Mechanism of Mycoplasma-Induced Mast Cell Il-4 Synthesis

支原体诱导肥大细胞 IL-4 合成的机制

• 批准号: 7063425
• 负责人: THOMAS PRESCOTT ATKINSON
• 金额: $ 30.07万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7063425
• 关键词: Escherichia coli; Mycoplasma pneumoniae; SDS polyacrylamide gel electrophoresis; adhesin; asthma; cell cell interaction; flow cytometry; genetically modified animals; glycoproteins; helper T lymphocyte; inflammation; interleukin 4; laboratory mouse; mast cell; mycoplasmal pneumonia; polymerase chain reaction; site directed mutagenesis; toll like receptor; western blottings

The overall goals of this project are to define innate and adaptive immune mechanisms by which Mycoplasma pneumoniae contributes to the development of chronic airway hyperreactivity and inflammation, focusing on the role of mast cells. Specific Aim 1: To clone and express the P1 adhesin of M. pneumoniae in E. coli and to produce a mutant M. pneumoniae strain deficient in that molecule. The P1 adhesin or select fragments involved in the mast cell IL-4 response will be cloned and expressed in E. coli. Using transposon mutagenesis, a P1-deficient strain of M. pneumoniae will be produced for use in mast cell stimulation experiments and the mouse model of infection. Specific Aim 2: To identify and characterize surface molecules on mast cells that are essential for induction of IL-4 production by P1 glycoprotein, examine modulating effects of other molecules in the pulmonary microenvironment, validate the effects observed using human mast cells, and characterize the effects of knockdown of expression of specific molecules involved in the M. pneumoniae cytokine response in mast cell lines. Target molecules for M. pneumoniae-induced mast cell cytokine production will be identified and characterized and the effects of mutational deletion examined, modulating effects of mediators in the pulmonary microenvironment will be analyzed, and effects of M. pneumoniae in a new human mast cell line will be studied. Specific Aim 3: To determine if respiratory tract infection by M. pneumoniae induces the recruitment to the airways of TH2 CD4 + lymphocytes, and to determine if this occurs using a mechanism that is dependent on mast cell-derived IL-4 or other mast cell products. The ability of M. pneumoniae to cooperate in the mast cell-induced recruitment of TH2 cells to the lungs and airways will be studied using adoptive transfer of antigen specific T cells. Specific Aim 4: To characterize the effects of infection in a mouse model under conditions in which interactions between M. pneumoniae and mast cells have been abrogated or in which mast cell IL-4 production is prevented. P1 deficient strain(s) of M. pneumoniae will be tested in mice for effects on airway hyperreactivity in a chronic infection model. The role of target molecules on the mast cells for M. pneumoniae-induced cytokine production will be studied in mutant strains of mice.

该项目的总体目标是确定肺炎支原体促进慢性气道高反应性和炎症发展的先天性和适应性免疫机制，重点关注肥大细胞的作用。 具体目标1：在大肠杆菌中克隆并表达肺炎支原体的P1粘附素，并产生缺乏该分子的肺炎支原体突变株。 P1 粘附素或参与肥大细胞 IL-4 反应的选择片段将在大肠杆菌中克隆和表达。利用转座子诱变，将产生 P1 缺陷型肺炎支原体菌株，用于肥大细胞刺激实验和小鼠感染模型。 具体目标 2：识别和表征肥大细胞上的表面分子，这些分子对于 P1 糖蛋白诱导产生 IL-4 至关重要，检查肺部微环境中其他分子的调节作用，验证使用人肥大细胞观察到的效果，并表征肥大细胞系中参与肺炎支原体细胞因子反应的特定分子表达敲低的影响。将鉴定和表征肺炎支原体诱导的肥大细胞细胞因子产生的靶分子，并检查突变缺失的影响，分析肺部微环境中介质的调节作用，以及肺炎支原体在新的人类肥大细胞系中的影响将被研究。 具体目标 3：确定肺炎支原体引起的呼吸道感染是否会诱导 TH2 CD4 + 淋巴细胞募集到气道，并使用依赖于肥大细胞衍生的 IL-4 或其他肥大细胞的机制来确定是否会发生这种情况产品。将使用抗原特异性 T 细胞的过继转移来研究肺炎支原体配合肥大细胞诱导的 TH2 细胞向肺部和气道募集的能力。 具体目标 4： 表征在肺炎支原体和肥大细胞之间的相互作用已被消除或肥大细胞 IL-4 产生被阻止的条件下，小鼠模型中感染的影响。将在小鼠中测试 P1 缺陷型肺炎支原体菌株对慢性感染模型中气道高反应性的影响。将研究靶分子对肺炎支原体诱导的细胞因子产生的肥大细胞的作用 在小鼠的突变品系中。