Novel Reagents in Mouse Models of Autoimmune COPD

自身免疫性慢性阻塞性肺病小鼠模型中的新型试剂

• 批准号: 8166497
• 负责人: Michael Borchers
• 金额: $ 23.55万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-15 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8166497
• 关键词: Animal Model; Antigens; Autoantibodies; Autoimmune Process; B-Lymphocytes; CD4 Positive T Lymphocytes; Chronic Bronchitis; Chronic Obstructive Airway Disease; Clinical; Clonal Expansion; Complex; Data; Development; Disease; Disease Progression; Epithelial Cells; Exposure to; Functional disorder; Goals; Hybridomas; Immunoglobulin G; Immunoprecipitation; Investigation; Lead; Link; Literature; Long-Term Effects; Lung; Mass Spectrum Analysis; Morbidity - disease rate; Mus; Pathogenesis; Pathology; Pathway interactions; Patients; Persons; Population; Public Health; Pulmonary Emphysema; Quality of life; Reagent; Research; Risk Factors; Role; Self Concept; Serum; Smoking; T-Cell Receptor; T-Lymphocyte; Tissues; Two-Dimensional Gel Electrophoresis; acquired immunity; adaptive immunity; air filter; autoreactive T cell; autoreactivity; cigarette smoke-induced; cigarette smoking; frontier; global health; immune function; innate immune function; innovation; mortality; mouse model; novel; response; smoking cessation; therapeutic development

DESCRIPTION (provided by applicant): Long-term exposure to cigarette smoke (CS) is the major risk factor in the development of chronic obstructive pulmonary disease (COPD). COPD, which includes chronic bronchitis and emphysema, is a major global health burden and morbidity and mortality are expected to expand in the next decades. Recent studies demonstrate oligoclonal expansions of T and B cells in emphysema patients, persistent oligoclonal CD4 T cell expansions following smoking cessation in a mouse model of COPD, and the presence of autoantibodies against lung components in COPD. Presently, it is unknown whether autoreactivity against pulmonary tissue contributes to disease pathogenesis. Given the limitations of associative clinical findings, the development of novel animal models to investigate autoimmune components in COPD is necessary to investigate the mechanisms whereby CS exposure leads to the development of autoreactivity. Our goal is to identify potential lung antigens and isolate autoreactive T cell clones in a mouse model of COPD. The CENTRAL HYPOTHESIS is that long-term exposure to CS induces aberrant expansions of T cells that are reactive against lung components which contribute to tissue destruction and pulmonary remodeling. This hypothesis is supported by preliminary evidence demonstrating i) excessive accumulation of IgG in the lungs of CS-exposed mice, ii) serum from CS- exposed mice recognizes unique antigens in naive lung, and iii) transfer of IgG from CS-exposed mice, leads to an emphysema-like pathology. We will achieve the objective of this proposal by pursuing two aims: 1) Identify relevant antigens that specifically react with IgG from CS-exposed mice. Using serum from CS-exposed mice, we will perform immunoprecipitation against lung antigens followed by 2D gel electrophoresis and mass spectroscopy analyses, and 2) Isolate autoreactive T cell clones that develop in response a mouse model of COPD. We will also generate T cell hybridomas and screen for reactivity against putative antigens. The concept of self-reactive T cells developing in response to CS exposure is supported by extensive preliminary data from our lab and a growing body of literature. Isolating these reagents will allow us to examine the expanded T cell populations in the lung and define their role in the development of pulmonary changes pathognomonic of COPD. Successful completion of this project will lead to novel reagents (novel T cell clones, autoreactive T cell receptor sequences, and putative antigens) which are critically needed to investigate the complex pathophysiology of COPD. A more complete understanding of autoreactive T cell effector function in COPD will open new frontiers into specific approaches towards therapeutic development in this devastating disease. PUBLIC HEALTH RELEVANCE: The proposed research is relevant to public health because it represents a fundamental shift in the investigation of immune function in the context of cigarette smoke exposure. The successful completion of this project is expected to lead to the development of novel reagents that are required to substantively advance the study of autoimmune mechanisms in the progression of disease. This contribution would decrease the morbidity and mortality, and enhance the quality of life in persons exposed to cigarette smoke.

描述（由申请人提供）：长期暴露于香烟烟雾（CS）是慢性阻塞性肺部疾病（COPD）发展的主要危险因素。包括慢性支气管炎和肺气肿在内的COPD是全球重大的健康负担，发病率和死亡率将在未来几十年中扩大。最近的研究表明，在肺气肿患者中，T和B细胞的寡克隆膨胀，在COPD小鼠模型中戒烟后持续的寡克隆CD4 T细胞膨胀以及COPD中肺部的自身抗体存在。目前，尚不清楚针对肺组织的自动反应性是否有助于疾病发病机理。鉴于关联临床发现的局限性，需要开发新的动物模型来研究COPD中自身免疫成分的发展，这对于研究CS暴露导致自动反应性发展的机制是必要的。我们的目标是在小鼠COPD模型中鉴定潜在的肺抗原和分离自动反应性T细胞克隆。中心假设是，长期暴露于CS会引起T细胞的异常膨胀，这些细胞反应针对肺部成分，这有助于组织破坏和肺部重塑。这一假设得到了初步证据的支持，证明I）IgG过度积累了暴露于CS的小鼠的肺中，II）来自CS暴露的小鼠的血清识别出幼稚肺中独特的抗原，iii）IgG从CS暴露的小鼠中转移，导致伴有伴有类似病理的病理学。我们将通过追求两个目的来实现该提案的目标：1）确定相关的抗原，这些抗原与CS暴露的小鼠专门反应。使用暴露于CS的小鼠的血清，我们将对肺抗原进行免疫沉淀，然后进行2D凝胶电泳和质谱分析，以及2）分离自动反应性T细胞克隆，这些T细胞克隆会响应小鼠COPD的响应。我们还将生成T细胞杂交瘤和筛选，以实现针对推定抗原的反应性。我们实验室的广泛初步数据和越来越多的文献体系的广泛初步数据支持了自反应性T细胞为CS暴露而发展的概念。分离这些试剂将使我们能够检查肺中扩展的T细胞群体，并定义它们在COPD肺部变化的发展中的作用。该项目的成功完成将导致新的试剂（新型T细胞克隆，自动反应性T细胞受体序列和推定的抗原），这些试剂迫切需要研究COPD的复杂病理生理学。对COPD中的自动反应性T细胞效应功能的更全面了解将为这种毁灭性疾病的治疗性发育提供新的边界。 公共卫生相关性：拟议的研究与公共卫生有关，因为它代表了在烟烟暴露的背景下对免疫功能的调查的根本转变。预计该项目的成功完成将导致新的试剂的发展，这些试剂实质性地研究了疾病进展中自身免疫机制的研究。这种贡献将降低发病率和死亡率，并提高暴露于香烟烟雾的人的生活质量。