Uses Host Triacylglycerol to Accumulate Lipid Droplets and Acquires a Dormancy-Like Phenotype in Lipid-Loaded Macrophages
Two billion people are latently infected with Mycobacterium tuberculosis (Mtb). Mtb-infected macrophages are likely to be sequestered inside the hypoxic environments of the granuloma and differentiate into lipid-loaded macrophages that contain triacylglycerol (TAG)-filled lipid droplets which may provide a fatty acid-rich host environment for Mtb. We report here that human peripheral blood monocyte-derived macrophages and THP-1 derived macrophages incubated under hypoxia accumulate Oil Red O-staining lipid droplets containing TAG. Inside such hypoxic, lipid-loaded macrophages, nearly half the Mtb population developed phenotypic tolerance to isoniazid, lost acid-fast staining and accumulated intracellular lipid droplets. Dual-isotope labeling of macrophage TAG revealed that Mtb inside the lipid-loaded macrophages imports fatty acids derived from host TAG and incorporates them intact into Mtb TAG. The fatty acid composition of host and Mtb TAG were nearly identical suggesting that Mtb utilizes host TAG to accumulate intracellular TAG. Utilization of host TAG by Mtb for lipid droplet synthesis was confirmed when fluorescent fatty acid-labeled host TAG was utilized to accumulate fluorescent lipid droplets inside the pathogen. Deletion of the Mtb triacylglycerol synthase 1 (tgs1) gene resulted in a drastic decrease but not a complete loss in both radiolabeled and fluorescent TAG accumulation by Mtb suggesting that the TAG that accumulates within Mtb is generated mainly by the incorporation of fatty acids released from host TAG. We show direct evidence for the utilization of the fatty acids from host TAG for lipid metabolism inside Mtb. Taqman real-time PCR measurements revealed that the mycobacterial genes dosR, hspX, icl1, tgs1 and lipY were up-regulated in Mtb within hypoxic lipid loaded macrophages along with other Mtb genes known to be associated with dormancy and lipid metabolism.
Vyšlo v časopise:
Uses Host Triacylglycerol to Accumulate Lipid Droplets and Acquires a Dormancy-Like Phenotype in Lipid-Loaded Macrophages. PLoS Pathog 7(6): e32767. doi:10.1371/journal.ppat.1002093
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1002093
Souhrn
Two billion people are latently infected with Mycobacterium tuberculosis (Mtb). Mtb-infected macrophages are likely to be sequestered inside the hypoxic environments of the granuloma and differentiate into lipid-loaded macrophages that contain triacylglycerol (TAG)-filled lipid droplets which may provide a fatty acid-rich host environment for Mtb. We report here that human peripheral blood monocyte-derived macrophages and THP-1 derived macrophages incubated under hypoxia accumulate Oil Red O-staining lipid droplets containing TAG. Inside such hypoxic, lipid-loaded macrophages, nearly half the Mtb population developed phenotypic tolerance to isoniazid, lost acid-fast staining and accumulated intracellular lipid droplets. Dual-isotope labeling of macrophage TAG revealed that Mtb inside the lipid-loaded macrophages imports fatty acids derived from host TAG and incorporates them intact into Mtb TAG. The fatty acid composition of host and Mtb TAG were nearly identical suggesting that Mtb utilizes host TAG to accumulate intracellular TAG. Utilization of host TAG by Mtb for lipid droplet synthesis was confirmed when fluorescent fatty acid-labeled host TAG was utilized to accumulate fluorescent lipid droplets inside the pathogen. Deletion of the Mtb triacylglycerol synthase 1 (tgs1) gene resulted in a drastic decrease but not a complete loss in both radiolabeled and fluorescent TAG accumulation by Mtb suggesting that the TAG that accumulates within Mtb is generated mainly by the incorporation of fatty acids released from host TAG. We show direct evidence for the utilization of the fatty acids from host TAG for lipid metabolism inside Mtb. Taqman real-time PCR measurements revealed that the mycobacterial genes dosR, hspX, icl1, tgs1 and lipY were up-regulated in Mtb within hypoxic lipid loaded macrophages along with other Mtb genes known to be associated with dormancy and lipid metabolism.
