Cellular Sources of Self Antigen in SLE

SLE 自身抗原的细胞来源

基本信息

批准号：
10265341
负责人：
DAVID STEPHEN PISETSKY
金额：
--
依托单位：
DURHAM VA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-01-01 至 2022-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10265341
关键词：
Acetaminophen Address Affect Alpha Particles Anti-DNA Antibodies Antibodies Antibody Specificity Antigen-Antibody Complex Antinuclear Antibodies Apoptosis Appearance Attention Autoantigens Autoimmune Diseases Autoimmunity Bacteria Binding Biochemical Biological Assay Biological Markers Blood Cell Death Cell Line Cells Cessation of life Clinical Research Complex Complication Confocal Microscopy DNA Data Dendritic Cells Deposition Development Diagnosis Disease Electrons Extracellular Space Flow Cytometry Galactosamine Generations Guanosine Health Immune Immune response Immune system Immunologics Immunosuppressive Agents Impairment In Vitro Infection Inflammation Inflammatory Interferons Kidney Lupus Lymphoid Cell Malignant Neoplasms Mediating Membrane Metabolism Mitochondria Mitochondrial DNA Molecular Monoclonal Antibodies Mus Myeloid Cells Nature Nephritis Nuclear Outcome Pain Particulate Pathogenesis Pathogenicity Patients Production Prognosis Property Quality of life Research Scanning Electron Microscopy Serology Source Structure System Systemic Lupus Erythematosus Techniques Tissues Toxic effect Uncertainty Veterans Woman Work cell injury cellular imaging cytokine disability experimental study extracellular in vivo in vivo Model insight macromolecule military veteran mouse model new therapeutic target novel marker novel strategies oxidation particle sensor vesicular release

项目摘要

Abstract Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease characterized by the production of antinuclear antibodies (ANAs) in association with severe multisystem inflammatory disease manifestations. ANAs target a wide array of nuclear macromolecules and can mediate disease by the formation of immune complexes that deposit in the tissue to promote inflammation and damage; these complexes can also stimulate the production of cytokines including type 1 interferon to drive generalized immune system disturbances. Among ANAs forming immune complexes, antibodies to DNA (anti-DNA) are the serological hallmark of SLE and markers of diagnosis and prognosis. These antibodies bind to both single and double stranded DNA and form immune complexes that deposit in the tissue, especially the kidney, to promote inflammation; in addition, these immune complexes can stimulate the production of cytokines by plasmacytoid dendritic cells, triggering internal sensors of DNA. While the properties of anti-DNA antibodies have been extensively studied, much less is known about the origin of antigenic DNA, its access to the immune system and its unique molecular properties. In general, the source of this DNA has been considered to be nuclear in origin and the product of dead and dying cells. Our studies have provided a new perspective on this issue by demonstrating two important features of extracellular DNA: 1) its existence in the form of microparticles and 2) the presence in these particles of mitochondrial (mt) as well as nuclear DNA (nDNA). This presence of mtDNA is important since mtDNA is intrinsically immunostimulatory because of structural features that differ from those of nDNA. These involve CpG motifs in mtDNA, a consequence of the origin of mitochondria from bacteria, and oxidation of guanosine residues. Furthermore, we have shown that mitochondria released from cells have properties similar to those of microparticles, suggesting that mitochondria may serve as self-antigens in lupus, contributing to immune activities attributed to microparticles and providing a nidus for immune complex formation. Building on preliminary work, the proposed studies will focus on three main hypotheses: 1) DNA autoantigen can be released from cells undergoing various death forms and exist in both a free and particle form; 2) the metabolism of DNA during cell death influences the amount in the blood, its size and its representation in particle or soluble form; and 3) anti-DNA antibodies can bind to various antigenic forms of DNA, both free and particulate, with assay of antibodies to these forms providing more informative biomarkers. We will pursue three specific aims. Specific Aim 1: To determine the extracellular release of cellular DNA during different forms of in vitro cell death. We will induce different forms of cell death in cell lines in vitro and determine the representation of mtDNA and nDNA in soluble and particulate forms; Specific Aim 2. To use in vivo models to elucidate the release of DNA into the blood. We will use in vivo systems in the mouse to explore the release of mtDNA and nDNA following cell death; we will also sequence DNA in the blood by high throughput techniques; and Specific Aim 3: Using blood from patients with lupus and murine models, we will determine the presence and specificity of antibodies to various sources of extracellular DNA in SLE. To develop new serological approaches, we will determine the presence and specificity of antibodies to different antigenic forms of DNA, both free and particulate and, by assessing results from a large panel of well characterized patients, develop new biomarkers. Successful completion of these experiments will provide new insights into the mechanisms of pathogenicity as well as provide new biomarkers for clinical and research purposes.

