FIBROTIC CYTOKINE PHENOTYPES IN INTERSTITIAL LUNG

慢性间质性肺动脉高压的发生和维持 炎症通常是由于刺激性炎症和炎症反应之间的动态相互作用。 剂和肺的结构细胞。 在临床上， 可用于治疗这些疾病的方式是有限的， 可能反映了对病理生理学的理解不足， 调节这些疾病的机制。独立于各种 病因，许多慢性间质性肺疾病似乎具有 许多类似的病理反应，包括初始诱发 不同的白细胞群进入肺，随后的成纤维细胞 活化和增殖以及细胞外基质的沉积。 了解细胞和分子机制， 在启动、维持和 间质性肺炎的解决是广泛的，长期的 本申请的目的。 这项提议的工作假设是， 是以下情况：成纤维细胞活化和组织的加剧 慢性纤维化的演变过程中。肺间质炎症 取决于特定疾病表型的表达 其特征在于Th 2型细胞因子占优势。 这一假设 将通过关注发生在手术期间的纤维化机制来解决。 Th 1（最小纤维化）与Th 2型的发展 肺中的（纤维化）免疫应答。细胞因子的特定区域 将被评估的生物学包括：1）-Th 2样的贡献， 细胞因子，特别是白细胞介素-4、白细胞介素-5和白细胞介素-10， 成纤维细胞活化导致肺纤维化; 2） Thl样细胞因子，特别是白细胞介素-12和γ干扰素，在 在肺纤维化期间改变成纤维细胞活化; 3）a 细胞因子对间质细胞凋亡的作用机制分析 通过腺病毒/细胞因子cDNA转染研究的炎症; 4） 从Th 1或Th 2分离的活化成纤维细胞的贡献 肺损伤到进行性肺部炎症;和5）表达和 调节人类患者中的Th 2样细胞因子谱， 与肺间质纤维化的相关性。 慢性胰腺炎动物模型 将利用间质性肺炎症来评估 Th 1类和Th 2类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。一些 在本申请中将采用各种技术，包括使用 北方印迹、逆转录-聚合酶链反应（RT-PCR）、 mRNA稳定性和原位杂交分析，用于研究 基因表达的调节;免疫组织化学用于 用于定量细胞因子的抗原定位和ELISA， 细胞因子活性的内源性介质;基因转染策略 使用含有鼠IL-4，IL-5，IL-10，γ IFN， 或IL-12 cDNA盒和用于评估功能活性的生物测定 具体的调解人。

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.