Complement Activation Signatures in Systemic Lupus Erythematosus: Castle Study

系统性红斑狼疮中的补体激活特征：Castle 研究

• 批准号: 9317177
• 负责人: John Atkinson
• 金额: $ 20.13万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-15 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9317177
• 关键词: Acute-Phase Proteins; Antibodies; Antigen-Antibody Complex; Apoptotic; Autoantibodies; Autoimmune Diseases; Automobile Driving; B-Lymphocytes; Binding; Biological Assay; Biological Markers; Blood; Blood Platelets; C3bi; Cell surface; Cells; Clinic; Clinical; Complement; Complement 3b; Complement 3d; Complement Activation; Complement Receptor; Complex; Consumption; Cytometry; Data; Deposition; Detection; Deterioration; Development; Devices; Diagnosis; Disease; Enrollment; Enzyme-Linked Immunosorbent Assay; Equilibrium; Erythrocytes; Flare; Flow Cytometry; Functional disorder; Gene Dosage; Generations; Glucocorticoids; Glucose; Goals; Half-Life; Heavy Metals; Heterogeneity; Human; Immune; Immune Cell Activation; Immune system; Immunologist; Individual; Inflammation; Inflammatory Response; Investigation; Isotopes; Lateral; Liver; Lupus; Lymphocyte; Mass Spectrum Analysis; Measurement; Measures; Medical Device; Membrane; Morbidity - disease rate; Organ; Pathway interactions; Patients; Phenotype; Pilot Projects; Production; Randomized Controlled Trials; Role; Serological; Serum; Surface; System; Systemic Lupus Erythematosus; Technology; Testing; Time; Tissues; Universities; Vitronectin; Washington; antibody conjugate; cell type; cohort; complement C3 precursor; complement C4d; complement system; design; diagnostic accuracy; ds-DNA; immune activation; improved; improved outcome; innovation; insight; mortality; pathogen; point of care; prospective; receptor; tool

Abstract Systemic lupus erythematosus (SLE) is an autoimmune disease that causes organ damage, leading to significant morbidity and mortality. SLE is characterized by the generation of autoantibodies, which bind to an individual’s own tissues to induce inflammation and organ damage. An important mechanism for SLE-induced inflammation is activation of the complement cascade. The complement system is an important part of the immune system, which is well-designed eliminate pathogens with the help of antibodies and other immune components. In SLE though, autoantibodies that have bound to patients’ organs activated complement to generate damaging inflammatory responses. Currently, clinicians assess complement activation through the complement components C3 and C4 in serum. As the complement cascade is activated, both C3 and C4 become attached to the surfaces of pathogens and cells and serum C3 and C4 levels decrease as a result. In SLE, since complement activation occurs during flares, C3 and C4 levels should also decrease in flares. During systemic inflammation though, the liver produces both C3 and C4. Thus, during SLE flares, there is both consumption (due to their activation on cell surfaces) and production (due to inflammation-induced liver production) of C3 and C4. This leads to the underdetection of SLE flares. Complement activation generates numerous split products (or fragments) that are found on the surface of cells and interact with numerous complement receptors (both stimulatory and regulatory). Given the central role of complement activation in SLE, identifying the array of complement signatures on immune cells and blood will likely generate important observations regarding SLE pathophysiology. Indeed, the value of complement split products have just started to be realized, as increased erythrocyte and platelet bound C4d can provide utility in the diagnosis of SLE. Nevertheless, a comprehensive, qualitative assessment of the complement fragment deposition on immune cells from patient with various states of SLE has yet to be performed. Here, we hypothesize identifying complement signatures on immune cells using mass cytometry and blood complement split products will provide invaluable insight in the role of complement activation on SLE. We have two aims to test this hypothesis: 1) Evaluate the complement split product iC3b as a dramatically improved biomarker of SLE disease activity, and 2) Fully delineate the complement signatures found on immune cells from patients with SLE. Two recent technological improvements have made evaluating these Aims possible: 1) Development of an investigational medical device that rapidly determines blood iC3b and C3 levels without artefactual elevation of iC3b; and 2) Highly multiplexed phenotyping tools for immune cells such as mass cytometry has emerged as an innovative approach to analyze complex multicellular systems. The device that measures iC3b and C3 does so within 20 minutes, and our preliminary data demonstrate a strong correlation between iC3b and C3 levels to SLE disease activity. Applying MC to the study of complement signatures on immune cells in SLE will provide a level of detail and quantification that has not been possible. Thus, rather than evaluating an incomplete profile of complement proteins, we have the potential to quantify the levels and types of complement fragments, complement receptors, and membrane regulators on multiple cell types at the single cell level. This proposal will help establish the role of iC3b and iC3b/C3 ratios as a promising approach to assessing SLE disease activity, and significantly improve our understanding of how cell surface complement activation signatures on immune cells drive pathophysiology in SLE.