抽象的全身性红斑狼疮（SLE）是一种原型自身免疫性疾病，其特征是产生抗核抗体（ANA）与严重的多系统炎症性疾病表现相关。 ANA靶向各种各样的核大分子，可以通过免疫形成介导疾病沉积在组织中以促进炎症和损害的复合物；这些复合物也可以刺激包括1型干扰素在内的细胞因子的产生，以驱动广义免疫系统干扰。在形成免疫复合物的ANA中，DNA的抗体（抗DNA）是SLE的血清学标志以及诊断和预后的标记。这些抗体与单链DNA和双链DNA结合，形成沉积在组织中的免疫复合物，尤其是肾脏，以促进炎症；此外，这些免疫复合物可以刺激细胞晶状体树突状细胞的细胞因子产生，从而触发 DNA的内部传感器。虽然已经对抗DNA抗体的特性进行了广泛的研究，但很多关于抗原DNA的起源，其访问免疫系统及其独特的分子的了解鲜为人知特性。通常，该DNA的来源被认为是起源的核和死亡和垂死的细胞。我们的研究通过证明两个细胞外DNA的重要特征：1）其以微粒的形式存在，2）这些线粒体（MT）和核DNA（NDNA）的颗粒。 mtDNA的存在很重要由于mtDNA具有与NDNA不同的结构特征，因此具有本质上的免疫刺激性。这些涉及mtDNA中的CpG基序，这是细菌线粒体的起源和氧化的结果鸟嘌呤残留物。此外，我们已经表明，从细胞释放的线粒体具有特性与微粒类似，表明线粒体可以用作狼疮的自我抗原，有助于归因于微粒的免疫活性，并为免疫复合物提供Nidus 形成。在初步工作的基础上，拟议的研究将重点放在三个主要假设上：1）DNA 自动抗原可以从经历各种死亡形式的细胞中释放出来，并且存在于自由和粒子中形式; 2）细胞死亡期间DNA的代谢影响血液，大小及其的数量以粒子或可溶性形式表示； 3）抗DNA抗体可以与各种抗原形式结合自由和颗粒物的DNA，对这些形式的抗体进行测定，提供了更有信息的生物标志物。我们将追求三个具体目标。特定目标1：确定细胞外DNA的细胞外释放在不同形式的体外细胞死亡中。我们将在体外诱导细胞系中不同形式的细胞死亡，并且在可溶性和颗粒形式中确定mtDNA和nDNA的表示；特定目标2。体内模型以阐明DNA释放到血液中。我们将在鼠标中使用体内系统探索细胞死亡后mtDNA和nDNA的释放；我们还将通过高的血液对DNA进行序列DNA 吞吐技术；和特定的目标3：使用狼疮和鼠模型患者的血液，我们将确定SLE中各种细胞外DNA来源的抗体的存在和特异性。到开发新的血清学方法，我们将确定抗体对不同的抗体的存在和特异性抗原形式的DNA，无论表征患者，发展新的生物标志物。这些实验的成功完成将提供新的洞悉致病性机制，并为临床和研究提供新的生物标志物目的。