Contact Lens Wear, Bacteria, and Corneal Homeostasis

隐形眼镜佩戴、细菌和角膜稳态

• 批准号: 9762535
• 负责人: Suzanne MJ FLEISZIG
• 金额: $ 39.25万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9762535
• 关键词: Aging; Automobile Driving; Bacteria; C57BL/6 Mouse; Cell membrane; Cells; Coin; Contact Lenses; Contralateral; Cornea; Corneal Stroma; Corynebacterium; Custom; Data; Dendritic Cells; Disease; Epithelium; Event; Exposure to; Eye; Foundations; Homeostasis; Human; Hydrogels; Image; Imiquimod; Immunohistochemistry; Infection; Infiltration; Inflammasome; Inflammation; Inflammatory Response; Interleukin-17; Keratitis; Knockout Mice; Licensing; Ligands; Liquid substance; Location; Lung; Maintenance; Manufacturer Name; Mechanical Stress; Microbe; Modeling; Morphology; Mus; Myopia; Nerve; Neutrophil Infiltration; Nociceptors; Osmolar Concentration; Pathology; Play; Predisposition; Prevention; Pseudomonas aeruginosa; Publishing; Reperfusion Injury; Research; Resolution; Risk; Role; Signal Transduction; Silicones; Skin; Stress; Surgical sutures; TNFRSF1A gene; TRPV1 gene; Temperature; Testing; Thick; Time; Tissues; Vision; base; cell motility; cofactor; commensal microbes; conjunctiva; cytokine; human subject; in vivo Model; infection risk; interleukin-23; lens; microbial; microbiome; mouse model; neutrophil; preservation; recruit; response; tissue stress; tool; trafficking

Summary: Worn by >40 million people in the USA alone, contact lenses are associated with a risk of sight-threatening infection. With increasing use for myopia prevention and efforts to develop lenses for technological purposes, a surge in lens usage (extended wear specifically) is predicted. Unfortunately, research related to lens-related complications has been hindered by lack of an in vivo model amenable to the many research tools available only for mice. Following a 25-year effort, we have developed a mouse contact lens wear model. This utilizes silicone hydrogel lenses custom made by a contact lens manufacturer to fit mouse eyes, and does not require lid suturing. The lenses enable corneal infection when contaminated with P. aeruginosa, the most common causative agent of contact lens-related keratitis in people. Foundational to understanding contact lens related infection is knowing how a lens impacts the cornea without bacterial inoculation. Preliminary data show the mouse model replicates multiple events occurring during human lens wear, including colonization of the lens with commensal-type bacteria, and a dendritic cell (DC) response within 24 h. Moving beyond what is feasible using human subjects, high resolution time-lapse imaging of mice with fluorescent cell membranes revealed normally not present motile cells resembling neutrophils in the stroma after 6 days of wear. Immunohistochemistry confirmed infiltration of Ly6G+ cells (neutrophil marker). This DC/Ly6G+ response without pathology fits the definition of parainflammation as “a low-grade inflammatory response at an intermediate state between tissue homeostasis and classic inflammation which can be induced by persistent tissue stress…” The three aims of this project will explore; 1) triggers of parainflammation during contact lens wear, 2) how the signal is transduced within the cornea to drive the Ly6G+ cell response, and 3) its significance during microbe challenge. The hypothesis, based on published and preliminary data, is that parainflammation during contact lens wear in mice can be triggered by microbes colonizing the lens, initiating a sequence of events not otherwise occurring in the normally microbiome-free cornea that function to protect it against commensal-type bacteria, but which primes the cornea to respond overly-aggressively to P. aeruginosa. !

概括： 仅在美国，就佩戴了4000万人，隐形眼镜与危及观光的风险有关 感染。随着用于近视预防和开发用于技术目的镜头的努力的越来越多的用途， 预测镜头使用量（特别是延长的磨损）。不幸的是，与镜头有关的研究 由于缺乏可用的许多可用研究工具的体内模型，因此阻碍了并发症 仅适用于老鼠。经过25年的努力，我们开发了鼠标隐形眼镜磨损模型。这利用了 硅胶水凝胶镜片由隐形眼镜制造商定制以适合鼠标的眼睛，不需要盖子 缝合。当被铜绿假单胞菌污染时，镜片可使角膜感染，最常见 与隐形眼镜相关角膜炎的病因。 了解隐形眼镜相关感染的基础是知道镜头如何影响角膜 细菌接种。初步数据显示，鼠标模型复制了在 人晶状体磨损，包括具有共生型细菌的晶状体和树突状细胞（DC）的定植 24小时内响应。超越了使用人类受试者可行的东西，高分辨率延时 荧光细胞膜的小鼠成像通常不存在类似的母细胞 磨损6天后，基质中的中性粒细胞。免疫组织化学证实了LY6G+细胞的浸润 （中性粒细胞标记）。这种无病理的DC/LY6G+反应符合副炎症的定义为“ 在组织稳态和经典之间的中间状态下的低度炎症反应 持续的组织应激可以引起的炎症……” 该项目的三个目标将探索； 1）隐形眼镜磨损期间的副炎症触发器，2） 信号在角膜内翻译以驱动LY6G+细胞响应，3）其在微生物中的重要性 挑战。基于已发布和初步数据的假设是接触过程中的副炎症 小鼠的镜头磨损可以通过定植镜头的微生物触发，启动一系列事件，不可以其他方式 发生在通常不含微生物组的角膜中，可保护其免受共生型细菌的影响， 但是，哪些角膜对铜绿假单胞菌的反应过度反应。 呢