Compartmental analysis of proteomic biomarkers during intra-uterine infections

这项研究的目的是描述在生殖器支原体上升感染的特定阶段(从绒毛蜕膜到羊膜内模型)母体和胎儿体内生物标志物表达谱(例如IGFBP-1蛋白水解物片段、钙颗粒蛋白B和膜联蛋白II)之间的机制和时间关系。我们的假设是，宫颈阴道液(CVF)、羊水、母血和胎儿血中特定蛋白质组生物标记物的时空特征将替代宫内感染的进展阶段；类似地，在孕产妇治疗干预(抗生素和抗炎药)过程中生物标记物表达谱的变化将作为预后指标。胎儿的生理适应(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亲抗生素治疗的反应中进行评估。完整的体外研究将评估微小疟原虫通过完整的绒毛羊膜侵入和穿过细胞并进入羊膜腔的能力。 羊水和宫颈阴道液中IL-1B、TNF-α、IL-6和IL-8的生物标志物的产生率和分泌谱将有助于阐明宫内感染过程中羊水和宫颈阴道液中所观察到的特定生物标志物的组织来源。此外，这些研究将提供有关羊膜或绒毛蜕膜在微小葡萄球菌感染后的炎症反应中所起作用的重要信息。一些特定的终点将被确定，这将有助于我们理解绒毛蜕膜定植与解脲支原体之间的因果联系，以及上升子宫感染早期和晚期的生物学和临床表现。子宫收缩能力、宫颈长度的系列评估(通过超声波和触诊)、羊水中PGE2、PGF2a、细胞因子、白细胞和MMPs的水平将与CVF、羊水、母血和胎儿血中的生物标记物的表达谱相关。将对羊水、血液和胎儿组织进行解脲支原体的定量培养和聚合酶链式反应。

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.