Zdroje
1. WHO 2009 Global tuberculosis control: a short update to the 2009 report. “WHO /HTM/TB/2009.426”. World Health Organization
2. GomezJEJ.D. McKinney 2004 M. tuberculosis persistence, latency, and drug tolerance. Tuberculosis (Edinb) 84 29 44
3. ZahrtTC 2003 Molecular mechanisms regulating persistent Mycobacterium tuberculosis infection. Microbes Infect 5 159 167
4. ZhangY 2004 Persistent and dormant tubercle bacilli and latent tuberculosis. Front Biosci 9 1136 1156
5. RussellDG 2007 Who puts the tubercle in tuberculosis? Nat Rev Microbiol 5 39 47
6. PeyronPVaubourgeixJPoquetYLevillainFBotanchC 2008 Foamy macrophages from tuberculous patients' granulomas constitute a nutrient-rich reservoir for M. tuberculosis persistence. PLoS Pathog 4: e1000204. doi:1000210.1001371/ journal.ppat.1000204
7. HunterRLJagannathCActorJK 2007 Pathology of postprimary tuberculosis in humans and mice: contradiction of long-held beliefs. Tuberculosis (Edinb) 87 267 278
8. BlochHSegalW 1956 Biochemical differentiation of Mycobacterium tuberculosis grown in vivo and in vitro. J Bacteriol 72 132 141
9. KornbergHLBeeversH 1957 A mechanism of conversion of fat to carbohydrate in castor beans. Nature 180 35 36
10. Munoz-EliasEJMcKinneyJD 2005 Mycobacterium tuberculosis isocitrate lyases 1 and 2 are jointly required for in vivo growth and virulence. Nat Med 11 638 644
11. DanielJDebCDubeyVSD.SirakovaTAbomoelakB 2004 Induction of a novel class of diacylglycerol acyltransferases and triacylglycerol accumulation in Mycobacterium tuberculosis as it goes into a dormancy-like state in culture. J Bacteriol 186 5017 5030
12. BurdonKL 1946 Fatty material in bacteria and fungi revealed by staining dried, fixed slide preparations. J Bateriol 52 665 678
13. GartonNJChristensenHMinnikinDEAdegbolaRABarerMR 2002 Intracellular lipophilic inclusions of mycobacteria in vitro and in sputum. Microbiology 148 2951 2958
14. SirakovaTDDubeyVSDebCDanielJA.KorotkovaT 2006 Identification of a diacylglycerol acyltransferase gene involved in accumulation of triacylglycerol in Mycobacterium tuberculosis under stress. Microbiology 2717 2725
15. DebCLeeCMDubeyVSDanielJAbomoelakB 2009 A novel in vitro multiple-stress dormancy model for Mycobacterium tuberculosis generates a lipid-loaded, drug-tolerant, dormant pathogen. PLoS One 4 e6077 doi:6010.1371/journal.pone.0006077
16. GartonNJWaddellSJSherrattALLeeSMSmithRJ 2008 Cytological and transcript analyses reveal fat and lazy persister-like bacilli in tuberculous sputum. PLoS Med 5: e75 0001-0012
17. ReedMBGagneuxSDeriemerKSmallPMCEB3rd 2007 The W-Beijing lineage of Mycobacterium tuberculosis overproduces triglycerides and has the DosR dormancy regulon constitutively upregulated. J Bacteriol 189 2583 2589
18. BoströmPMagnussonBSvenssonPAWiklundOBorénJ 2006 Hypoxia converts human macrophages into triglyceride-loaded foam cells. Arterioscler Thromb Vasc Biol 26 1871 1876
19. ViaLELinPLRaySMCarrilloJAllenSS 2008 Tuberculous granulomas are hypoxic in guinea pigs, rabbits, and nonhuman primates. Infect Immun 76 2333 2240
20. OttoCMBaumgardnerJE 2001 Effect of culture pO2 on macrophage (RAW 264.7) nitric oxide production. Am J Physiol Cell Physiol 280 C280 287
21. RobinsonMABaumgardnerJEGoodVPOttoCM 2008 Physiological and hypoxic O2 tensions rapidly regulate NO production by stimulated macrophages. Am J Physiol Cell Physiol 294 C1079 1087
22. JamesPEGrinbergOYMichaelsGSwartzHM 1995 Intraphagosomal oxygen in stimulated macrophages. J Cell Physiol 163 241 247
23. NeyrollesOHernández-PandoRPietri-RouxelFFornèsPTailleuxL 2006 Is adipose tissue a place for Mycobacterium tuberculosis persistence? PLoS One 1 e43 doi:10.1371/journal.pone.0000043
24. NykaW 1974 Studies on the effect of starvation on mycobacteria. Infect Immun 9 843 850
25. SeilerPUlrichsTBandermannSPradlLJörgS 2003 Cell-wall alterations as an attribute of Mycobacterium tuberculosis in latent infection. J Infect Dis 188 1326 1331
26. D'AvilaHMeloRCParreiraGGWerneck-BarrosoECastro-Faria-NetoHC 2006 Mycobacterium bovis bacillus Calmette-Guérin induces TLR2-mediated formation of lipid bodies: intracellular domains for eicosanoid synthesis in vivo. J Immunol 176 3087 3097
27. MurphyDJBrownJR 2007 Identification of gene targets against dormant phase Mycobacterium tuberculosis infections. BMC Infect Dis 7 84