摘要 系统性红斑狼疮（SLE）是一种自身免疫性疾病，可导致器官损伤， 严重的发病率和死亡率。SLE的特征在于自身抗体的产生，其结合至 这可能会损害个体自身的组织，从而诱发炎症和器官损伤。SLE诱导的一个重要机制 炎症是补体级联的激活。补体系统是免疫系统的重要组成部分， 免疫系统，这是精心设计的消除病原体的帮助下，抗体和其他免疫 件.然而，在SLE中，与患者器官结合的自身抗体激活了补体， 产生破坏性的炎症反应 目前，临床医生通过血清中的补体成分C3和C4评估补体激活。 当补体级联被激活时，C3和C4都附着在病原体的表面， 细胞和血清C3和C4水平因此降低。在SLE中，由于补体激活发生在 在耀斑中，C3和C4水平也应降低。在全身炎症期间，肝脏 产生C3和C4。因此，在SLE发作期间，存在两种消耗（由于它们在细胞上的激活）， 表面）和C3和C4的产生（由于炎症诱导的肝脏产生）。这导致 SLE发作的检测不足。 补体激活产生大量的分裂产物（或片段），这些产物存在于细胞表面 并与许多补体受体（刺激性和调节性）相互作用。由于核心作用， SLE中的补体激活，识别免疫细胞和血液上的补体特征阵列将 可能产生关于SLE病理生理学的重要观察结果。事实上，互补分裂的价值 产品刚刚开始实现，因为增加的红细胞和血小板结合的C4d可以提供实用性， SLE的诊断然而，对补体片段的全面定性评估 在来自具有各种SLE状态的患者的免疫细胞上的沉积还有待进行。 在这里，我们假设使用大量细胞计数和血液分析来识别免疫细胞上的补体特征。 补体裂解产物将为补体激活在SLE中的作用提供宝贵的见解。我们有 两个目的是检验这一假设：1）评估补体分裂产物iC 3b作为显著改善的 SLE疾病活动的生物标志物，和2）完全描绘免疫细胞上发现的补体特征 从SLE患者身上。 最近的两项技术改进使评估这些目标成为可能：1）开发一种 快速测定血液iC 3b和C3水平而不会人为升高 iC 3b;和2）用于免疫细胞的高度多重表型分析工具，如质谱细胞术，已经出现， 一种分析复杂多细胞系统的创新方法。测量iC 3b和C3的设备 所以在20分钟内，我们的初步数据表明iC 3b和C3水平之间存在很强的相关性， SLE疾病活动。将MC应用于SLE免疫细胞上补体标记的研究将提供 这是一个不可能的细节和量化水平。因此，与其评估不完整的 补体蛋白质谱，我们有可能量化补体片段的水平和类型， 补体受体和单细胞水平上的多种细胞类型上的膜调节剂。 这一建议将有助于确立iC 3b和iC 3b/C3比值作为评估SLE的一种有前途的方法的作用 疾病活动，并显着提高我们对细胞表面补体激活的理解 免疫细胞上的特征驱动SLE的病理生理学。