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: 8227989
• 负责人: PAUL GUYRE
• 金额: $ 15.8万
• 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 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8227989
• 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; Health; 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）：有效地使用药理学糖皮质激素（GC）来治疗炎症，导致了GC作用的已建立范式，该范例仅解决导致抑制炎症和免疫力的机制。相比之下，较不受欢迎的事实是，生理压力相关的GCS浓度，但不是更高的药理学水平 - 扮演在免疫和炎症反应激活中的基本增强作用。因此，我们提出了一项试验人类研究，以获取支持新范式的数据，该数据解决了一种机制，该机制通过这种机制，使皮质醇的瞬时在体内增加到压力相关水平（例如，遵循创伤的水平）为增强的先天免疫反应做好准备时，随后的激活事件发生时（例如，感染）。我们的研究旨在检验以下假设：人类单核细胞和巨噬细胞中皮质醇的生理浓度诱导的制剂表型和分子变化，与药理GC剂量相比会导致功能相反的后果。我们的次要目的是确定对“压力皮质醇”与“ Pharm-Cortisol”的个体特异性差异反应的机制，如果成功，将提供一种筛查方法，并为几乎所有生物医学，行为和临床研究的途径提供了遥远的益处。具体而言，我们提出：1。确定特定的细胞和分子变化，以区分应激皮质醇的增强与人类单核细胞/巨噬细胞先天性免疫激活途径的抑制性皮质醇。我们已经发现，应力 - 皮质醇预处理可重复地增强了来自某些受试者（反应者）的人类单核细胞衍生的巨噬细胞的LPS诱导的IL-6产生，但没有其他受试者（非反应者）。胁迫与皮质醇的个体间差异和差异作用将通过流式细胞仪，基因分析分析和ELISA分析来区分最相关的分子事件，从而导致胁迫皮质醇增强的细胞因子产生。我们将特别审问机制，通过这种机制，通过这种机制，应力 - 皮质醇增强LPS诱导的MAP激酶磷酸化增强了人类巨噬细胞中的先天免疫激活。 2。在分子水平上确定特定的单核细胞反应，这些反应受到胁迫 - 皮质醇和Pharm-Cortisol in Vivo的差异调节。每个主题都将作为自己的控制，并将获得3种体内治疗；盐水，应力 - 皮质醇（40mg/70kg/6小时）或Pharm-Cortisol（400mg/70kg/6小时），其间隔间隔将测量结果返回基线。流式细胞仪，TAQMAN实时PCR和ELISA测定法将用于确定应激皮质醇与药物 - 皮质醇诱导的固有免疫免疫炎症的关键细胞调节剂的变化的体内发作，大小和耐用性。我们计划确定特定的分子事件，以预测应激皮质醇与药物 - 皮质醇的体内差异差异，或者单个受试者之间先天免疫激活的差异。我们的结果将指导将来的研究确定GC的抗炎和促炎调节作用之间的重要临床区别。 公共卫生相关性：与天然肾上腺激素氢化可的松有关的类固醇已被用来有效治疗半个世纪以来的炎症和自身免疫性，但在不同人的有效性上无法解释的差异却带来了令人沮丧的临床挑战。对这些类固醇调节免疫和炎症调节介质的机制的理解有了改进的理解，以促进新的治疗干预措施以治疗急性全身炎症。提出的研究是新颖的，据我们所知，我们将是第一个研究生理胁迫相关水平的氢化可的松水平的机制，可以增强体内人单核细胞白细胞内的免疫和炎症途径。