Fibrotic cytokine phenotypes, interstitial lung disease

（申请人的摘要）慢性间质性炎症的引发和维持 肺部炎症，导致纤维化，往往是由于动态 激发剂，炎症介质，白细胞， 和肺的结构细胞独立于各种病因，许多 纤维化间质性肺病似乎具有许多类似的 病理反应，包括各种白细胞的初始激发 细胞群转移到肺，随后的成纤维细胞活化和增殖， 和细胞外基质的沉积。了解细胞和 分子机制，负责成纤维细胞活化， 间质性月球炎症的发生、维持和消退 是本申请的广泛、长期目标。工作 该建议的假设如下：成纤维细胞的恶化 在慢性间质性纤维化的发展过程中， 肺部炎症取决于特定疾病的表现 表型特征为2型细胞因子。这些成纤维细胞 通过合成随后的促纤维化因子， 介质，其导致肺纤维化的永久化。这 假设将通过关注纤维化机制来解决， 在Th 1型（最小纤维化）与Th 2型的发展过程中 肺中的（纤维化）免疫应答。细胞因子生物学的特定领域 这将是评估包括：1）白细胞介素的贡献- 4/白细胞介素-13轴，对成纤维细胞的激活作用; 2）Thl样的作用 细胞因子改变成纤维细胞活化; 3）蛋白质组学和基因组学 评估从1型或2型病变分离的成纤维细胞;和4） 嗜酸性粒细胞和淋巴细胞的贡献，从动物中分离， 持续的Th 2应答，或从1型 反应，以影响体外成纤维细胞活化。动物模型 慢性间质性肺炎症将用于评估 Th 1类和Th 2类细胞因子在肺成纤维细胞增殖中的作用 活化和肺胶原沉积。一些技术将 在本申请中使用的方法包括：使用新的组织阵列， 微阵列和激光捕获显微镜;定量逆转录聚合酶 链反应（RT-P（′-R）-TaqMan; mRNA稳定性;以及使用 用于抗原定位的化学分析和用于定量的ELISA 细胞因子。

(Applicant's Abstract) The initiation and maintenance of chronic interstitial pulmonary inflammation, leading to fibrosis, is often due to dynamic interactions between an inciting agent, inflammatory mediators, leukocytes, and structural cells of the lung. Independent of the various etiologies, many fibrotic interstitial lung diseases appear to possess a number of similar pathologic responses, including an initial elicitation of various leukocyte populations to the lung, subsequent fibroblast activation and proliferation, and deposition of extracellular matrix. Understanding the cellular and molecular mechanisms which are responsible for fibroblast activation during the initiation, maintenance, and resolution of interstitial luna inflammation are the broad, long-term objectives of this application. The working hypothesis of this proposal is the following: an exacerbation of fibroblast activation and tissue fibrosis during the evolution of chronic, interstitial lung inflammation is dependent upon the expression of a specific disease phenotype characterized by type 2 cytokines. These fibroblasts mechanistically contribute to the evolution of lung pathology by synthesizing subsequent pro-fibrotic mediators, which result in the perpetuation of lung fibrosis. This hypothesis will be addressed by focusing on fibrotic mechanisms which occur during the development of a Th1 (minimally fibrotic) versus a Th2 type (fibrotic) immune response in the lung. Specific areas of cytokine biology that will be assessed include: 1) the contribution of the interleukin- 4/interleukin-13 axis, to fibroblast activation; 2) the role of Thl-like cytokines in altering fibroblast activation; 3) a proteomic and genomic assessment of fibroblasts isolated from type 1 or type 2 lesions; and 4) the contribution of eosinophils and lymphocytes, isolated from animals with ongoing Th2 responses, or lymphocytes recovered from animals with type1 responses, to influence fibroblast activation in vitro. Animal models of chronic interstitial lung inflammation will be utilized to assess the contribution of Th1 like and Th2 like cytokines during pulmonary fibroblast activation and lung collagen deposition. A number of techniques will be employed in this application including: the use of novel tissue arrays, microarrays, and laser capture microscopy; quantitative reverse transcription-polymerase chain reaction (RT-P('-R)-TaqMan; mRNA stability; and the use of immunohistochermistry for antigen localization and ELISAs for the quantitation of cytokines.