Functional impact of dendritic cell phenotype in a mouse model of myositis

树突状细胞表型对肌炎小鼠模型的功能影响

基本信息

批准号：
7659076
负责人：
DANA P ASCHERMAN
金额：
$ 20.45万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-04-03 至 2011-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7659076
关键词：
Address Adjuvant Adoptive Transfer Antibody Formation Antigen Targeting Antigens Autoantibodies Autoimmune Diseases Autoimmune Responses Autoimmunity Blocking Antibodies Bypass Bystander Suppression Cardiomyopathies Cell Culture Techniques Cell physiology Cells Characteristics Chemicals Clinical Trials Complement Congenic Mice Congenic Strain Connective Tissue Diseases Coupled Dendritic Cells Development Disease Disease model Encephalomyelitis Experimental Autoimmune Encephalomyelitis Future Histidine-tRNA Ligase Human Idiopathic Inflammatory Myopathies Immune System Diseases Immune Targeting Immune response Immunization Immunologics Immunosuppression In Vitro Inflammation Inflammatory Insulin-Dependent Diabetes Mellitus Jo-1 antibody Label Ligase Link Lung Lung Inflammation Lymphocyte Maintenance Measurement Mediating Methods Modeling Mus Muscle Myosin ATPase Myositis Organ Pathogenesis Phenotype Physiologic pulse Population Preparation Primary biliary cirrhosis Process Production Publishing Pulmonary Fibrosis Pyruvate Dehydrogenase Complex Relative (related person)Reliance Role Spleen Staining method Stains Stimulus Structure of parenchyma of lung Syndrome System T-Cell Proliferation T-Lymphocyte Therapeutic Tissues Treatment Protocols Variant Work anergy antigen challenge base clinically significant comparative conditioning cytokine human disease immunoregulation in vivo insight lymph nodes mouse model prevent public health relevance research study response therapeutic target

项目摘要

DESCRIPTION (provided by applicant): In the idiopathic inflammatory myopathies, disease pathogenesis remains largely undefined. However, we have recently published work demonstrating that immunization of different NOD and C57BL/6 congenic strains with murine histidyl-tRNA synthetase (Jo-1) generates muscle and lung inflammation resembling features of the anti-synthetase syndrome. The experimental approach outlined in this application extends these earlier studies through functional manipulation of dendritic cell (DC) phenotype to define characteristics of the Jo-1-specific immune response contributing to disease expression. Specific Aim 1 employs different in vitro cytokine preparations and maturation stimuli to generate Jo-1-loaded DCs capable of skewing toward Th1, Th2, or Th17 immune responses targeting this antigen. Subsequent immunization of NOD and C57BL/6 congenic mice with Jo- 1-pulsed DCs cultivated under defined conditions will highlight the impact of antigen-specific T cell polarization on tissue inflammation in comparison to Jo-1/CFA immunization. Adoptive transfer of organ-infiltrating lymphocytes from DC-immunized to naive mice will firmly establish the relationship between Jo-1-specific autoimmune responses and the anti-synthetase syndrome. Parallel measurement of CFSE-labeled T cell proliferative responses and intracellular cytokine staining of antigen-challenged lymphocytes derived from spleen and organ-draining lymph nodes of DC-immunized mice will further link the observed tissue phenotype to Th17 and/or Th1-polarized, Jo-1-specific T cell responses. In contrast, use of alternative cytokine conditioning in Specific Aim 2 will generate Jo-1-loaded "tolerogenic" DCs capable of blocking disease induction and, more importantly, re-establishing tolerance in existing disease. Complementing the in vivo assessment of Jo-1-pulsed tolerogenic DC immunization in disease preventioNot applicable.melioration, in vitro transwell mixing and antibody blocking experiments will define the relative contribution of anergy, bystander suppression, and contact-mediated inhibition to antigen-specific regulatory T cell function. The mechanistic insight provided by these studies will help to define immunologic targets and therapeutic approaches applicable to future human clinical trials. PUBLIC HEALTH RELEVANCE: This project employs functional manipulation of antigen-pulsed dendritic cells to define the role of Jo-1-specific T cells in a murine model of inflammatory myopathy. This approach will clarify mechanisms of disease initiation/perpetuation and establish the feasibility of antigen- specific, dendritic cell-mediated tolerance induction. The latter objective is relevant to the treatment of multiple autoimmune diseases for which global immunosuppression is the only therapeutic option.

描述（由申请人提供）：在特发性炎症性肌病中，疾病发病机制在很大程度上仍不清楚。然而，我们最近发表的研究表明，用鼠组氨酰-tRNA 合成酶 (Jo-1) 免疫不同的 NOD 和 C57BL/6 同源菌株会产生类似于抗合成酶综合征特征的肌肉和肺部炎症。本申请中概述的实验方法通过树突状细胞 (DC) 表型的功能操作扩展了这些早期研究，以定义有助于疾病表达的 Jo-1 特异性免疫反应的特征。具体目标 1 采用不同的体外细胞因子制剂和成熟刺激来产生负载 Jo-1 的 DC，能够偏向针对该抗原的 Th1、Th2 或 Th17 免疫反应。随后用在规定条件下培养的Jo-1脉冲DC对NOD和C57BL/6同系小鼠进行免疫，与Jo-1/CFA免疫相比，将突出抗原特异性T细胞极化对组织炎症的影响。将 DC 免疫的器官浸润淋巴细胞过继转移至幼鼠体内，将牢固地建立 Jo-1 特异性自身免疫反应与抗合成酶综合征之间的关系。对来自 DC 免疫小鼠的脾脏和器官引流淋巴结的抗原攻击淋巴细胞进行 CFSE 标记的 T 细胞增殖反应和细胞内细胞因子染色的平行测量，将进一步将观察到的组织表型与 Th17 和/或 Th1 极化联系起来。 -1-特异性T细胞反应。相比之下，在Specific Aim 2中使用替代细胞因子调节将产生负载Jo-1的“致耐受性”DC，其能够阻断疾病诱导，更重要的是，能够在现有疾病中重新建立耐受性。补充 Jo-1 脉冲耐受性 DC 免疫在疾病预防中的体内评估不适用。改善、体外 Transwell 混合和抗体阻断实验将确定无反应性、旁观者抑制和接触介导的抑制对抗原特异性调节的相对贡献T细胞功能。这些研究提供的机制见解将有助于确定适用于未来人体临床试验的免疫靶点和治疗方法。公共健康相关性：该项目采用抗原脉冲树突状细胞的功能操作来定义 Jo-1 特异性 T 细胞在炎症性肌病小鼠模型中的作用。该方法将阐明疾病发生/持续的机制，并建立抗原特异性、树突状细胞介导的耐受诱导的可行性。后一个目标与多种自身免疫性疾病的治疗相关，全身免疫抑制是唯一的治疗选择。