresdevirulenciaestnausentesenlascepas deBCG(Bovis Calmette Guerin). ESAT6yCFP10 Sonsecretadasconstitutivamente,capacesdeatenuarla respuestainmuneinnatavaTLRs
SISTEMADESECRECINPARAESAT6
Modelo delsistema ESX1
SISTEMASDEEXPORTACINDELOSFACTORES DEVIRULENCIA
QUECOMEM.tuberculosis?
Nutrientesquepuedenestardisponiblespara M.tuberculosisenelhospedero
MetabolismogeneraldeM.tuberculosis
LOSLIPIDOSSONLAFUENTEPREFERENCIALDECARBONO
ESTUDIOSINSILICOPUBLICADOSENNature 393,537544(1998)
UTILIZACINDECARBONOENPARIENTESCERCANOS
Crecimiento deMycobacterium envarios carbohidratos Comounica fuente decarbono
M.tuberculosisescapazdeutilizar diversasfuentesdecarbono
4 5 7 8 3 6 1 2 Tiempo de generacin g = t/n . Es el tiempo requerido para que a partir de una clula se formen dos.
Growth of Mtb in minimal medium with different carbon sources. The growth of Mtb in 37C in HB medium containing 1% glucose (circles), 1% trehalose (diamonds), 1% maltose (squares), and 1% GlcNAc (triangles) was measured by determining the OD600 of the cultures. The values are the means of three independent experiments.
INCORPORACINDECARBOHIDRATOSDEPARIENTESCERCANOS
Escapazdeutilizardiversasfuentesdecarbono
M.smegmatis
FIG. 5. Kinetics and inducibility of fructose, glucose, and glycerol uptake. Mycobacterium was grown in Middlebrook 7H9 medium in the presence of 2% glycerol (open circles) or 2% fructose (closed circles). (B) Mtb was grown in the presence of 2% glycerol (open circles), 2% glucose (closed circles), and 2% glycerol plus 2% glucose (closed squares). (C) Mtb was grown 0.4% glucose (open circles) and 0.4% glycerol (closed circles)
GENESINVOLUCRADOSENLAUTILIZACINDECARBOHIDRATOS Mycobacterium
CLASIFICACINDELOSSISTEMASDETRANSPORTE