Diagnostic Tests and Immunotherapy of Sarcoidosis Using Mycobacterial Proteins

描述（由申请人提供）：结节病是一种多系统肉芽肿性疾病，90%以上的患者累及肺部，可导致终末期肺纤维化和死亡。结节病的发病率为-10-40 / 10万人，主要发生在非裔美国人和美国妇女中，结节病代表了一个重大的健康问题和健康差异问题。结节病的病理特征是非干酪化肉芽肿性炎症。没有结节病的诊断测试，除了对受影响的组织进行活检，这需要相当的费用和风险。结节病被广泛认为诊断不足，部分原因是缺乏诊断工具。目前还没有安全有效的治疗结节病的方法。本应用程序的目的首先是开发一种安全、标准化的结节病诊断皮肤试验，其次是建立一种治疗结节病的免疫治疗方法。我们的具体方法利用了我们最近的发现，即结核杆菌过氧化氢酶（mKatG）是结节病的原型致病组织抗原。这一发现是基于一种新的蛋白质组学方法，其中mKatG是根据Kveim反应的生物物理特性分离出来的，Kveim反应是一种已建立的结节病诊断皮肤试验，由于使用同种异体结节病组织，因此出于安全考虑而停止。假设是1。2. mKatG诱导结节病患者皮肤局部肉芽肿反应，具有疾病相关肉芽肿和Kveim反应的特征，可作为诊断工具；在结节病临床前模型中，口服mKatG疫苗可抑制实验性mKatG驱动的肉芽肿性肺炎症。在Aim 1研究中，我们将根据良好生产规范生产重组mKatG，适合用作皮内皮肤试验试剂，并测试结节病患者和对照受试者皮内给药的安全性和效果。在Aim 2研究中，我们将验证口服mKatG疫苗通过上调mKatG特异性调节性T细胞来抑制实验性肉芽肿性肺炎症的假设，从而建立使用mKatG等原型致病抗原口服疫苗治疗结节病的免疫学框架。相关性（见说明书）：我们提出的研究可能会导致结节病安全、简单的诊断性皮肤试验的发展，从而降低风险、医疗费用和误诊。此外，我们的研究将测试基于疾病相关微生物蛋白的口服疫苗是否能够抑制结节病中所见的特征性炎症，从而为结节病提供一种新颖且可能更安全的治疗策略。

DESCRIPTION (provided by applicant): Sarcoidosis is a multisystem granulomatous disorder that involves the lungs in over 90% of affected individuals and may cause end-stage pulmonary fibrosis and death. With an incidence of -10-40 per 100,000 people, predominant in African Americans and women in the U.S., sarcoidosis represents a significant health problem and health disparity concern. The pathologic hallmark of sarcoidosis is non-caseating granulomatous inflammation. There are no diagnostic tests for sarcoidosis except for biopsy of affected tissues that carry considerable expense and risk. Sarcoidosis is widely believed to be underdiagnosed, in part due to a lack of diagnostic tools. There are no safe, effective treatments for sarcoidosis. The goals of this application are first to develop a safe, standardized diagnostic skin test for sarcoidosis, and second, to establish an immunotherapeutic approach for treatment of sarcoidosis. Our specific approach capitalizes on our recent discovery that IVIycobacterium tuberculosis catalase-peroxidase (mKatG) is a prototypic pathogenic tissue antigen in sarcoidosis. This discovery was based on a novel proteomic approach in which mKatG was isolated based on the biophysical properties of the Kveim reaction, an established diagnostic skin test for sarcoidosis which was discontinued for safety concerns since it uses allogeneic sarcoidosis tissues. Our hypotheses are 1. mKatG induces a local granulomatous response in the skin of sarcoidosis patients characteristic of disease related granulomas and Kveim reactions that can serve as a diagnostic tool, and 2. oral vaccination with mKatG suppresses experimental mKatG driven granulomatous lung inflammation in a preclinical model of sarcoidosis. In Aim 1 studies, we will manufacture recombinant mKatG under Good Manufacturing Practices suitable for use as an intradermal skin test reagent and test the safety and effect of intradermal administration in sarcoidosis patients and control subjects. In Aim 2 studies, we will test the hypothesis that oral vaccination with mKatG suppresses experimental granulomatous lung inflammation by upregulating mKatG specific regulatory T cells, thereby establishing the immunologic framework for treating sarcoidosis with oral vaccines using prototypic pathogenic antigens such as mKatG. RELEVANCE (See instructions): Our proposed studies may lead to the development of a safe, simple diagnostic skin test for sarcoidosis that could reduce risk, health care expenses and misdiagnosis. Furthermore, our studies would test whether an oral vaccine based on disease-related microbial proteins is able to suppress the characteristic inflammation seen in sarcoidosis, offering a novel, and possibly, far safer treatment strategy for sarcoidosis.