1. Christiane Brohm26.01.2009 (1) M. tuberculosis profile (2) Infection route (3) Survival strategies of M. tuberculosis

3. Mycobacterium Tuberculosis Profile Family: Mycobacteriaceae Genus: Mycobacterium Feature:waxy coating gram-positive obligate aerobe Disease: Tuberculosis Prevalence: 2 billion worldwide 10% casuality Therapy:antimycobacterial therapy (e.g. isoniazid, rifampicin)

4. Infection Route of M. tuberculosis (1) Inhalation of aerosols containing bacilli (2) Phagocytosis in lung by resident macrophages (3) Lysosomal degradation or survival within macrophage

5. Infection Route of M. tuberculosis - Phagosomal Way - Nature Reviews Immunology 1, 20-30 (October 2001) | doi:10.1038/35095558 (1) Internalization into phagosome → delivery to antigen processing/presentation pathway (2) Activation of T-cells by peptide-loaded MHC class II molecules; mycobacterial lipids presented to γδ T cells → adaptive immune response But: Escape of immune defense mechanisms

6. Survival Strategies of M. tuberculosis (1) Phagocytosis into macrophage → receptor dependent macrophage activation (6) Subversion of macrophage activation → reaction on oxygen/nitrogen (5) Hijacking cellular calcineurin pathway → preventing fusion/degradation (4) Mimicking of host signaling molecules → preventing fusion/degradation (3) Prevention of phagosome-lysosome fusion → no degradation but persistence (2) Establishment of a balance: the granuloma → persistence for long time

7. Bacteria – Host Balance : The Granuloma - T cell activation following presentation of mycobacterial antigens - actively dividing bacilli or „dormant“ state - attenuated immune system can lead to development of tuberculosis

8. Mycobacterial Prevention of Phagosome-Lysosome-Fusion Non-infected cells: Generation of PI3P regulates delivery of phagocytosed cargo to lysosomes PI3P: phosphatidylinositol 3-phosphate LAM: mycobacterial cell-wall component lipoarabinomannan SapM: eukaryotic-like acid phosphatase secreted by M.t. Infected cells: Interference with phagosome-lysosome fusion M. tuberculosis (1) prevents PI3P generation by mycobacterial LAM (2) hydrolizes PI3P on phagosomal membranes by SapM

9. Production of Host-like Signaling Molecules Nature Medicine 13, 282 - 284 (2007) doi:10.1038/nm0307-282 Mycobacterial production of eukaryotic-like serine/threonine kinases - two kinases are soluble and thereby released into the cytoplasm PknG Prevention of phagosome-lysosome fusion Drug target Specific kinase inhibitor interacting with the unique domain of PknG Inhibitors do not need to aquire access to impermeable mycobacterial cell wall

10. Interfering with Host Cell Signaling Hijacking the Calcineurin Pathway TACO, P57, Coronin 1 Host factor that specifically prevents lysosomal delivery. Exclusively presence on phagosomes harboring mycobacteria. Regulation of calcium-dependent signaling processes: Infected macrophages respond with sustained calcium flux dependent on Coronin 1. Activation of calcineurin blocks fusion. Calcineurin blockers (cyclosporin A, FK506) fully block mycobacterial proliferation.

11. Macrophage Activation and Its Subversion by M. tuberculosis Block of fusion only in non-activated macrophages. What happens in activated macrophages? (1) Mycobacterial production of KatG: Inactivation of reactive oxygen (2) Mycobacterial proteasome copes with nitric-oxid stress KatG: mycobacterial catalase-peroxidase

12. Macrophage Activation and Its Subversion by M. tuberculosis Cathelicidin TLR-ligands can cause macrophage activation Phagosome maturation through TLR adaptor MyD88 and p38MAPK Upregulation of vitamin D receptor Induction of cathelicidin (antimicrobial peptide) MyD88: myeloid differentiation factor 88 p38MAPK: mitogen-associated protein kinase p38 protein

13. Macrophage Activation and Its Subversion by M. tuberculosis LAM interferes with macrophage by modulating signaling pathways IFN-γ-mediated gene expression TLR activation MAPK activation Phagosome-lysosome fusion Modulation of bacterial metabolic pathways fatty acids as carbon source within macrophages Sec-dependent secretion pathway (signal-sequence dependent transport) Tat-system (transport of folded molecules) ESX-1 secretion system (transport of virulence factors, escape of bacilli into cytoplasm) LAM: cell-wall component lipoarabinomannan Tat-system: twin-arginine transporter system ESX-1: early secretory antigenic target of 6kD (ESAT-6) system 1

14. Conclusions M. tuberculosis plays hide-and-seek in phagosomes within macrophages and granulomas within the lung. prevents phagosome-lysosome fusion by mimicking and hijacking host signaling pathways. actively reacts and persists in activated macrophages. Prolonged coevolution of M. tuberculosis with its human host resulted in a number of survival strategies.

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