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：
10401230
负责人：
Martin Prlic
金额：
$ 6.39万
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 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10401230
关键词：
Activities of Daily Living Agonist Allogenic Antigen Presentation Antigen-Presenting Cells Antigens Autoantigens Autoimmunity Biological Assay Biological Markers Blood Cells Cellular Indexing of Transcriptomes and Epitopes by Sequencing 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 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 reproductive outcome 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.

孕产妇免疫适应怀孕需要并发维持免疫能力半合同性胎儿的特定耐受性。免疫适应与几个有关不良怀孕结果，对孕产妇和后代健康的短期和长期影响。孕产妇在怀孕期间暴露于胎儿的抗原；但是，很多问题仍然存在抗原表现和由此产生的适应性免疫反应的性质。引起从头自适应免疫反应需要专业抗原呈递细胞（APC）的激活，特别是树突状细胞（DC）。重要的是，DC还可以诱导T细胞的耐受性，从而在避免不必要的免疫反应中起关键作用。这通常是在防止对自我抗原的反应的背景下进行的，并且与保护胎儿免受母体排斥相关。人类胎盘和DECIDUA中的APC如何提供免受感染的保护，同时又可以防止对胎儿的免疫反应。了解人类繁殖的保护和耐受性的主要障碍是有关这些APC的功能概况及其个体间变化的有限信息。我们建议在母体血液，胎盘，Decidua和脐带血液中定义人类APC亚群，以检验以下假设：与人类繁殖有关的APC由几个在功能上专门用于保护或耐受性的子集组成。为此，我们将利用单细胞分析领域的最新技术进步，即高度高维数细胞术和单细胞基因表达分析，以剖析功能异质性这种免疫室。我们的实验室已经开发并验证了多个28色荧光细胞仪测定询问细胞表型，激活和功能能力的测定（即精疲力尽的表达标记，抑制受体，共刺激分子）。我们将使用这些测定法来确定细胞直接离体的组成和能力。这些实验将提供定义的关键见解在怀孕期间，健康人类生殖组织中DC的功能专业化，也提供研究未来诸如先兆子痫等病理的基础。