RESUMO
Chlamydia muridarum and Chlamydia caviae have equivalent growth rates in mouse epithelial cells but only C. muridarum replicates inside mouse macrophages, while C. caviae does not. Macrophages infected with C. muridarum or C. caviae were used to address the hypothesis that the early signaling pathways initiated during infection depend on the fate of chlamydiae in the host cell. Transmission electron microscopy of C. muridarum-infected macrophages showed intact chlamydial elementary bodies and reticulate bodies 2 h postinfection in compact vacuoles. Conversely, in macrophages infected with C. caviae, chlamydiae were observed in large phagocytic vacuoles. Furthermore, C. caviae infections failed to develop into inclusions or produce viable bacteria. Expression of proinflammatory cytokines TNFα, IL-1ß and MMP13 was similar in C. caviae- or C. muridarum-infected macrophages at 3 h postinfection, indicating that chlamydial survival is not required for initiation of these responses. IL-1ß secretion, dependent on inflammasome activation, occurred in C. caviae-infected macrophages despite no chlamydial growth. Conversely, IFNß mRNA was observed only in C. muridarum- but not in C. caviae-infected macrophages. These data demonstrate that differential signaling events are initiated during a productive versus nonproductive chlamydial infection in a macrophage.
Assuntos
Infecções por Chlamydia/metabolismo , Infecções por Chlamydia/microbiologia , Chlamydia/fisiologia , Espaço Intracelular/microbiologia , Macrófagos/metabolismo , Macrófagos/microbiologia , Transdução de Sinais , Animais , Linhagem Celular , Chlamydia/crescimento & desenvolvimento , Chlamydia/ultraestrutura , Infecções por Chlamydia/genética , Infecções por Chlamydia/patologia , Endossomos/metabolismo , Endossomos/ultraestrutura , Regulação da Expressão Gênica , Inflamação/genética , Interleucina-1beta , Macrófagos/patologia , Macrófagos/ultraestrutura , Camundongos Endogâmicos C57BL , NF-kappa B/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
We have previously shown that MyD88 knockout (KO) mice exhibit delayed clearance of Chlamydia muridarum genital infection compared to wild-type (WT) mice. A blunted Th1 response and ineffective suppression of the Th2 response were also observed in MyD88 KO mice. The goal of the present study was to investigate specific mechanisms whereby absence of MyD88 leads to these effects and address the compensatory mechanisms in the genital tract that ultimately clear infection in the absence of MyD88. It was observed that NK cells recruited to the genital tract in MyD88 KO mice failed to produce gamma interferon (IFN-γ) mRNA and protein. This defect was associated with decreased local production of interleukin-17 (IL-17), IL-18, and tumor necrosis factor alpha (TNF-α) but normal levels of IL-12p70. Additionally, recruitment of CD4 T cells to the genital tract was reduced in MyD88 KO mice compared to that in WT mice. Although chronic infection in MyD88 KO mice resulted in oviduct pathology comparable to that of WT mice, increased histiocytic inflammation was observed in the uterine horns. This was associated with increased CCL2 levels and recruitment of macrophages as a potential compensatory mechanism. Further deletion of TLR4-TRIF signaling in MyD88 KO mice, using TLR4/MyD88 double-KO mice, did not further compromise host defense against chlamydiae, suggesting that compensatory mechanisms are Toll-like receptor (TLR) independent. Despite some polarization toward a Th2 response, a Th1 response remained predominant in the absence of MyD88, and it provided equivalent protection against a secondary infection as observed in WT mice.
Assuntos
Chlamydia muridarum , Interferon gama/metabolismo , Células Matadoras Naturais/metabolismo , Monócitos/fisiologia , Fator 88 de Diferenciação Mieloide/metabolismo , Células Th1/fisiologia , Animais , Infecções por Chlamydia/imunologia , Infecções por Chlamydia/microbiologia , Infecções por Chlamydia/patologia , Tubas Uterinas/patologia , Feminino , Regulação da Expressão Gênica , Inflamação/patologia , Interferon gama/genética , Interleucinas/genética , Interleucinas/metabolismo , Camundongos , Camundongos Knockout , Fator 88 de Diferenciação Mieloide/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Linfócitos T/fisiologia , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/metabolismo , Útero/patologia , Vaginite/microbiologiaRESUMO
The value of routine follow-up in secondary care for cancer patients has been widely questioned. Within our network cancer centre for gynaecological malignancies current follow-up protocols have been associated with delays in diagnosing recurrence. The aim of this study was to ascertain general practitioners' (GPs') attitudes and feasibility of randomisation for a pilot randomised controlled trial to evaluate follow-up of patients treated for gynaecological malignancy. There was a 78% response rate to the postal questionnaire; overall, GP attitudes were positive, with randomisation seeming feasible. We await the results from the pilot trial.