Compartmental analysis of proteomic biomarkers during intra-uterine infections

本研究提案的目的是表征生物标志物表达谱之间的机制和时间关系（例如，IGFBP-1蛋白水解片段、钙颗粒蛋白B和膜联蛋白II）在生殖器支原体上行感染的确定阶段（从绒毛膜蜕膜到羊膜内模型）的母体和胎儿隔室中。我们假设宫颈阴道液（CVF）、羊水、母体和胎儿血液中特定蛋白质组生物标志物的空间和时间特征将作为子宫内感染进展阶段的替代物;同样，母体治疗干预（抗生素和抗炎剂）期间生物标志物表达谱的变化将作为预后指标。胎儿生理适应（即，心血管血流动力学、呼吸参数、内分泌状态）将在感染的早期和晚期以及对母体抗生素治疗的反应中进行评估。补充的体外研究将评估U.小虫通过完整的绒毛膜羊膜侵入和转胞吞并进入羊膜腔。 特异性生物标志物的产生速率和组成组织层的IL-1B、TNF-α、IL-6和IL-8的分泌谱将阐明在子宫内感染期间在羊水和宫颈阴道液（CVF）中观察到的特异性生物标志物谱的组织来源。此外，这些研究将提供重要的信息，羊膜或绒毛膜蜕膜的贡献，炎症反应后，U。小暴露。一些特定的终点将被确定，这将有助于我们理解绒毛膜蜕膜定植与U。以及上行性子宫感染早期和晚期的生物学和临床表现。子宫收缩力、宫颈长度的系列评估（通过超声和触诊）、PGE 2、PGF 2a、细胞因子、白细胞和MMP的羊水水平将与CVF、羊水和母体和胎儿血液中的生物标志物表达谱相关。将对羊水、血液和胎儿组织进行支原体定量培养和PCR。

The objectives of this research proposal are to characterize the mechanistic and temporal relationships among biomarker expression profiles (e.g., IGFBP-1 proteolytic fragments, calgranulin B and annexin II) in maternal and fetal compartments during defined stages of ascending infection with genital mycoplasmas (from choriodecidual to intra-amniotic models). It is our hypothesis that spatial and temporal characteristics of specific proteomic biomarkers in cervical vaginal fluid (CVF), amniotic fluid, maternal and fetal blood, will act as surrogates for the stage of progression of intra-uterine infection; similarly, changes in biomarker expression profiles during maternal therapeutic interventions (antibiotic and anti-inflammatory agents), will serve as prognostic indicators. Fetal physiological adaptations (i.e., cardiovascular hemodynamics, respiratory parameters,endocrine status) will be assessed in early and advanced stages of infection, and in response to maternal antibiotic therapy. Complemetary in vitro studies will assess the ability of U. parvum to invade and transcytose through intact chorioamniotic membranes and gain entry into the amniotic cavity. The production rates of specific biomarkers and the secretory profiles of IL-1B, TNF-a, IL-6 and IL-8 by component tissue layers will illuminate the tissue source of specific biomarker profiles observed in the amniotic fluid and cervical vaginal fluid (CVF) during intra-uterine infection. Moreover, these studies will provide important information on the contribution of the amnion or choriodecidua to the inflammatory response following U. parvum exposure. A number of specific endpoints will be ascertained that will aid in our understanding of causal links among choriodecidual colonization with U. parvum and the biologic and clinical manifestations of early and late stages of ascending uterine infection. Uterine contractility, serial assesment of cervical length (by ultrasound and palpation), amniotic fluid levels of PGE2, PGF2a, cytokines, leukocytes and MMPs will be correlated with biomarker expression profiles in the CVF, amniotic fluid and maternal and fetal blood. Quantitative cultures and PCR for ureaplasmas will be performed on amniotic fluid, blood and fetal tissues.