Determinants of human neutrophil fate after phagocytosis

吞噬作用后人类中性粒细胞命运的决定因素

• 批准号: 10092904
• 负责人: William M. Nauseef
• 金额: $ 48.68万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-02-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10092904
• 关键词: Apoptosis; Apoptotic; Cell Death; Cells; Cessation of life; Complex; Cytoplasmic Granules; Disease; Environment; Failure; Generations; Health; Homeostasis; Host Defense; Human; Infection; Inflammasome; Inflammation; Inflammatory; Inflammatory Response; Ingestion; Innate Immune Response; Interleukin-1 beta; Invaded; Membrane; Microbe; Mission; NADPH Oxidase; Necrosis; Neisseria gonorrhoeae; Oxidants; Pathway interactions; Phagocytes; Phagocytosis; Phagosomes; Phase; Phosphotransferases; Production; Proliferating Cell Nuclear Antigen; Protein Kinase; Proteins; RIPK1 gene; Resolution; Role; Serine Protease; Signal Pathway; Signal Transduction; Staphylococcus aureus; System; Time; Tissues; cytokine; first responder; human pathogen; insight; macrophage; microbicide; neutrophil; novel; receptor; tool

Human innate immune response to microbes depends on the coordinated interactions among a variety of cells and secreted factors, with polymorphonuclear leukocytes (PMN) typically prominent among the first responders. In their capacity as phagocytic cells, PMN sequester ingested prey in membrane-bound phagosomes, where oxidants from the NADPH oxidase and proteins from granules synergize to create a toxic environment that promotes death and degradation of the engulfed microbe. In certain settings, a significant fraction of ingested microbes survives within PMN. The persistence of viable microbes within phagosomes not only provides a mechanism for sustained infection but also can modulate the local inflammatory tone by altering the programmed cell death of PMN and promoting PMN release of proinflammatory cytokines. Interference with phagocytosis-induced apoptosis of PMN (PICD), either by delaying apoptosis, as seen with N. gonorrhoeae (Ngc) or engaging a novel necrotic cell death pathway, as seen with Staphylococcus aureus (SA), thwarts resolution of the inflammatory response and causes release of host-derived danger signals that promote inflammation and secondary tissue damage. Thus, effective innate immune response requires not only death and degradation of invading microbes but also resolution of inflammation and reestablishment of homeostasis. The overarching hypothesis of this proposal is that the failure of PMN to undergo apoptotic cell death derails the resolution phase of the inflammatory response and instead amplifies disease. Because activated human PMN (hPMN) and their secreted products can sculpt the inflammatory tone in tissue, we propose to use Ngc and SA -- both human pathogens that survive within PMN, alter phagocytosis- induced cell fate pathways and elicit profound inflammatory local changes – as tools to probe mechanisms that dictate phagocyte fate and timely resolution of inflammation. We will pursue two Specific Aims: Aim 1: To determine the mechanisms that regulate human PMN fate . A. Determine the signaling pathways that differentially direct hPMN towards survival vs programmed cell death (apoptosis vs primary necrosis). B. Determine the composition and activities of the ripoptosome (intracellular complexes) in hPMN after phagocytosis C. Determine the role of Proliferating Cell Nuclear Antigen (PCNA) in the fate of phagocytosing hPMN Aim 2: To determine the mechanisms underlying hPMN secretion of IL-1β during phagocytosis A. Identify the role of inflammasomes in IL-1β production by hPMN during phagocytosis B. Determine the importance of serine proteases in generation of IL-1β C. Determine the role of Receptor-interacting kinase-3 (RIPK-3) in hPMN IL-1β secretion

人类对微生物的先天免疫反应取决于各种各样的协调相互作用 细胞和分泌因子，多形核白细胞（PMN）通常在第一个中突出 响应者。 PMN以其作为吞噬细胞的能力，在结合膜结合的猎物中摄取猎物 吞噬体，其中来自氧化物NADPH氧化物的氧化剂和颗粒的蛋白质协同作用以产生有毒 促进吞噬微生物的死亡和退化的环境。在某些情况下，重要的 摄入的微生物的一部分在PMN内生存。吞噬体中可行微生物的持久性而不是 仅提供持续感染的机制，但也可以通过 改变PMN的程序性细胞死亡并促进促炎细胞因子的PMN释放。 干扰吞噬作用引起的PMN凋亡（PICD），要么通过延迟凋亡，如所见 N. Gonorrhoeae（NGC）或与金黄色葡萄球菌一起观察到的新型坏死细胞死亡途径 （sa），阻止炎症反应的解决，并导致释放宿主衍生的危险信号 促进炎症和继发组织损伤。那是有效的先天免疫反应不需要 仅死亡和入侵微生物的死亡和降解，也只能解决炎症和重新建立 稳态。该提议的总体假设是，PMN未能经历凋亡 细胞死亡使炎症反应的分辨率阶段和放大器疾病的分辨阶段。 因为被激活的人PMN（HPMN）及其分泌的产品可以雕刻 组织，我们建议使用NGC和SA-这两种人的病原体都在PMN中生存，改变了吞噬作用 - 诱导的细胞命运途径并引起深刻的炎症局部变化 - 作为探测机制的工具 决定吞噬细胞的命运，并及时解决炎症。我们将追求两个具体的目标： 目标1：确定调节人PMN命运的机制 。 答：确定不同将HPMN转向生存与编程的信号通路 细胞死亡（凋亡与原发性坏死）。 B.确定HPMN中ripopoptosom（细胞内复合物）的组成和活性 吞噬作用 C.确定增殖细胞核抗原（PCNA）在吞噬HPMN的命运中的作用 目标2：确定吞噬作用期间IL-1β的HPMN分泌的机制 A.确定炎症在吞噬作用过程中HPMN产生IL-1β在IL-1β中的作用 B.确定丝氨酸蛋白在IL-1β产生中的重要性 C.确定受体相互作用激酶3（RIPK-3）在HPMN IL-1β分泌中的作用