28. DebCDanielJSirakovaTDAbomoelakBDubeyVS 2006 A novel lipase belonging to the hormone-sensitive lipase family induced under starvation to utilize stored triacylglycerol in Mycobacterium tuberculosis. J Biol Chem 281 3866 3875
29. CosmaCLShermanDRRamakrishnanL 2003 The secret lives of the pathogenic mycobacteria. Annu Rev Microbiol 57 641 676
30. FriedenTRSterlingTRMunsiffSSWattCJDyeC 2003 Tuberculosis. Lancet 362 887 899
31. BarryCEBoshoffHIDartoisVDickTEhrtS 2009 The spectrum of latent tuberculosis: rethinking the biology and intervention strategies. Nat Rev Microbiol 7 845 855
32. HaapanenJHKassIGensiniGMiddlebrookG 1959 Studies on the gaseous content of tuberculous cavities. Am Rev Respir Dis 80 1 5
33. WayneLGHayesLG 1996 An in vitro model for sequential study of shiftdown of Mycobacterium tuberculosis through two stages of nonreplicating persistence. Infect Immun 64 2062 2069
34. BankaCLBlackASDyerCACurtissLK 1991 THP-1 cells form foam cells in response to coculture with lipoproteins but not platelets. J Lipid Res 32 35 43
35. RiendeauCJKornfeldH 2003 THP-1 cell apoptosis in response to mycobacterial infection. Infect Immun 71 254-259
36. StokesRWDoxseeD 1999 The receptor-mediated uptake, survival, replication, and drug sensitivity of Mycobacterium tuberculosis within the macrophage-like cell line THP-1: a comparison with human monocyte-derived macrophages. Cell Immunol 197 1 9
37. JamesPGrinbergOSwartzH 1998 Superoxide production by phagocytosing macrophages in relation to the intracellular distribution of oxygen. J Leukoc Biol 64 78 84
38. MorNSimonBMezoNHeifetsL 1995 Comparison of activities of rifapentine and rifampin against Mycobacterium tuberculosis residing in human macrophages. Antimicrob Agents Chemother 39 2073 2077
39. RoiniotisJDinhHMasendyczPTurnerAElsegoodCL 2009 Hypoxia prolongs monocyte/macrophage survival and enhanced glycolysis is associated with their maturation under aerobic conditions. J Immunol 182 7974 7981
40. LowKShuiGNatterKYeoWKKohlweinSD 2010 Lipid droplet-associated proteins are involved in the biosynthesis and hydrolysis of triacylglycerol in Mycobacterium bovis bacillus calmette-guerin. J Biol Chem 285 21662 21670
41. KikuchiSRainwaterDLKolattukudyPE 1992 Purification and characterization of an unusually large fatty acid synthase from Mycobacterium tuberculosis var. bovis BCG. Arch Biochem Biophys 295 318 326
42. MattosKADAHRodriguesLSOliveiraVGSarnoEN 2010 Lipid droplet formation in leprosy: Toll-like receptor-regulated organelles involved in eicosanoid formation and Mycobacterium leprae pathogenesis. J Leukoc Biol 87 371 384
43. MishraKCChastellierCdNarayanaYBifaniPBrownAK 2008 Functional role of the PE domain and immunogenicity of the Mycobacterium tuberculosis triacylglycerol hydrolase LipY. Infect Immun 76 127 140
44. CascioferroADeloguGColoneMSaliMStringaroA 2007 PE is a functional domain responsible for protein translocation and localization on mycobacterial cell wall. Mol Microbiol 66 1536 1547
45. FontánPArisVGhannySSoteropoulosPSmithI 2008 Global transcriptional profile of Mycobacterium tuberculosis during THP-1 human macrophage infection. Infect Immun 76 717 725
46. PuissegurMPBotanchCDuteyratJLDelsolGCarateroC 2004 An in vitro dual model of mycobacterial granulomas to investigate the molecular interactions between mycobacteria and human host cells. Cell Microbiol 6 423 433
47. MartinSPartonRG 2006 Lipid droplets: a unified view of a dynamic organelle. Nat Rev Mol Cell Biol 7 373 378
48. YoungDALoweLDClarkSC 1990 Comparison of the effects of IL-3, granulocyte-macrophage colony-stimulating factor, and macrophage colony-stimulating factor in supporting monocyte differentiation in culture. Analysis of macrophage antibody-dependent cellular cytotoxicity. J Immunol 145 607 615
49. AkagawaKS 2002 Functional heterogeneity of colony-stimulating factor-induced human monocyte-derived macrophages. Int J Hematol 76 27 34
50. DolganovGMWoodruffPGNovikovAAZhangYFerrandoRE 2001 A novel method of gene transcript profiling in airway biopsy homogenates reveals increased expression of a Na+-K+-Cl- cotransporter (NKCC1) in asthmatic subjects. Genome Res 11 1473 1483
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Hygiena a epidemiológia Infekčné lekárstvo LaboratóriumČlánok vyšiel v časopise
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