Adaptive immunity of P. aeruginosa in cystic fibrosis

铜绿假单胞菌在囊性纤维化中的适应性免疫

• 批准号: 7140532
• 负责人: Gregory P Priebe
• 金额: $ 19.6万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-05 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7140532
• 关键词: Pseudomonas aeruginosa; attenuated microorganism; chemokine; clinical research; confocal scanning microscopy; cystic fibrosis; flow cytometry; gene deletion mutation; gene expression; genetically modified animals; human subject; humoral immunity; immune response; immunoglobulin G; immunomodulators; immunopathology; laboratory mouse; live vaccine; lung disorder; microorganism immunology; mucosal immunity; opsonin; passive immunization; respiratory infections

DESCRIPTION (provided by applicant): This revised application is in response to a Program Announcement for Exploratory and Developmental Research Grants for Investigations in Rare Diseases, which is funded through the R21 award mechanism. This project centers on the rare disease cystic fibrosis (CF), with focus on immunity to the bacterial pathogen Pseudomonas aeruginosa. In CF, infection with P. aeruginosa is linked to clinical deterioration. Antibiotics are steadily and alarmingly losing effectiveness due to resistance. The long-term goals of this project are to elucidate the defects in adaptive immunity toward P. aeruginosa that allow colonization to evolve into chronic lung disease in CF patients and to devise new therapies to correct these defects and vaccines to prevent infection. The major hypothesis being evaluated is that the failure to clear P. aeruginosa in CF is partly due to defective epithelial secretion of the chemokine RANTES, which results in ineffective opsonic antibody responses. We plan to investigate this hypothesis by analyzing the systemic and mucosal humoral immunity engendered by live-attenuated vaccine strains of P. aeruginosa delivered intranasally to transgenic CF mice. We have found that although this vaccination strategy can delay oropharyngeal P. aeruginosa colonization in CF mice, overall serum opsonic antibody levels are lower than those induced in wild type mice. In Aim 1, the mechanisms behind defective opsonic antibody production following nasal immunization of transgenic CF mice will be assessed with focus on RANTES expression using cytokine/chemokine protein arrays. Localization of labeled vaccine and expression of RANTES will also be evaluated using flow cytometry and confocal microscopic. In Aim 2, the immune effectors responsible for delayed oropharyngeal colonization of transgenic CF mice following nasal immunization will be characterized using passive immunotherapy. We will also investigate whether the protective efficacy of this immunization approach can be augmented by the use of RANTES as a mucosal adjuvant. Knowledge gained from these studies will elucidate new strategies for the prevention and treatment of P. aeruginosa infections in the setting of CF.

描述（由申请人提供）：此修订后的申请是响应于探索性和发展研究赠款的计划公告，用于稀有疾病的调查，该计划是通过R21奖励机制资助的。该项目以罕见疾病囊性纤维化（CF）为中心，重点是对细菌病原体铜绿假单胞菌的免疫力。在CF中，铜绿假单胞菌感染与临床恶化有关。抗生素由于抵抗力而稳定而令人震惊的失去有效性。该项目的长期目标是阐明对铜绿假单胞菌的适应性免疫的缺陷，该缺陷使定植能够演变成CF患者的慢性肺部疾病，并设计新疗法以纠正这些缺陷和疫苗以防止感染。评估的主要假设是，在CF中未能清除铜绿假单胞菌的部分原因是趋化因子的上皮分泌缺陷，这导致无效的Opsonic抗体反应无效。我们计划通过分析由固定型铜绿假单胞菌的活疫苗菌株产生的全身性和粘膜体液免疫来研究这一假设。我们发现，尽管这种疫苗接种策略可以延迟CF小鼠的咽孢子菌定殖，但总体血清opsonic抗体水平低于野生型小鼠诱导的抗体水平。在AIM 1中，将使用细胞因子/趋化因子蛋白阵列来评估转基因CF小鼠的鼻腔免疫后的Opsonic抗体产生背后的机制。标记的疫苗的定位和RANTES的表达也将使用流式细胞仪和共聚焦显微镜进行评估。在AIM 2中，使用被动免疫疗法来表征转基因CF小鼠的延迟口咽CF小鼠的免疫效应子。我们还将研究这种免疫方法的保护效果是否可以通过使用Rantes作为粘膜佐剂来增强。从这些研究中获得的知识将阐明在CF的情况下预防和治疗铜绿假单胞菌感染的新策略。