Pathogenesis of P. aeruginosa corneal infection

铜绿假单胞菌角膜感染的发病机制

基本信息

批准号：
7172231
负责人：
Gerald B Pier
金额：
$ 39.59万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-01-01 至 2008-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7172231
关键词：
Alginates Antibodies Apoptosis Applications Grants Bacteria Cell Line Cell membrane Cell surface Cells Clinical Complement Cornea Corneal Ulcer Cyclodextrins Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Data Development Disease Disruption Epithelial Cells Evaluation Eye Eye Infections Genes Goals Host Defense Human Immune Immune response Immunity Immunologics Immunotherapeutic agent In Vitro Infection Inflammatory Response Interferon Type II Interleukin-1 Interleukin-18 Interleukin-6 Intervention Keratitis Lead Ligands Lipopolysaccharides Mediating Mediator of activation protein Membrane Microdomains Modality Modeling Molecular Molecular Analysis Monoclonal Antibodies Mus Numbers Oligosaccharides Outcome Pathogenesis Pathologic Pathologic Processes Pathology Patients Phagocytes Polysaccharides Predisposition Prevention Proteins Pseudomonas aeruginosa Reagent Receptor Cell Recording of previous events Resistance to infection Role Site Testing Therapeutic Tissues Transgenic Mice Transgenic Organisms Treatment Efficacy Treatment outcome Wild Type Mouse Work acid sphingomyelinase antimicrobial base chemokine corneal epithelium cystic fibrosis patients cytokine human monoclonal antibodies improved in vivo injured mucoid mutant pathogen prevent receptor response therapy development

项目摘要

DESCRIPTION: The long-term goal of this application is to understand the molecular and cellular responses of corneal epithelial cells to Pseudomonas aeruginosa and develop chemotherapeutic and immunotherapeutic interventions for treatment of this serious eye infection. P. aeruginosa enters corneal epithelial cells via the cystic fibrosis transmembrane conductance regulator (CFTR), and these intracellular bacteria are able to persist in the tissue and avoid host defenses, leading to serious corneal pathology. Goals of aim 1 include exploring the role of the CFTR-P. aeruginosa interactions in corneal pathology as manifest by host chemokine and cytokine responses. Using isogenic human corneal epithelial cell lines expressing either mutant or wild-type (WT) CFTR, P. aeruginosa induced and CFTR-dependent release of cytokines implicated in pathogenesis and resistance to infection, notably IL-1, IL-6, IL-18 and IFN-gamma, will be examined. The role of the identified factors in eye infection and pathology will be tested in appropriate WT and transgenic mice using a corneal scratch-injured eye infection model or in mice in which the factor is neutralized with antibody. P. aeruginosa- CFTR interactions depend on formation of lipid rafts, and preliminary data indicate disruption of rafts prevents corneal pathology and promotes bacterial clearance from infected eyes. In aim 2, the role of these rafts in pathogenesis will be further evaluated using the WT and CF corneal cell lines as well as mice unable to form lipid rafts due to disruption of the acid sphingomyelinase gene, and in mice treated with cyclodextrin, which disrupts lipid rafts. Cyclodextrin treatment of P. aeruginosa eye infections has the potential to be highly efficacious and become an important component of therapy. Finally, a fully human monoclonal antibody (MAb) has been developed to P. aeruginosa alginate a conserved, cell surface polysaccharide that is expressed at low levels by corneal isolates. Importantly, alginate expression is detected n the corneas of P. aeruginosa infected mice and the MAb is highly protective against P. aeruginosa infection. In aim 3, the Mab will be evaluated for therapeutic efficacy against 6 different P. aeruginosa strains in the murine corneal infection model with the goal of having a single immunotherapeutic reagent useful against most clinical isolates as an adjunct to current treatment modalities for P. aeruginosa ulcerative keratitis. Overall at the conclusion of these aims, significant advances in the development of therapies for P. aeruginosa keratitis should become apparent based on use of cellular and molecular studies on host-pathogen interactions.

描述：本申请的长期目标是了解角膜上皮细胞对铜绿假单胞菌的分子和细胞反应，并开发化疗和免疫治疗干预措施来治疗这种严重的眼部感染。铜绿假单胞菌通过囊性纤维化跨膜电导调节器（CFTR）进入角膜上皮细胞，这些细胞内细菌能够持续存在于组织中并逃避宿主防御，导致严重的角膜病变。目标 1 的目标包括探索 CFTR-P 的作用。铜绿假单胞菌在角膜病理学中的相互作用通过宿主趋化因子和细胞因子反应来体现。使用表达突变型或野生型 (WT) CFTR 的同基因人角膜上皮细胞系，铜绿假单胞菌诱导和 CFTR 依赖性释放参与发病机制和感染抵抗力的细胞因子，特别是 IL-1、IL-6、IL-18和 IFN-γ，将被检查。所确定的因素在眼部感染和病理学中的作用将在适当的野生型和转基因小鼠中使用角膜划伤损伤的眼部感染模型或在用抗体中和该因素的小鼠中进行测试。铜绿假单胞菌-CFTR相互作用取决于脂筏的形成，初步数据表明脂筏的破坏可以防止角膜病理并促进受感染眼睛中的细菌清除。在目标 2 中，将使用 WT 和 CF 角膜细胞系以及由于酸性鞘磷脂酶基因破坏而无法形成脂筏的小鼠，以及用环糊精治疗的小鼠，进一步评估这些脂筏在发病机制中的作用。脂筏。环糊精治疗铜绿假单胞菌眼部感染有可能非常有效，并成为治疗的重要组成部分。最后，我们开发了一种全人单克隆抗体 (MAb)，用于铜绿假单胞菌藻酸盐，这是一种保守的细胞表面多糖，在角膜分离株中低水平表达。重要的是，在铜绿假单胞菌感染小鼠的角膜中检测到藻酸盐表达，并且 MAb 对铜绿假单胞菌感染具有高度保护作用。在目标 3 中，将评估 Mab 在小鼠角膜感染模型中针对 6 种不同铜绿假单胞菌菌株的治疗效果，目标是拥有一种可用于大多数临床分离株的单一免疫治疗试剂，作为当前铜绿假单胞菌治疗方式的辅助手段溃疡性角膜炎。总体而言，根据这些目标的结论，基于对宿主-病原体相互作用的细胞和分子研究的使用，铜绿假单胞菌角膜炎疗法的开发方面的重大进展应该变得显而易见。