Modulation of cytokine networks in mycoplasma pneumonia

描述（由申请人提供）：母基金（5 R 01 AI 42075 -04）和本项目的广泛、长期目标是确定对支原体呼吸道疾病发病机制和预防有影响的免疫和炎症反应。在母基金中，假设T辅助细胞在决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中起着核心作用。此外，这种平衡由T细胞亚群介导，T细胞亚群的活性由调节细胞因子如IL-4和IFN-γ调节。事实上，Th细胞是促炎性的，而CD 8 + T细胞被发现出乎意料地抑制由于支原体感染引起的炎性疾病。此外，结果表明，IFN-γ在抗支原体的先天免疫中起着新的但重要的调节作用。这些结果表明，参与这种疾病的免疫机制是复杂的，需要更广泛的了解疾病发病过程中产生的调节细胞因子及其调节T细胞和细胞因子。因此，目前的研究建议建立一个更广泛的，更探索性的方法来分析生产的细胞因子，可以促进我们了解他们的调制和他们在支原体呼吸道疾病的潜在作用。 具体目的解决以下问题：1）在支原体疾病的不同阶段，正常或IFN-γ缺陷小鼠的肺中产生哪些细胞因子？2）CD 4+和CD 8 + T细胞对肺细胞因子产生的影响是什么？在该探索性方法中使用的方法是首先使用阵列技术监测细胞因子mRNA的产生，并且将使用实时RT-PCR分析进一步分析那些使用阵列显示调节的细胞因子mRNA的表达水平和实验变异。此外，将开发细胞因子蛋白阵列，以评估其在微阵列格式中的生产;一旦建立了组织匀浆和细胞培养上清液，我们将开发使用预先存在的抗体捕获和检测细胞因子蛋白阵列格式的测定。因此，通过分析大量的细胞因子，应鉴定与疾病进展相关的具有相似和/或相反调节特性的细胞因子簇，用于进一步的功能分析。

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.