Influence of polysialic acid on leukocyte migration

聚唾液酸对白细胞迁移的影响

基本信息

批准号：
10170217
负责人：
NICHOLAS MILTON STAMATOS
金额：
$ 49.98万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-06-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10170217
关键词：
Adhesions Adhesives Adoptive Transfer Affect Bacterial Pneumonia Binding Binding Proteins Bone Marrow Bone Marrow Transplantation Carbohydrates Carrier Proteins Cell Adhesion Molecules Cell Maturation Cell Surface Proteins Cell Surface Receptors Cell physiology Cell surface Cells Clinical Coupled Dendritic Cells Discipline Disease E-Selectin Endothelial Cells Engineering Enzymes Flow Cytometry Genes Goals Health Homing Human Immune Immune Targeting Immune response Immune system Immunoprecipitation In Situ In Vitro Individual Infection Infectious Agent Inflammation Influenza A Virus, H1N1 Subtype Isotope Labeling Knockout Mice Leukocytes Ligand Binding Ligands Lipids Lung Lung infections Lymphatic Lymphatic System Mass Spectrum Analysis Modification Mus Myeloid Cells NCAM1 gene Neural Cell Adhesion Molecules Neuraxis Neurons Neuropilin-2 Neutrophil Infiltration Oncology Organ Outcome Patients Pattern Phagocytosis Physiological Processes Polysaccharides Polysialic Acid Positron-Emission Tomography Post-Translational Protein Processing Process Property Proteins Recruitment Activity Rheumatology Role Site Solid Streptococcus pneumoniae Surface Testing Viral Pneumonia Wild Type Mouse adaptive immune response axonal pathfinding cell motility chemokine design glycosylation improved in vivo influenzavirus interest macrophage magnetic beads member migration monocyte monolayer mouse model neutrophil novel strategies pathogen pathogenic bacteria pathogenic virus peripheral blood receptor recruit respiratory pathogen response vaccinology vascular bed

项目摘要

PROJECT SUMMARY Changes in the glycosylation pattern of cell surface proteins and lipids are increasingly being recognized as an important factor in controlling cellular activity. There is growing interest in how these changes can be regulated to affect physiologic processes in health and disease. The goal of this proposal is to understand how a specific carbohydrate modification on the surface of cells of the immune system influences the function of these cells during states of inflammation and infection. As a key member of cellular innate and adaptive immune responses, monocytes are recruited to pulmonary sites of infection where they differentiate into macrophages and dendritic cells before migrating through the lymphatic system. This targeted migration and programmed maturation of monocytes and monocyte-derived cells is guided by specific, highly-organized cell-cell and receptor-ligand interactions, many of which are modulated by glycosylation of cell surface receptors and adhesion molecules. Polysialic acid (polySia) is a unique glycan modification of at least three such proteins, neural cell adhesion molecule (NCAM/CD56), neuropilin-2 (NRP-2), and E-selectin ligand-1 (ESL-1), that are expressed at different stages of monocyte maturation. We will test the hypothesis that regulated expression of polySia on monocytes as they differentiate into macrophages and dendritic cells, and on neutrophils that also are polysialylated, helps direct cell homing and a well-orchestrated immune response during pulmonary infection with viral and bacterial pathogens. Mice that are deficient in expression of the enzyme that synthesizes polySia in leukocytes (ST8 SiaIV-/-) and of the carrier proteins (NCAM/CD56-/- and NRP-2-/-) will be used for in vitro and in vivo studies. Our Specific Aims will i) establish the impact in vivo of polySia on monocytes, macrophages, DC and neutrophils in the targeted immune response in a murine model of bacterial and viral pneumonia; ii) define the mechanism(s) through which polysialylated proteins on the surface of human and murine monocytes, macrophages, DC and neutrophils control adhesion to and migration across pulmonary microvascular monolayers; and iii) define and analyze the regulated expression of polySia and of polysialylated proteins during differentiation of primary monocytes into DC and macrophages. We expect to identify specific cell-cell and receptor-ligand interactions that polySia promotes or interferes with during different stages of the monocyte/macrophage/dendritic cell maturation and migratory processes, as well during neutrophil recruitment. The results from our studies will provide a blueprint to engineer levels of expression of polySia and/or carrier proteins in myeloid cells in order to optimize migration to and from sites of infection/inflammation to improve the overall immune response.

项目摘要细胞表面蛋白和脂质的糖基化模式的变化越来越被认为是一种控制细胞活性的重要因素。人们对如何调节这些变化越来越兴趣影响健康和疾病的生理过程。该提议的目的是了解特定的免疫系统细胞表面上的碳水化合物修饰会影响这些细胞的功能在炎症和感染状态。作为细胞先天和适应性免疫的关键成员反应，单核细胞募集到感染的肺部，它们分化为巨噬细胞在通过淋巴系统迁移之前，树突状细胞。这个有针对性的迁移和编程单核细胞和单核细胞衍生细胞的成熟由特定的，高度组织的细胞和受体配体相互作用，其中许多是通过细胞表面受体糖基化和粘附分子。多氨酸（polysia）是至少三种此类蛋白质的独特的聚糖修饰，神经细胞粘附分子（NCAM/CD56），Neuropilin-2（NRP-2）和E-选择蛋白配体1（ESL-1），是在单核细胞成熟的不同阶段表达。我们将检验以下假设单核细胞上的polysia分化为巨噬细胞和树突状细胞，以及嗜中性粒细胞被多磷酸化，有助于导致细胞归巢和肺部良好的免疫反应病毒和细菌病原体感染。缺乏表达酶的小鼠合成白细胞（ST8 SIAIV - / - ）和载体蛋白（NCAM/CD56 - / - 和NRP-2 - / - ）中的polysia 用于体外和体内研究。我们的具体目的是）i）建立Polysia的体内影响在细菌的鼠模型中，靶向免疫反应中的单核细胞，巨噬细胞，DC和中性粒细胞和病毒肺炎； ii）定义了在表面上多裂蛋白的机制人和鼠的单核细胞，巨噬细胞，DC和中性粒细胞控制着对遍布和迁移的附着肺微血管单层； iii）定义和分析了多利亚和的调节表达在原代单核细胞分化为直流和巨噬细胞的过程中，多磷酸蛋白质。我们希望确定polysia促进或干扰期间的特定细胞和受体配体相互作用单核细胞/巨噬细胞/树突状细胞的成熟和迁移过程的不同阶段以及在中性粒细胞招募。我们研究的结果将为工程师的表达水平提供蓝图髓样细胞中的polysia和/或载体蛋白，以优化迁移至感染/炎症以改善总体免疫反应。