title: Infection of epithelial cells with Chlamydia trachomatis inhibits TNF-induced apoptosis at the level of receptor internalization while leaving non-apoptotic TNF-signalling intact.
abstract: Chlamydia trachomatis is an obligate intracellular bacterial pathogen of medical importance. C. trachomatis develops inside a membranous vacuole in the cytosol of epithelial cells but manipulates the host cell in numerous ways. One prominent effect of chlamydial infection is the inhibition of apoptosis in the host cell, but molecular aspects of this inhibition are unclear. Tumour necrosis factor (TNF) is a cytokine with important roles in immunity, which is produced by immune cells in chlamydial infection and which can have pro-apoptotic and non-apoptotic signalling activity. We here analysed the signalling through TNF in cells infected with C. trachomatis. The pro-apoptotic signal of TNF involves the activation of caspase-8 and is controlled by inhibitor of apoptosis proteins. We found that in C. trachomatis-infected cells, TNF-induced apoptosis was blocked upstream of caspase-8 activation even when inhibitor of apoptosis proteins were inhibited or the inhibitor of caspase-8 activation, cFLIP, was targeted by RNAi. However, when caspase-8 was directly activated by experimental over-expression of its upstream adapter Fas-associated protein with death domain, C. trachomatis was unable to inhibit apoptosis. Non-apoptotic TNF-signalling, particularly the activation of NF-κB, initiates at the plasma membrane, while the activation of caspase-8 and pro-apoptotic signalling occur subsequently to internalization of TNF receptor and the formation of a cytosolic signalling complex. In C. trachomatis-infected cells, NF-κB activation through TNF was unaffected, while the internalization of the TNF-TNF-receptor complex was blocked, explaining the lack of caspase-8 activation. These results identify a dichotomy of TNF signalling in C. trachomatis-infected cells: Apoptosis is blocked at the internalization of the TNF receptor, but non-apoptotic signalling through this receptor remains intact, permitting a response to this cytokine at sites of infection.
authors.affiliation: Institute of Medical Microbiology and Hygiene, University Medical Centre Freiburg, Hermann-Herder-Str. 11, D-79104, Freiburg, Germany.
authors.forename: Collins
authors.initials: C
authors.lastname: Waguia Kontchou
authors.affiliation: Institute of Medical Microbiology and Hygiene, University Medical Centre Freiburg, Hermann-Herder-Str. 11, D-79104, Freiburg, Germany.
authors.forename: Tina
authors.initials: T
authors.lastname: Tzivelekidis
authors.affiliation: Institute of Medical Microbiology and Hygiene, University Medical Centre Freiburg, Hermann-Herder-Str. 11, D-79104, Freiburg, Germany.
authors.forename: Ian E
authors.initials: IE
authors.lastname: Gentle
authors.affiliation: Institute of Medical Microbiology and Hygiene, University Medical Centre Freiburg, Hermann-Herder-Str. 11, D-79104, Freiburg, Germany. georg.haecker@uniklinik-freiburg.de.
authors.forename: Georg
authors.initials: G
authors.lastname: Häcker
mesh_terms: D017209:Apoptosis; D053148:Caspase 3 / Q000378:metabolism; D053181:Caspase 8 / Q000378:metabolism; D002692:Chlamydia trachomatis / Q000502:physiology*; D004847:Epithelial Cells / Q000382:microbiology / Q000502:physiology*; D057809:HEK293 Cells; D006367:HeLa Cells; D054884:Host-Pathogen Interactions; D006801:Humans; D016328:NF-kappa B / Q000378:metabolism; D021381:Protein Transport; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000378:metabolism*; D015398:Signal Transduction; D014409:Tumor Necrosis Factor-alpha / Q000502:physiology*; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: C494826:BAX protein, human; D016328:NF-kappa B; D047888:Receptors, Tumor Necrosis Factor, Type I; C439524:TNF protein, human; D014409:Tumor Necrosis Factor-alpha; D051028:bcl-2-Associated X Protein; C505407:CASP3 protein, human; C505530:CASP8 protein, human; D053148:Caspase 3; D053181:Caspase 8