Gene Expression and Pathogenicity of P.aeruginosa

铜绿假单胞菌的基因表达和致病性

• 批准号: 6833956
• 负责人: Jeanine P Wiener-Kronish
• 金额: $ 34.64万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-12-11 至 2007-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6833956
• 关键词: Pseudomonas aeruginosa; bacteria infection mechanism; bacterial genetics; bacterial proteins; clinical research; gel electrophoresis; gene expression; host organism interaction; human subject; laboratory mouse; lung injury; microarray technology; southern blotting; toxin; virulence

DESCRIPTION (provided by applicant): We are proposing a translational investigation, including clinical as well as basic scientists, to test the hypothesis that specific Pseudomonas virulence products determine whether lung infection occurs. The results from the proposed experiments will be used to develop diagnostic tools that can identify virulent Pseudomonas in patients. The results will also identify critical bacterial gene products involved in lung injury. The present application carefully dissects associations between bacterial virulence and lung injury so that future mechanistic investigations in patients can be proposed. Without the data from the proposed investigations, we do not have the information to proceed. We already have Pseudomonas strains obtained from a cohort of intubated, mechanically ventilated patients from another grant. Our first aim is to perform a genomic analysis of Pseudomonas strains obtained from 2 distinct patient populations; intubated, ventilated patients with signs of lung infections that have disseminated and intubated, ventilated patients who have no sign of disease. Using microarray analysis, we will determine the significantly different genes between the Pseudomonas strains found in these two patient groups and produce consensus primers to these genes. We will then utilize these consensus primers in a prospective investigation of 300 intubated, ventilated patients to confirm the prevalence of the genetic differences in colonized vs infected intubated, ventilated patients. The second aim is to compare the in vivo virulence gene expression of Pseudomonas strains obtained from colonized patients to the expression profile of the strains obtained from patients who have Pseudomonas lung infections Analysis of bacterial gene expression in vivo will be done using real-time PCR. The results of all these experiments will allow us to prove (or disprove) that there are different genes or different patterns of toxin gene expression between the Pseudomonas strains infecting patients and those colonizing patients. In the final aim, we will evaluate the interaction of type III toxin genes in vivo By utilizing an isogenic bacterial strain (PA103) that has each of the toxin genes added alone or in combination, we will be able to define the effects of each of the type III toxins as well as other genes we find of significance in the first two aims.

描述（由申请人提供）： 我们提出了一项翻译研究，包括临床和基础科学家，以检验特定假单胞菌毒力产物确定肺部感染是否发生的假设。拟议的实验的结果将用于开发可以鉴定患者有毒假单胞菌的诊断工具。结果还将确定与肺损伤有关的关键细菌基因产物。本应用仔细剖析了细菌毒力与肺损伤之间的关联，因此可以提出患者的未来机械研究。没有拟议调查的数据，我们没有继续进行的信息。我们已经有了从另一份拨款的插管，机械通风的患者中获得的假单胞菌菌株。我们的第一个目的是对从2个不同患者种群获得的假单胞菌菌株进行基因组分析。插管，通风的患者患有肺部感染迹象，这些患者已经传播和插管没有疾病迹象的通风患者。使用微阵列分析，我们将确定在这两个患者组中发现的假单胞菌菌株之间的显着不同基因，并对这些基因产生共识引物。然后，我们将在对300名插管，通风患者的前瞻性研究中利用这些共识引物，以确认定植与感染的插管，通风患者的遗传差异的流行。第二个目的是比较从定殖患者获得的假单胞菌菌株的体内毒力基因表达与从患有假单胞菌肺肺感染分析的细菌基因表达在体内的患者获得的菌株的表达谱，使用实时PCR进行。所有这些实验的结果将使我们能够证明（或反驳）在感染患者的假单胞菌菌株与患者定植的假单胞菌菌株之间存在不同的基因或不同的毒素基因表达。在最终目的中，我们将通过利用同基因细菌菌株（PA103）来评估体内III型毒素基因的相互作用，该菌株具有单独添加或组合的每个毒素基因，我们将能够定义第一个III型毒素以及其他每个基因的效果，我们在第一个两个目标中都具有重要的意义。