Immune Control of Group B Streptococcal Placental

项目摘要/摘要 羊膜内感染和炎症仍然是早产、死产和新生儿的重要原因。 发病率和死亡率。这项建议的目标是定义早期母体和胎盘免疫 对解决母胎界面细菌感染至关重要的反应。澄清： 发生在母胎交界处的免疫事件在人类妊娠中是复杂的 母体和胎儿隔间无法进入，这也限制了我们对 入侵生物体的性质和指导细菌清除的细胞类型。我们已经克服了这些困难 使用独特的长期插管怀孕的非人类灵长类动物(NHP)模型进行挑战 模仿人类怀孕。在这项建议中，我们将阐明早期免疫机制，导致 细菌清除和确定保护胎儿免受感染的关键免疫细胞类型和宿主防御网络 侵袭性细菌感染。我们将使用已建立的GBS感染的NHP模型来研究细菌 使用创新方法清除母胎界面，包括：1)多维流动 细胞术定量母胎交界处的免疫细胞群，2)单细胞RNA-Seq 生成细胞类型和调控基因网络的转录图谱，3)反相蛋白质阵列 分析信令级联和主机转换网络以及4)复杂的计算建模 将临床元数据(例如细菌负荷、子宫活动峰值)与单细胞RNA-Seq和蛋白质阵列联系起来 数据。因此，拟议的目标将确定免疫反应和宿主的时间和空间性质 转录和翻译网络对母胎界面细菌的清除至关重要 怀孕了。

PROJECT SUMMARY/ABSTRACT Intra-amniotic infection and inflammation remain a significant cause of preterm birth, stillbirth and neonatal morbidity and mortality. The objective of this proposal is to define early maternal and placental immune responses that are critical for resolution of bacterial infections at the maternal-fetal interface. Elucidation of immune events occurring at the maternal-fetal interface in human pregnancy is complicated by the inaccessibility of maternal and fetal compartments, which also imposes limitations on our understanding of the nature of the invading organism and cell types directing bacterial clearance. We have overcome these challenges by using a unique chronically catheterized pregnant nonhuman primate (NHP) model that closely emulates human pregnancy. In this proposal, we will elucidate early immune mechanisms that result in bacterial clearance and define the key immune cell-types and host defense networks that protect the fetus from invasive bacterial infections. We will use the established NHP model of GBS infection to study bacterial clearance at the maternal-fetal interface using innovative methods including: 1) multidimensional flow cytometry to quantitate immune cell populations at the maternal-fetal interface, 2) single cell RNA-Seq to generate a transcriptional map of cell types and regulatory gene networks, 3) reverse phase protein array to analyze signalling cascades and host translational networks and 4) sophisticated computational modelling to link clinical metadata (e.g. bacterial burden, peak uterine activity) with single cell RNA-Seq and protein array data. The proposed aims will thus establish the temporal and spatial nature of immune responses and host transcriptional and translational networks essential for bacterial clearance at the maternal-fetal interface during pregnancy.