Novel Reagents in Mouse Models of Autoimmune COPD

描述（由申请方提供）：长期暴露于香烟烟雾（CS）是慢性阻塞性肺疾病（COPD）发展的主要风险因素。COPD包括慢性支气管炎和肺气肿，是一个主要的全球健康负担，预计发病率和死亡率在未来几十年将增加。最近的研究表明肺气肿患者T和B细胞的寡克隆扩增，COPD小鼠模型戒烟后持续的寡克隆CD4 T细胞扩增，以及COPD中存在针对肺成分的自身抗体。目前，对肺组织的自身反应性是否有助于疾病的发病机制尚不清楚。鉴于相关临床研究结果的局限性，有必要开发新的动物模型来研究COPD中的自身免疫成分，以研究CS暴露导致自身反应性发展的机制。我们的目标是在COPD小鼠模型中鉴定潜在的肺抗原并分离自身反应性T细胞克隆。中枢假设是长期暴露于CS诱导T细胞异常扩增，T细胞对肺成分具有反应性，从而导致组织破坏和肺重塑。该假设得到初步证据的支持，初步证据表明i）IgG在CS暴露小鼠的肺中过度积累，ii）来自CS暴露小鼠的血清识别幼稚肺中的独特抗原，和iii）来自CS暴露小鼠的IgG转移，导致肺气肿样病理。我们将通过追求两个目标来实现本提案的目标：1）鉴定与CS暴露小鼠的IgG特异性反应的相关抗原。使用来自CS暴露小鼠的血清，我们将进行针对肺抗原的免疫沉淀，随后进行2D凝胶电泳和质谱分析，以及2）分离响应于COPD小鼠模型而发展的自身反应性T细胞克隆。我们还将产生T细胞杂交瘤并筛选针对推定抗原的反应性。自我反应性T细胞在CS暴露中的发展的概念得到了我们实验室广泛的初步数据和越来越多的文献的支持。分离这些试剂将使我们能够检查肺中扩增的T细胞群，并确定它们在COPD特异性肺变化发展中的作用。该项目的成功完成将导致新的试剂（新的T细胞克隆，自身反应性T细胞受体序列和推定的抗原），这是研究COPD复杂的病理生理学所急需的。更全面地了解COPD中自身反应性T细胞效应器功能将为这种毁灭性疾病的治疗开发开辟新的领域。

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.