Physiological vs. pharmacological effects of glucocorticoids on human monocytes.

糖皮质激素对人单核细胞的生理与药理作用。

• 批准号: 8114343
• 负责人: PAUL GUYRE
• 金额: $ 27.65万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-15 至 2013-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8114343
• 关键词: Acute; Address; Adrenal hormone preparation; Anti-Inflammatory Agents; Anti-inflammatory; Autoimmunity; Behavioral Research; Biological Assay; Biomedical Research; Cells; Clinical; Clinical Protocols; Clinical Research; Data; Development; Dose; Effectiveness; Enzyme-Linked Immunosorbent Assay; Escherichia coli; Evaluation; Event; Exposure to; Flow Cytometry; Fostering; Future; Genes; Glucocorticoids; Human; Hydrocortisone; Immune; Immune response; Immunity; In Vitro; Individual; Individual Differences; Infection; Inflammation; Inflammatory; Inflammatory Response; Interleukin-6; Lead; Leukocytes; Lipopolysaccharides; Measurement; Mediator of activation protein; Messenger RNA; Mitogen-Activated Protein Kinases; Molecular; Mononuclear; Pathway interactions; Phosphorylation; Physiological; Pilot Projects; Play; Production; Regulation; Reporting; Role; Saline; Screening procedure; Steroids; Stress; Testing; Time; Trauma; chemokine receptor; clinically significant; cytokine; design; exposed human population; human study; immune activation; improved; in vivo; innovation; macrophage; monocyte; novel; novel therapeutic intervention; peripheral blood; response

DESCRIPTION (provided by applicant): Pharmacological glucocorticoids (GCs) have been effectively used to treat inflammation for over a half century, resulting in the established paradigm of GC action that addresses only mechanisms that lead to suppression of inflammation and immunity. In contrast, and less appreciated, is the fact that physiological stress-associated concentrations of GCs-but not higher pharmacological levels-play an essential enhancing role in the activation of immune and inflammatory responses. We therefore propose a pilot human study to obtain data that will support a new paradigm, one addressing the mechanisms by which a transient in vivo increase in cortisol to stress-associated levels (e.g., levels that follow trauma) prepares for an enhanced innate immune response when a subsequent activation event occurs (e.g., an infection). Our study is designed to test the hypothesis that preparative phenotypic and molecular changes induced by physiological concentrations of cortisol in human monocytes and macrophages, lead to functionally opposing consequences compared with pharmacological GC doses. Our secondary aim, identifying mechanisms for individual-specific differential responses to 'stress- cortisol' versus 'pharm-cortisol' would, if successful, provide a screening approach with far reaching benefits for nearly all avenues of biomedical, behavioral, and clinical research. Specifically, we propose to: 1. Identify specific cellular and molecular changes that distinguish stress-cortisol enhancement from pharm- cortisol suppression of human monocyte/macrophage innate immune activation pathways. We have found that stress-cortisol pretreatment reproducibly enhances LPS-induced IL-6 production by human monocyte-derived macrophages from some subjects (responders) but not others (non-responders). Inter-individual differences and differential effects of stress- vs. pharm-cortisol will be used to discriminate by flow cytometry, gene profiling analysis and ELISA assays the most relevant molecular events that lead to stress-cortisol enhanced cytokine production. We will specifically interrogate mechanisms by which stress-cortisol enhancement of LPS- induced MAP kinase phosphorylation augments innate immune activation in human macrophages. 2. Identify, at the molecular level, specific monocyte responses that are differentially regulated by stress- cortisol and pharm-cortisol in vivo. Each subject will serve as his/her own control, and will receive 3 in vivo treatments; saline, stress-cortisol (40mg/70kg/6hours), or pharm-cortisol (400mg/70kg/6hours) with an inter- treatment interval that returns measurements to baseline. Flow cytometry, Taqman real time PCR and ELISA assays will be used to determine the in vivo onset, magnitude and durability of stress-cortisol versus pharm- cortisol-induced changes in key cellular regulators of innate immune inflammation. We plan to identify specific molecular events that predict differential in vivo effects of stress-cortisol versus pharm-cortisol, or differences in innate immune activation among individual subjects. Our results will direct future studies to determine important clinical distinctions between anti-inflammatory and pro-inflammatory regulatory effects of GCs. PUBLIC HEALTH RELEVANCE: Steroids related to the natural adrenal hormone hydrocortisone have been used to effectively treat inflammation and autoimmunity for over a half century, but unexplained differences in their effectiveness in different people presents a vexing clinical challenge. Improved understanding of the mechanisms by which these steroids regulate mediators of immunity and inflammation is needed to foster new therapeutic interventions for the treatment of acute systemic inflammation. The studies proposed are novel in that, to the best of our knowledge, we will be the first to examine mechanisms by which physiological stress-associated levels of hydrocortisone enhance immune and inflammatory pathways within human monocytic white blood cells in vivo.

描述（由申请人提供）：半个多世纪以来，药理糖皮质激素（GCs）已被有效地用于治疗炎症，导致GC作用的建立范例仅解决导致炎症和免疫抑制的机制。与此相反，很少有人认识到的是，生理应激相关的gcs浓度（而不是更高的药理学水平）在免疫和炎症反应的激活中起着重要的增强作用。因此，我们建议进行一项人体初步研究，以获得支持新范式的数据，该范式解决了皮质醇在体内短暂增加到与压力相关的水平（例如，创伤后的水平）的机制，该机制为随后的激活事件（例如感染）发生时先天免疫反应的增强做准备。我们的研究旨在验证这样一种假设，即与GC药理学剂量相比，人单核细胞和巨噬细胞中皮质醇的生理浓度引起的制备性表型和分子变化会导致功能上相反的后果。我们的第二个目标是确定个体对“压力-皮质醇”和“药物-皮质醇”的不同反应机制，如果成功的话，将为生物医学、行为和临床研究的几乎所有途径提供一种具有深远益处的筛选方法。具体而言，我们建议：1。鉴定特异性细胞和分子变化，区分应激皮质醇增强和药物皮质醇抑制人类单核细胞/巨噬细胞先天免疫激活途径。我们发现应激皮质醇预处理可重复性地增强lps诱导的人单核细胞源性巨噬细胞产生IL-6，这些巨噬细胞来自一些受试者（应答者），而不是其他受试者（无应答者）。个体间差异和应激与药物皮质醇的差异效应将通过流式细胞术、基因谱分析和ELISA分析来区分导致应激皮质醇增强细胞因子产生的最相关分子事件。我们将特别探究LPS诱导的应激皮质醇增强MAP激酶磷酸化增强人巨噬细胞先天免疫激活的机制。2. 鉴定，在分子水平上，特异性单核细胞反应是由应激皮质醇和药物皮质醇在体内的差异调节。每位受试者作为自己的对照，接受3次体内治疗；生理盐水，应激皮质醇（40mg/70kg/6小时），或药物皮质醇（400mg/70kg/6小时），治疗间隔使测量值恢复到基线。流式细胞术、Taqman实时PCR和ELISA检测将用于确定体内应激皮质醇与药物皮质醇诱导的先天免疫炎症关键细胞调节因子变化的发生、程度和持久性。我们计划确定特定的分子事件，预测应激皮质醇与药物皮质醇在体内的差异效应，或个体受试者先天免疫激活的差异。我们的结果将指导未来的研究，以确定GCs的抗炎和促炎调节作用之间的重要临床区别。