MOLECULAR PATHOLOGY OF PNEUMOCYSTIS PNEUMONIA

肺孢子虫肺炎的分子病理学

• 批准号: 6313359
• 负责人: CHAO-HUNG LEE
• 金额: $ 25.55万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-30 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6313359
• 关键词: Pneumocystis carinii; Pneumocystis pneumonia; alveolar macrophages; antisense nucleic acid; cell differentiation; fibronectins; fungal proteins; host organism interaction; laboratory rat; phagocytosis; protein sequence; pulmonary surfactants; transcription factor; transfection /expression vector; vitronectin

DESCRIPTION: Alveolar macrophages from Pneumocystis carinii-infected hosts are defective in phagocytosis, and the expression of the transcription factor GATA-2 in these cells is severely down-regulated. Introduction of a GATA-2-specific antisense oligonuceotide into alveolar macrophages from normal uninfected animals also resulted in a decrease in the phagocytic activity of these cells. In this proposed study, experiments will be performed to further investigate the role of GATA-2 in alveolar macrophage phagocytosis. A GATA-2 expression vector will be introduced into alveolar macrophages from P. carinii-infected hosts to investigate whether GATA-2 over-expression could correct the defect. To further understand the involvement of GATA-2 in P. carinii pathogenesis, genes that are regulated by GATA-2 in monocytes will be identified. DNA microarrays will be probed with labeled cDNA from wild type and GATA-2 knockout monocytes. The hybridized microarrays will then be analyzed to determine which genes are regulated by GATA-2. Experiments will also be performed to determine whether genes such as those encoding MMR, MIP-1alpha, MCP-1, IL6 and TNF-alpha that are associated with functions of alveolar macrophages are regulated by GATA-2. The mechanisms by which P. carinii causes down regulation of GATA-2 in alveolar macrophages will be explored. Normal alveolar macrophages will be incubated directly or indirectly with live or dead P. carinii organisms to determine whether a P. carinii protein is responsible for down regulation of GATA-2 transcription leading to a reduction in the phagocytic activity of alveolar macrophages. Proteins such as vitronectin, fibronectin, surfactant proteins A and D, and P. carinii major surface glycoprotein, which are known to interact with alveolar macrophages during P. carinii infection, will also be examined. These proteins will be incubated either alone or in combination with normal macrophages to determine whether they have any effect on GATA-2 down regulation. Different fractions of bronchoalveolar lavage fluids from P. carinii-infected lung will also be tested. Any P. carinii or host protein that is found to have the ability to down regulate GATA-2 expression will be identified by sequencing a portion of the protein.

描述：来自卡氏肺囊虫感染宿主的肺泡巨噬细胞 吞噬作用和转录因子表达缺陷 这些细胞中的 GATA-2 严重下调。介绍一个 GATA-2特异性反义寡核苷酸进入正常肺泡巨噬细胞 未感染的动物也导致吞噬细胞的活性下降 这些细胞。在这项拟议的研究中，将进行实验以进一步 研究 GATA-2 在肺泡巨噬细胞吞噬作用中的作用。 GATA-2 表达载体将被引入 P. 的肺泡巨噬细胞中。 carinii 感染的宿主研究 GATA-2 过度表达是否可以 纠正缺陷。为了进一步了解 GATA-2 在 P. carinii 发病机制中，单核细胞中受 GATA-2 调控的基因将 确定。 DNA 微阵列将用来自野生型和 GATA-2 敲除单核细胞。然后对杂交的微阵列进行分析 确定哪些基因受 GATA-2 调节。实验也将 进行以确定基因，例如编码 MMR、MIP-1α 的基因， 与肺泡功能相关的 MCP-1、IL6 和 TNF-α 巨噬细胞受 GATA-2 调节。卡氏疟原虫引起的机制 将探讨肺泡巨噬细胞中 GATA-2 的下调。普通的 肺泡巨噬细胞将直接或间接与活的或死的巨噬细胞一起孵育 卡氏疟原虫生物体以确定卡氏疟原虫蛋白是否负责 下调 GATA-2 转录，导致 肺泡巨噬细胞的吞噬活性。蛋白质如玻连蛋白、 纤连蛋白、表面活性蛋白 A 和 D、以及 P. carinii 主表面 糖蛋白，已知其在 P 期间与肺泡巨噬细胞相互作用。 卡里尼感染，也将进行检查。这些蛋白质将被孵化 单独或与正常巨噬细胞结合以确定是否 它们对 GATA-2 下调有任何影响。的不同分数 来自卡氏疟原虫感染肺部的支气管肺泡灌洗液也将被 已测试。任何被发现有能力的卡氏疟原虫或宿主蛋白 下调 GATA-2 表达将通过对部分基因进行测序来鉴定 蛋白质。