Immune Suppression of Collagen Arthritis

胶原关节炎的免疫抑制

• 批准号: 7654937
• 负责人: Linda K. Myers
• 金额: $ 32.51万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7654937
• 关键词: Affinity; Alleles; Amino Acid Substitution; Amino Acids; Animal Model; Antibodies; Antigens; Arthritis; Arthroplasty; Autoimmune Process; Autoimmunity; Baculovirus Expression System; Binding; Biological Assay; CD3 Antigens; CD4 Positive T Lymphocytes; CD44 gene; Cartilage; Cell Count; Cell physiology; Cells; Chronic; Collagen; Collagen Arthritis; Collagen Type I; Collagen Type II; Competitive Binding; Complex; Cytolysis; DBA/1 Mouse; Data; Development; Disease; Down-Regulation; Electrophoretic Mobility Shift Assay; Enzyme-Linked Immunosorbent Assay; Epitopes; Event; Exposure to; Flow Cytometry; Foxes; Haplotypes; Harvest; Heart; Hybridomas; Hydroxylation; IL2RA gene; Immune; Immune response; Immunity; Immunization; Immunodominant Epitopes; Immunoprecipitation; Immunosuppressive Agents; Immunotherapy; In Vitro; Incubated; Inflammatory; Injury; Inositol Metabolism Pathway; Interleukin-17; Interleukin-4; JUN gene; Joints; Kinetics; Ligands; MAPK14 gene; MAPK8 gene; Major Histocompatibility Complex; Measures; Mediating; Microfluidics; Mitogen-Activated Protein Kinases; Modeling; Molecular; Mouse Strains; Mus; Nuclear; Oral Administration; Pathogenesis; Pathway interactions; Patients; Pattern; Peptides; Phenotype; Phosphatidylinositols; Phospho-Specific Antibodies; Phosphorylation; Physiologic pulse; Piceatannol; Pichia; Play; Population; Positioning Attribute; Post-Translational Protein Processing; Protein Tyrosine Kinase; Proteins; Protocols documentation; Receptor Signaling; Recombinants; Relative (related person); Rheumatoid Arthritis; Role; Serum; Severities; Signal Pathway; Signal Transduction; Site; Small Interfering RNA; Specimen; Spleen; Splenocyte; Staining method; Stains; Sulfonamides; System; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Subsets; TFRC gene; Testing; Time; Transgenes; Transgenic Mice; Transgenic Organisms; Ursidae Family; Western Blotting; ZAP-70 Gene; analog; autoimmune arthritis; base; c-myc Genes; cell type; cytokine; glycosylation; human SYK protein; inhibitor/antagonist; insight; lymph nodes; open label; peptide analog; prevent; public health relevance; research study; response; transcription factor; tripolyphosphate

DESCRIPTION (provided by applicant): Rheumatoid arthritis is a chronic inflammatory disease of diarthrodial joints. In preliminary studies we find that 50% of RA patients produce significant T cell responses to type II collagen (CII). Our hypothesis, that an antigen-driven autoimmune process mediates articular injury, is based on our findings that the oral administration of CII to arthritis patients in our open label trials resulted in favorable alterations in the cytokine profile measured. These data support the view that autoimmunity to an antigen(s) such as CII in cartilage plays a major role in the pathogenesis of the disease. We have used humanized mice bearing DRB1*0101 and DRB1*0401 transgenes and the collagen- induced arthritis (CIA) model to develop a collagen-based immunotherapy. By using proliferation and cytokine assays, we found that the core of the immunodominant determinant presented to murine T-cells by both DR1 and DR4 in Tg mice is CII263-270 (FKGEQGPK). Subsequently, a synthetic analog peptide was developed CII 263-273 (F263N, E266D) (A12) that was found to induce a profound suppression of CIA when administered after autoimmune arthritis has been established. Based on these data, we propose the following hypothesis: Interaction of the A12 analog peptide/APC complex with TCR leads to a unique signaling pathway, very likely involving phosphorylation of Syk rather than ZAP 70. The resulting cascade of signaling events induces predominantly Th2 cytokines and ultimately suppression of arthritis. We propose the following aims: 1) Identify T-cell signaling pathways activated by A12 and compare them with pathways induced by the wild type CII-peptide. 2) Characterize the inhibitory T cells induced by A12 and ascertain how this selected population can inhibit immunity to CII and autoimmune arthritis. 3) Determine the role of post-translational modifications in enhancing the efficacy of the A12 analog in suppressing immunity to CII and arthritis. These studies will determine what intracellular signaling pathways are triggered by A12, how these differ from those triggered by the unaltered peptide ligand and intact CII, and the consequences of this alteration on T cell phenotype. Information gained from these murine studies will provide preliminary data necessary for understanding how RA patients can successfully be treated with A12. PUBLIC HEALTH RELEVANCE: This R01 is a new application in which we use humanized mice bearing DRB1*0101 and DRB1*0401 transgenes and the collagen-induced arthritis (CIA) model to develop a collagen-based immunotherapy.

描述（申请人提供）：类风湿性关节炎是一种关节间关节的慢性炎症性疾病。在初步研究中，我们发现 50% 的 RA 患者对 II 型胶原蛋白 (CII) 产生显着的 T 细胞反应。我们的假设，即抗原驱动的自身免疫过程介导关节损伤，是基于我们的发现，即在我们的开放标签试验中，对关节炎患者口服 CII 导致所测量的细胞因子谱发生有利的改变。这些数据支持这样的观点：对软骨中的 CII 等抗原的自身免疫在该疾病的发病机制中发挥着重要作用。我们使用携带 DRB1*0101 和 DRB1*0401 转基因的人源化小鼠和胶原诱导关节炎 (CIA) 模型来开发基于胶原蛋白的免疫疗法。通过使用增殖和细胞因子测定，我们发现 Tg 小鼠中 DR1 和 DR4 向小鼠 T 细胞呈递的免疫显性决定簇的核心是 CII263-270 (FKGEQGPK)。随后，开发了一种合成的类似肽 CII 263-273 (F263N, E266D) (A12)，发现在自身免疫性关节炎建立后施用时，可诱导 CIA 的深度抑制。基于这些数据，我们提出以下假设：A12 类似肽/APC 复合物与 TCR 的相互作用导致独特的信号传导途径，很可能涉及 Syk 而不是 ZAP 70 的磷酸化。由此产生的信号传导事件级联主要诱导 Th2 细胞因子并最终抑制关节炎。我们提出以下目标：1) 鉴定 A12 激活的 T 细胞信号传导途径，并将其与野生型 CII 肽诱导的途径进行比较。 2) 表征 A12 诱导的抑制性 T 细胞，并确定该选定群体如何抑制对 CII 和自身免疫性关节炎的免疫力。 3)确定翻译后修饰在增强A12类似物抑制CII和关节炎免疫力的功效中的作用。这些研究将确定 A12 触发哪些细胞内信号传导途径、这些信号传导途径与未改变的肽配体和完整 CII 触发的细胞内信号传导途径有何不同，以及这种改变对 T 细胞表型的影响。从这些小鼠研究中获得的信息将为了解 RA 患者如何成功接受 A12 治疗提供必要的初步数据。公共健康相关性：R01 是一项新应用，我们使用携带 DRB1*0101 和 DRB1*0401 转基因的人源化小鼠以及胶原诱导关节炎 (CIA) 模型来开发基于胶原蛋白的免疫疗法。