Novel Reagents in Mouse Models of Autoimmune COPD

描述(由申请人提供)：长期接触香烟烟雾(CS)是发展为慢性阻塞性肺疾病(COPD)的主要危险因素。包括慢性支气管炎和肺气肿在内的慢性阻塞性肺病是全球主要的健康负担，预计未来几十年的发病率和死亡率将会扩大。最近的研究表明，肺气肿患者的T和B细胞出现寡克隆性扩张，COPD小鼠模型中戒烟后持续的CD4T细胞寡克隆性扩张，以及COPD存在针对肺成分的自身抗体。目前，针对肺组织的自身反应性是否与疾病的发病机制有关尚不清楚。鉴于相关临床研究的局限性，开发新的动物模型来研究COPD的自身免疫成分对于研究CS暴露导致自身反应性发展的机制是必要的。我们的目标是在COPD小鼠模型中识别潜在的肺抗原并分离自身反应性T细胞克隆。中心假设是，长期暴露在CS中会导致T细胞的异常扩张，这种T细胞对肺成分产生反应，从而导致组织破坏和肺重塑。这一假说得到以下初步证据的支持：1)CS暴露小鼠肺组织中的免疫球蛋白过度蓄积，2)CS暴露小鼠的血清识别初生肺中独特的抗原，3)CS暴露小鼠的免疫球蛋白转移，导致肺气肿样病理。我们将通过两个目标来实现这一建议的目标：1)从CS暴露的小鼠中寻找与免疫球蛋白特异性反应的相关抗原。使用CS暴露的小鼠的血清，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，并2)分离作为反应的自身反应性T细胞克隆，以响应COPD小鼠模型。我们还将产生T细胞杂交瘤，并筛选对假定抗原的反应性。来自我们实验室的大量初步数据和越来越多的文献支持了自我反应性T细胞对CS暴露的反应发展的概念。分离这些试剂将使我们能够检查肺中扩大的T细胞群，并确定它们在COPD病因性肺变化发展中的作用。该项目的成功完成将带来新的试剂(新的T细胞克隆、自身反应性T细胞受体序列和假定的抗原)，这些试剂对于研究COPD的复杂病理生理是至关重要的。对COPD自身反应性T细胞效应器功能的更全面了解将为这种毁灭性疾病的治疗开发开辟新的前沿。

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.