Pathogenesis of microbial anterior eye diseases

微生物性眼前部疾病的发病机制

基本信息

批准号：
8930158
负责人：
Gerald B Pier
金额：
$ 41.67万
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 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8930158
关键词：
Affect Anterior Antibodies Applications Grants Aspergillus niger Blindness Candida Candida albicans Cell surface Cells Communities Comparative Genomic Analysis Conjunctivitis Contact Lenses Cornea Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator DNA Sequence Epithelial Cells Evaluation Eye Eye Infections Eye diseases Funding Fusarium Gene Deletion Genes Genome Germ-Free Goals Growth Haemophilus influenzae Health High-Throughput Nucleotide Sequencing Human Immune response Immune system Immunity Immunization Immunotherapy In Vitro Infection Infectious Agent Inflammatory Inflammatory Response Integration Host Factors Investigation Keratitis Lead Ligands Lipopolysaccharides Mediating Medical Membrane Microdomains Methodology Microbe Modeling Molecular Mus Nature Neisseria Neisseria gonorrhoeae Niger Oligosaccharides Organism Panton-Valentine leukocidin Pathogenesis Pathology Polysaccharides Prevention Pseudomonas aeruginosa Research Screening Result Sequence Analysis Staphylococcus aureus Streptococcus pneumoniae Streptococcus pyogenes Surface Technology Testing Therapeutic Tissues Transgenic Organisms Treatment Efficacy Vaccines Validation Virulence base corneal epithelium fitness genome-wide in vivo injured insight microbial mutant pathogen poly-N-acetyl glucosamine pre-clinical prevent prophylactic subcutaneous therapeutic evaluation vaccine evaluation vaccine trial

项目摘要

DESCRIPTION (provided by applicant): The broad, long term goals of this grant application are to understand the microbial factors and the molecular and cellular host responses to pathogens that are significant causes of infections in the anterior eye. These insights should lead to prophylactic and therapeutic strategies that can be clinically assessed for preventing or treating these serious infections. Specifically, this proposal will focus on infections caused by Pseudomonas aeruginosa and the plethora of pathogens causing keratitis and conjunctivitis that express the broadly conserved surface polysaccharide poly-N-acetyl glucosamine (PNAG). PNAG is not expressed by P. aeruginosa but is expressed by most major gram-positive and gram-negative bacterial organisms associated with eye infections including Staphylococcus aureus, S. epidermidis, Streptococcus pneumoniae, S. pyogenes, Hemophilus influenzae and Neisseria gonorrhoeae as well as by serious fungal causes of keratitis including Fusarium spp., Aspergillus niger and Candida albicans. For P. aeruginosa, we have found that this microbe enters corneal epithelial cells both in vitro and during experimental infection of the scratched-injured mouse eye via lipid rafts containing the cystic fibrosis transmembrane conductance regulator (CFTR), and that a large proportion of the bacterial cells are intracellular. The bacteril ligand for CFTR is the outer core oligosaccharide of the cell surface lipopolysaccharide. In the currently-funded proposal, we further defined some of the molecular and cellular factors activated by the P. aeruginosa-CFTR interaction in the cornea leading to destructive inflammatory responses and also analyzed in depth the ability of antibody to P. aeruginosa to prevent pathology and infection in keratitis. We initiated a rigorous study of virulence and immunity to S. aureus corneal infection including the subset that express the Panton-Valentine Leukocidin (PVL) and established the potential for immunotherapy against the PNAG surface polysaccharide to prevent or treat of S. aureus keratitis. In the ensuing years we will extend our long term goals by further analyzing the ability of antibody-mediated immunity to PNAG-expressing organisms to prevent anterior eye infections using models of keratitis and conjunctivitis, one occurring in an avascular tissue and the other in a vascularized ocular tissue. These contrasting settings will allow us to investigate, compare and contrast the host cellular co-factors needed for effective immunity which can be conducted using transgenic and germ-free mice lacking matured immune systems. Additionally, to continue our investigations into the pathogenesis of P. aeruginosa corneal infection we will use high- throughput sequencing analysis of a saturated transposon (Tn) bank of P. aeruginosa that we have recently established as a major approach to the study of the host-pathogen interaction to investigate bacterial and host factors involved in the pathogenesis of keratitis. Accomplishing these goals will provide the pre-clinical justification for testing of immunotherapies to PNAG in human ocular infections and extend the insight into the pathogenesis of P. aeruginosa keratitis to a heretofore unachieved level of precision and detail.

描述（由申请人提供）：本拨款申请的广泛、长期目标是了解微生物因素以及分子和细胞宿主对病原体的反应，这些病原体是前眼感染的重要原因。这些见解应该会导致预防和治疗策略的产生，这些策略可以通过临床评估来预防或治疗这些严重的感染。具体来说，该提案将重点关注铜绿假单胞菌引起的感染以及引起角膜炎和结膜炎的大量病原体，这些病原体表达广泛保守的表面多糖聚-N-乙酰氨基葡萄糖（PNAG）。 PNAG 不由铜绿假单胞菌表达，但由大多数与眼部感染相关的主要革兰氏阳性和革兰氏阴性细菌表达，包括金黄色葡萄球菌、表皮葡萄球菌、肺炎链球菌、化脓性链球菌、流感嗜血杆菌和淋病奈瑟菌以及由引起角膜炎的严重真菌引起，包括镰刀菌属、黑曲霉和念珠菌白色念珠菌。对于铜绿假单胞菌，我们发现这种微生物在体外和在划伤小鼠眼睛的实验感染过程中通过含有囊性纤维化跨膜电导调节剂（CFTR）的脂筏进入角膜上皮细胞，并且大部分细菌细胞是细胞内的。 CFTR 的细菌配体是细胞表面脂多糖的外核寡糖。在目前资助的提案中，我们进一步定义了角膜中铜绿假单胞菌-CFTR相互作用激活的一些分子和细胞因子，导致破坏性炎症反应，并深入分析了铜绿假单胞菌抗体预防病理的能力和角膜炎感染。我们发起了一项针对金黄色葡萄球菌角膜感染（包括表达 Panton-Valentine 白细胞杀素 (PVL) 的子集）的毒力和免疫力的严格研究，并确定了针对 PNAG 表面多糖的免疫疗法预防或治疗金黄色葡萄球菌角膜炎的潜力。在接下来的几年中，我们将通过进一步分析抗体介导的免疫对表达 PNAG 的生物体的能力来扩展我们的长期目标，以使用角膜炎和结膜炎模型（一种发生在无血管组织中，另一种发生在血管化组织中）来预防前眼部感染。眼组织。这些对比设置将使我们能够研究、比较和对比有效免疫所需的宿主细胞辅助因子，这可以使用缺乏成熟免疫系统的转基因和无菌小鼠进行。此外，为了继续研究铜绿假单胞菌角膜感染的发病机制，我们将使用我们最近建立的铜绿假单胞菌饱和转座子（Tn）库的高通量测序分析作为宿主研究的主要方法-病原体相互作用，以研究角膜炎发病机制中涉及的细菌和宿主因素。实现这些目标将为人类眼部感染中 PNAG 免疫疗法的测试提供临床前依据，并将对铜绿假单胞菌角膜炎发病机制的深入了解扩展到迄今为止未达到的精确度和细节水平。