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.

抽象的 全身性红斑狼疮（SLE）是一种自身免疫性疾病，会造成器官损伤，导致 显着的发病率和死亡率。 SLE的特征是自动抗体的产生，该自身抗体结合到一个 个人自身的组织会诱导注射和器官损伤。 SLE引起的重要机制 炎症是完成级联的激活。完成系统是 免疫系统，精心设计可消除病原体，借助于抗体和其他免疫 成分。但是，在SLE中，与患者器官绑定的自身抗体激活完成 产生破坏性炎症反应。 目前，临床医生通过完成血清中的C3和C4评估完成激活。 随着完成级联激活的激活，C3和C4都附着在病原体的表面和 细胞和血清C3和C4水平结果降低。在SLE中，由于完成激活在 耀斑，C3和C4水平也应降低耀斑。但是，在系统性炎症期间，肝脏 同时产生C3和C4。在SLE耀斑期间，这两者都有消耗（由于它们在细胞上的激活 表面）和C3和C4的生产（由于炎症引起的肝脏产生）。这导致了 SLE耀斑的检测不足。 补体激活会产生许多在细胞表面发现的分裂产品（或片段） 并与许多补体受体（刺激和调节性）相互作用。鉴于中心作用 SLE中的补体激活，识别免疫细胞和血液上的补体特征会 可能会产生有关SLE病理生理学的重要观察。确实，完成的价值分配 由于红细胞和血小板绑定的C4D可以提供实用程序，因此产品刚刚开始实现 SLE的诊断。然而，对完成片段的全面，定性评估 尚未进行各种SLE状态的患者对免疫细胞的沉积。 在这里，我们假设使用质量细胞仪和血液鉴定免疫细胞上的补体特征 补体拆分产品将为SLE的完成激活作用提供宝贵的见解。我们有 两个目的是检验以下假设：1）评估拆分产品IC3B作为大幅改进 SLE疾病活性的生物标志物，以及2）完全描述免疫细胞上的补体特征 来自SLE患者。 最近的两项技术改进使评估这些目的成为可能：1）开发 迅速确定血液IC3B和C3水平的研究性医疗设备，而没有人工升高 IC3B; 2）用于免疫细胞（例如质量细胞术）的高度多重表型工具已出现为 一种创新的方法来分析复杂的多细胞系统。测量IC3B和C3的设备确实 因此，在20分钟内，我们的初步数据证明了IC3B和C3水平与 SLE病活动。将MC应用于SLE的免疫细胞的完成签名的研究将提供 不可能的细节和数量水平。那，而不是评估不完整的 补体蛋白质的特征，我们有可能量化补体片段的水平和类型， 在单细胞水平的多种细胞类型上的补体受体和膜调节剂。 该建议将有助于确定IC3B和IC3B/C3比率作为评估SLE的承诺方法 疾病活动，并显着提高我们对细胞表面完成如何激活的理解 免疫细胞的签名驱动SLE的病理生理学。