Modulation of cytokine networks in mycoplasma pneumonia

描述（由申请人提供）：本项目的广泛、长期目标是确定对支原体呼吸道疾病发病机制和保护有影响的免疫和炎症反应。在亲本研究中，假设T辅助细胞在确定小鼠支原体呼吸道疾病中有益和有害免疫反应之间的平衡方面起着核心作用。此外，这种平衡是由T细胞亚群介导的，其活性由调节性细胞因子（如IL-4和ifn - γ）调节。事实上，Th细胞具有促炎作用，而CD8+ T细胞出人意料地抑制了支原体感染引起的炎症性疾病。此外，研究结果表明，ifn - γ在抗支原体的先天免疫中起着一种新颖但重要的调节作用。这些结果表明，该疾病涉及的免疫机制是复杂的，需要更广泛地了解疾病发病过程中产生的调节细胞因子以及T细胞和细胞因子对其的调节。因此，目前的研究旨在建立一种更广泛、更探索性的方法来分析细胞因子的产生，是否有助于我们理解它们的调节及其在支原体呼吸系统疾病中的潜在作用。

DESCRIPTION (provided by applicant): The broad, long-term objective of the parent grant (5 R01 AI42075-04) and this project is to determine the immune and inflammatory responses that have an impact on the pathogenesis of and protection from mycoplasma respiratory disease. In the parent grant, the hypothesis is that T helper cells play a central role in determining the balance between helpful and harmful immunologic responses in murine mycoplasma respiratory disease. Furthermore, this balance is mediated by T cell subsets whose activity is modulated by regulatory cytokines, such as IL-4 and IFN-gamma. In fact, Th cells are proinflammatory while CD8+ T cells were found to unexpectedly dampen the inflammatory disease due to mycoplasma infection. In addition, the results demonstrate that IFN-gamma plays a novel but an important regulatory role in innate immunity against mycoplasma. These results indicate that the immune mechanisms involved in this disease are complex and require a broader understanding of the regulatory cytokines produced during disease pathogenesis and their regulation by T cells and cytokines. Thus, the current studies are proposed to establish whether a broader, more exploratory approach to analyze production of cytokines can facilitate our understanding their modulation and their potential role in mycoplasma respiratory disease. The Specific Aims address the following questions: 1) What cytokines are produced in lungs of normal or IFN-gamma deficient mice at different stages of mycoplasma disease?; and 2) What is the impact of CD4+ and CD8+ T cells on pulmonary cytokine production? The methods to be used in this exploratory approach are to first monitor cytokine mRNA production using array technology, and those cytokine mRNAs, shown modulated using arrays, will be further analyzed for their levels of expression and for experimental variations using real-time RT- PCR analysis. Furthermore, cytokine protein arrays will be developed to assess their production in a microarray format; once established tissue homogenates and cell culture supernatants to do this, we will develop assays using pre-existing antibodies to capture and detect cytokine protein in an array format. Thus, by analysis of a broad number of cytokines, clusters of cytokines with similar and/or contrasting regulatory properties should be identified that are associated with disease progression for further functional analysis.