RESUMEN
BACKGROUND: Ascending infections of the female genital tract with bacteria causes pelvic inflammatory disease (PID), preterm labour and infertility. Lipopolysaccharide (LPS) is the main component of the cell wall of Gram-negative bacteria. Innate immunity relies on the detection of LPS by Toll-like receptor 4 (TLR4) on host cells. Binding of LPS to TLR4 on immune cells stimulates secretion of pro-inflammatory cytokines such as IL-6, chemokines such as CXCL1 and CCL20, and prostaglandin E(2). The present study tested the hypothesis that TLR4 on endometrial epithelial and stromal cells is essential for the innate immune response to LPS in the female genital tract. METHODOLOGY/PRINCIPAL FINDINGS: Wild type (WT) mice expressed TLR4 in the endometrium. Intrauterine infusion of purified LPS caused pelvic inflammatory disease, with accumulation of granulocytes throughout the endometrium of WT but not Tlr4(-/-) mice. Intra-peritoneal infusion of LPS did not cause PID in WT or Tlr4(-/-) mice, indicating the importance of TLR4 in the endometrium for the detection of LPS in the female genital tract. Stromal and epithelial cells isolated from the endometrium of WT but not Tlr4(-/-) mice, secreted IL-6, CXCL1, CCL20 and prostaglandin E(2) in response to LPS, in a concentration and time dependent manner. Co-culture of combinations of stromal and epithelial cells from WT and Tlr4(-/-) mice provided little evidence of stromal-epithelial interactions in the response to LPS. CONCLUSIONS/SIGNIFICANCE: The innate immune response to LPS in the female genital tract is dependent on TLR4 on the epithelial and stromal cells of the endometrium.
Asunto(s)
Endometrio/inmunología , Células Epiteliales/inmunología , Lipopolisacáridos/inmunología , Células del Estroma/inmunología , Receptor Toll-Like 4/inmunología , Animales , Citocinas/inmunología , Femenino , Mediadores de Inflamación/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Receptor Toll-Like 4/deficiencia , Receptor Toll-Like 4/genéticaRESUMEN
BACKGROUND: Escherichia coli are widespread in the environment and pathogenic strains cause diseases of mucosal surfaces including the female genital tract. Pelvic inflammatory disease (PID; metritis) or endometritis affects approximately 40% of cattle after parturition. We tested the expectation that multiple genetically diverse E. coli from the environment opportunistically contaminate the uterine lumen after parturition to establish PID. METHODOLOGY/PRINCIPAL FINDINGS: Distinct clonal groups of E. coli were identified by Random Amplification of Polymorphic DNA (RAPD) and Multilocus sequence typing (MLST) from animals with uterine disease and these differed from known diarrhoeic or extra-intestinal pathogenic E. coli. The endometrial pathogenic E. coli (EnPEC) were more adherent and invasive for endometrial epithelial and stromal cells, compared with E. coli isolated from the uterus of clinically unaffected animals. The endometrial epithelial and stromal cells produced more prostaglandin E(2) and interleukin-8 in response to lipopolysaccharide (LPS) purified from EnPEC compared with non-pathogenic E. coli. The EnPEC or their LPS also caused PID when infused into the uterus of mice with accumulation of neutrophils and macrophages in the endometrium. Infusion of EnPEC was only associated with bacterial invasion of the endometrium and myometrium. Despite their ability to invade cultured cells, elicit host cell responses and establish PID, EnPEC lacked sixteen genes commonly associated with adhesion and invasion by enteric or extraintestinal pathogenic E. coli, though the ferric yersiniabactin uptake gene (fyuA) was present in PID-associated EnPEC. Endometrial epithelial or stromal cells from wild type but not Toll-like receptor 4 (TLR4) null mice secreted prostaglandin E(2) and chemokine (C-X-C motif) ligand 1 (CXCL1) in response to LPS from EnPEC, highlighting the key role of LPS in PID. CONCLUSIONS/SIGNIFICANCE: The implication arising from the discovery of EnPEC is that development of treatments or vaccines for PID should focus specifically on EnPEC and not other strains of E. coli.
Asunto(s)
Endometrio/microbiología , Infecciones por Escherichia coli/microbiología , Escherichia coli/fisiología , Enfermedad Inflamatoria Pélvica/microbiología , Animales , Bovinos , Enfermedades de los Bovinos/metabolismo , Enfermedades de los Bovinos/microbiología , Células Cultivadas , ADN Bacteriano/análisis , ADN Bacteriano/genética , Endometritis/inducido químicamente , Endometritis/metabolismo , Endometritis/microbiología , Endometrio/citología , Escherichia coli/clasificación , Escherichia coli/genética , Infecciones por Escherichia coli/metabolismo , Infecciones por Escherichia coli/veterinaria , Femenino , Genotipo , Interacciones Huésped-Patógeno , Interleucina-8/metabolismo , Lipopolisacáridos/metabolismo , Lipopolisacáridos/toxicidad , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Enfermedad Inflamatoria Pélvica/metabolismo , Filogenia , Técnica del ADN Polimorfo Amplificado Aleatorio , Especificidad de la Especie , Receptor Toll-Like 4/genética , Receptor Toll-Like 4/metabolismo , Útero/microbiologíaRESUMEN
Thromboxane A2 (TXA2) is an arachidonate metabolite which is considered to relate to chronic inflammation in atopic diseases characterized by elevated immunoglobulin E productivity. The elevation of immunoglobulin E levels involves many molecules including interleukin-4 (IL-4) and interleukin-4 receptor alpha chain (IL-4R alpha). To assess whether genetic variants of TXA2 receptor, IL-4 and IL-4R alpha genes relate to the elevation of serum immunoglobulin E levels in patients with atopic dermatitis (AD), we conducted an association study of genetic polymorphisms of TXA2 receptor (795C/T), IL-4 (-589C/T), and IL-4R alpha (Ile50Val) in a Japanese population (n = 789). The TXA2 receptor 795TT genotype strongly related to AD with high serum immunoglobulin E concentrations. AD patients with both TXA2 receptor 795TT genotype and the IL-4R alpha Ile50/Ile50 genotype showed the greatest immunoglobulin E concentrations. These results suggest TXA2 receptor polymorphism strongly interacts with IL-4R alpha polymorphism as a major determinant of high serum immunoglobulin E levels in AD.