RESUMO
Chlamydophila abortus is a Gram-negative obligate intracellular bacterium that causes infectious abortion in sheep (ovine enzootic abortion, OEA) and humans. Infected placentas recovered from sheep that experience OEA have thickened membranes, contain dense inflammatory cellular infiltrates and show evidence of intravascular thrombosis. Despite widespread inflammation, chlamydial multiplication is restricted to the chorionic trophoblast cells. To investigate the potential role of trophoblast in the initiation and propagation of placental inflammation during OEA, the AH-1 ovine trophoblast cell line was experimentally infected with C. abortus and analysed for the release of pro-inflammatory mediators. C. abortus was found to induce the release of both tumour necrosis factor-alpha (TNFalpha) and CXCL8 (interleukin-8) from AH-1 cells in a dose- and time-dependent manner. Ultra-violet (UV)-killed organisms did not elicit this profile, indicating that intracellular multiplication of C. abortus was required for release of these pro-inflammatory mediators. Exposure of AH-1 cells to recombinant ovine TNFalpha alone resulted in the release of CXCL8, suggestive of a self-propagating inflammatory cytokine and chemokine cascade. These data indicate a primary role for trophoblast in the initiation and propagation of placental inflammation during chlamydial abortion.
Assuntos
Aborto Animal/imunologia , Infecções por Chlamydophila/veterinária , Chlamydophila/imunologia , Interleucina-8/metabolismo , Complicações Infecciosas na Gravidez/veterinária , Trofoblastos/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Animais , Linhagem Celular , Proliferação de Células , Chlamydophila/crescimento & desenvolvimento , Infecções por Chlamydophila/imunologia , Infecções por Chlamydophila/patologia , Infecções por Chlamydophila/fisiopatologia , Relação Dose-Resposta Imunológica , Feminino , Homeostase/imunologia , Inflamação/imunologia , Interleucina-8/genética , Interleucina-8/imunologia , Gravidez , Complicações Infecciosas na Gravidez/imunologia , Complicações Infecciosas na Gravidez/patologia , Complicações Infecciosas na Gravidez/fisiopatologia , Ovinos , Trombose/imunologia , Trofoblastos/imunologia , Trofoblastos/microbiologia , Trofoblastos/patologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologiaRESUMO
BACKGROUND: Chlamydiae produce a set of proteins, termed Inc proteins, that are localized to the inclusion membrane and exposed to the host cell cytosol. Little information exists regarding the interaction of Inc proteins with the eukaryotic cell. To examine these interactions, Vaccinia virus vectors and mammalian plasmid-based systems were used to express inc genes in mammalian cells. RESULTS: Cells transfected with plasmids expressing Chlamydophila caviae incA were not productively infected by C. caviae. Expression of C. caviae incA also reduced inclusion formation by Chlamydia trachomatis, but not to the degree seen for C. caviae. Chlamydia trachomatis incA did not block development of either C. trachomatis or C. caviae. Deletion mutagenesis was used to demonstrate that plasmids encoding either the amino or carboxy-terminal regions of the protein, as well as the changing of a single amino acid within IncA (serine 17) could not block C. caviae infection. Immunoblot analysis of truncated IncA in a Vaccinia virus system provided evidence that serine 17 of C. caviae IncA is a target for phosphorylation. CONCLUSIONS: These experiments provide insight into the interaction of Inc proteins with the host cell and introduce a model system where these interactions can be explored further.
Assuntos
Proteínas de Bactérias/genética , Chlamydophila/genética , Células HeLa/microbiologia , Fosfoproteínas/genética , Animais , Proteínas de Bactérias/biossíntese , Células CHO , Linhagem Celular , Linhagem Celular Tumoral , Chlamydia trachomatis/genética , Chlamydia trachomatis/crescimento & desenvolvimento , Chlamydophila/crescimento & desenvolvimento , Cricetinae , Cricetulus , Citoplasma/química , Citoplasma/metabolismo , Citoplasma/microbiologia , Vetores Genéticos/biossíntese , Células HeLa/química , Células HeLa/metabolismo , Células HeLa/virologia , Humanos , Proteínas de Membrana/biossíntese , Proteínas de Membrana/genética , Modelos Genéticos , Fosfoproteínas/biossíntese , Transfecção/métodos , Vaccinia virus/genéticaRESUMO
The BeWo trophoblast cell line does not constitutively express the tryptophan degrading enzyme indolamine 2,3-dioxygenase (IDO), nor can IDO expression be induced by gamma interferon. This correlates with the inability of BeWo cells to control the growth of Chlamydophila abortus, in contrast to effects observed in HeLa cells treated with gamma interferon.