FIBROTIC CYTOKINE PHENOTYPES IN INTERSTITIAL LUNG

慢性间质性肺损伤的发生与维持 炎症通常是由于煽动之间的动态相互作用 毒剂和肺的结构细胞。临床上，治疗性的 可用于治疗这些疾病的方法是有限的， 可能反映了对病理生理学的了解不足。 调节机制--这些紊乱。独立于各种 病因，许多慢性间质性肺疾病似乎具有 类似的病理反应的数量，包括最初的诱发 不同的白细胞群到肺，随后成纤维细胞 细胞外基质的活化、增殖和沉积。 了解细胞和分子机制 用于成纤维细胞在启动、维持和 肺间质炎症的消退是广泛的、长期的 此应用程序的目标。这一提议的工作假设 以下是：成纤维细胞激活和组织恶化 慢性纤维化的演变过程。间质性肺炎 取决于特定疾病表型的表达 以Th2型细胞因子为主的。这一假设 将通过关注发生在以下情况的纤维化机制来解决 Th1(轻度纤维化)与Th2型的发展 (纤维化)肺内的免疫反应。细胞因子的特定区域 将被评估的生物学包括：L)-类Th2的贡献 细胞因子，特别是白介素4、白介素5和白介素10， 2)成纤维细胞活化导致肺纤维化的作用 Th1样细胞因子，特别是IL-12和γ干扰素，在 在肺纤维化过程中改变成纤维细胞的激活；3)a 细胞因子对间质作用的机制分析 腺病毒/细胞因子c DNA转染性炎症研究；4) 从Th1和Th2中分离的活化成纤维细胞的作用 肺损害到进行性肺部炎症；5)表达和 人外周血中Th2样细胞因子表达的调节 与间质性肺纤维化的相关性。慢性阻塞性肺疾病动物模型 将利用间质性肺炎症来评估 Th1样细胞因子和Th2样细胞因子在肺损伤中的作用 成纤维细胞活化和肺胶原沉积。一批 将在此应用程序中使用技术，包括使用 Northern印迹、逆转录聚合酶链式反应(RT-PCR)、 MRNA的稳定性和原位杂交分析用于研究 基因表达的调节；免疫组织化学的应用 抗原定位和ELISA定量检测细胞因子和 细胞因子活性的内源性介体；基因导入策略 使用腺病毒，该病毒含有小鼠IL-4、IL-5、IL-10、γ-干扰素， 或IL-12 cDNA盒和评价功能活性的生物测定法 特定的调解人。

The initiation and maintenance of chronic interstitial pulmonary inflammation is often due to dynamic interactions between an inciting agents, and structural cells of the lung. Clinically, the therapeutic modalities that are available to treat these diseases are limited, which likely reflects an insufficient understanding of the pathophysiologic mechanisms that mediate-these disorders. Independent of the various etiologies, many chronic 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 lung 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 the predominance of Th2-type cytokines. This hypothesis will be addressed by focusing on fibrotic mechanisms which occur during the development of a Thl (minimally fibrotic) versus a Th2 type (fibrotic) immune response in the lung. Specific areas of cytokine biology that w:Ill be assessed include: l)- the contribution of Th2-like cytokines, especially interleukin-4, interleukin-5, and interleukin-10, to fibroblast activation leading to pulmonary fibrosis; 2) the role of Thl like cytokines, especially interleukin-12 and gamma interferon, in altering fibroblast activation during pulmonary fibrosis; 3) a mechanistic analysis of the contribution of cytokines to interstitial inflammation via adenovirus/cytokine cDNA transfection studies; 4) the contribution of activated fibroblasts isolated from either Th1 or Th2 lung lesions to progressive lung inflammation; and 5) the expression and regulation of Th2-like cytokine profiles in human patients and correlations with interstitial lung fibrosis. 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 Northern blot, reverse transcription-polymerase chain reaction (RT-PCR), mRNA stability, and in situ hybridization analyses for studying the regulation of gene expression; the use of immunohistochemistry for antigen localization and ELISAs for the quantitation of cytokines and endogenous mediators of cytokine activity; gene transfection strategies using adenovirus containing either a murine IL-4, IL-5, IL-10, gammaIFN, or IL-12 cDNA cassette and bioassays for assessing functional activities of specific mediators.