Immune Control of Group B Streptococcal Placental

项目总结/摘要 羊膜内感染和炎症仍然是早产、死产和新生儿窒息的重要原因。 发病率和死亡率。这项建议的目的是确定早期母体和胎盘免疫 这些反应对于解决母胎界面处的细菌感染至关重要。阐明 在人类妊娠中发生在母胎界面的免疫事件是复杂的 母亲和胎儿室的不可接近性，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 通过使用独特的长期插管妊娠非人灵长类动物（NHP）模型， 模仿人类怀孕在这个提议中，我们将阐明导致 细菌清除和定义关键的免疫细胞类型和宿主防御网络，保护胎儿免受 侵入性细菌感染。我们将利用已建立的GBS感染NHP模型， 使用创新方法清除母胎界面，包括：1）多维流动 流式细胞术以定量母胎界面处的免疫细胞群体，2）单细胞RNA-Seq以 生成细胞类型和调控基因网络的转录图谱，3）反相蛋白阵列， 分析信号级联和宿主翻译网络，以及4）复杂的计算建模， 将临床元数据（例如细菌负荷、峰值子宫活动）与单细胞RNA-Seq和蛋白质阵列相关联 数据因此，所提出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体-胎儿界面的细菌清除至关重要， 怀孕

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.