Single-Cell Analysis To Define Protective and Tolerizing Immune Cell Populations in the Human Placenta

单细胞分析确定人胎盘中的保护性和耐受性免疫细胞群

基本信息

批准号：
9978484
负责人：
Martin Prlic
金额：
$ 28.33万
依托单位：
FRED HUTCHINSON CANCER RESEARCH CENTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-07 至 2022-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9978484
关键词：
Activities of Daily Living Agonist Allogenic Antigen Presentation Antigen-Presenting Cells Antigens Autoantigens Autoimmunity Biological Assay Biological Markers Blood Cells Coculture Techniques Color Cytometry Data Decidua Dendritic Cells Disease Event Exposure to Fetus Flow Cytometry Fluorescence Future Gene Expression Profiling Goals Health Heterogeneity Human Immune Immune response Immune system Immunocompetence Immunologics Infection Intervention Link Maintenance Maternal Exposure Measures Mediating Mothers Nature Outcome Pathologic Pathology Phenotype Placenta Play Population Population Heterogeneity Pre-Eclampsia Pregnancy Professional Role Property Proteins Reproduction Role System T-Cell Activation T-Lymphocyte T-Lymphocyte Subsets Testing Time Tissues Transplantation Umbilical Cord Blood Uterus adaptive immune response adverse pregnancy outcome base chemokine cytokine design early pregnancy embryo/fetus antigen exhaustion experimental study fetal fighting high dimensionality improved insight inter-individual variation mouse model offspring pregnancy disorder prevent receptor reproductive response single cell analysis therapeutic target transcriptome

项目摘要

Maternal immune adaptation to pregnancy requires maintenance of immune competence with concurrent specific tolerance of the semi-allogeneic fetus. Immune maladaptation has been associated with several adverse pregnancy outcomes, with both short- and long-term impact on maternal and offspring health. Maternal exposure to fetal-placental antigens during pregnancy is well-established; however, many questions remain about the nature of both antigen presentation and the resultant adaptive immune response. Eliciting a de novo adaptive immune response requires the activation of professional antigen presenting cells (APCs), especially dendritic cells (DCs). Importantly, DCs can also induce T cell tolerance and thus play a critical role in avoiding unwanted immune responses. This has been typically studied in the context of preventing responses against self-antigens and is also relevant for protection of the fetus from maternal rejection. It remains unknown how APCs in the human placenta and decidua provide protection against infection and at the same time prevent immune responses against the fetus. A major roadblock to understanding protection and tolerance in human reproduction is the limited information about the functional profile of these APCs, and their inter-individual variation. We propose to define human APC subsets in maternal blood, placenta, decidua, and umbilical cord blood to test the hypothesis that APCs involved in human reproduction consist of several subsets that are functionally specialized to contribute to either protection or tolerance. To accomplish this, we will take advantage of recent technological advances in the field of single-cell analysis, namely high- dimensional cytometry and single-cell gene expression analysis to dissect the functional heterogeneity within this immune compartment. Our lab has developed and validated multiple 28-color fluorescence cytometry assays, which interrogate cell phenotype, activation, and functional capacity (i.e. expression of exhaustion markers, inhibitory receptors, co-stimulatory molecules). We will employ these assays to determine the composition and capabilities of cells directly ex vivo. These experiments will provide critical insight to define functional specialization of DCs in healthy human reproductive tissues during pregnancy and also provide the basis to study pathologies such as preeclampsia in the future.

母体对妊娠的免疫适应需要维持免疫能力，同时半同种异体胎儿的特异性耐受性。免疫适应不良与多种因素有关不良妊娠结局，对孕产妇和后代健康产生短期和长期影响。母亲在怀孕期间接触胎儿胎盘抗原已是公认的；然而，许多问题仍然是关于抗原呈递和由此产生的适应性免疫反应的本质。引出一个从头适应性免疫反应需要激活专业抗原呈递细胞（APC），特别是树突状细胞（DC）。重要的是，DC 还可以诱导 T 细胞耐受，从而在避免不需要的免疫反应方面发挥关键作用。这通常是在预防针对自身抗原的反应的背景下进行研究的，并且也与保护胎儿免受母体排斥有关。目前尚不清楚人类胎盘和蜕膜中的 APC 如何提供针对感染的保护，同时防止针对胎儿的免疫反应。了解人类生殖中的保护和耐受的一个主要障碍是关于这些 APC 的功能概况及其个体间变异的信息有限。我们建议定义母血、胎盘、蜕膜和脐带血中的人类 APC 亚群，以检验以下假设：参与人类生殖的 APC 由几个功能专门化的亚群组成，有助于保护或耐受。为了实现这一目标，我们将利用单细胞分析领域的最新技术进步，即高三维细胞计数和单细胞基因表达分析来剖析内部的功能异质性这个免疫室。我们实验室开发并验证了多种28色荧光细胞仪检测，询问细胞表型、激活和功能能力（即耗尽的表达）标记物、抑制性受体、共刺激分子）。我们将利用这些测定来确定直接离体细胞的组成和功能。这些实验将为定义提供重要的见解怀孕期间健康人类生殖组织中 DC 的功能特化，也提供了为将来研究先兆子痫等病理学奠定基础。