The role of exosomes in Pseudomonas Aeruginosa Corneal Infection

项目总结 铜绿假单胞菌(PA)是细菌性角膜炎的主要致病菌。在戴隐形眼镜期间， 主机防御机制遭到破坏。这使得PA可以突破紧密的角膜屏障，感染 否则就是健康的眼睛。最近，外切体被认为是炎症和炎症的主要参与者。 感染。此外，外切酶体正日益被认为是潜在的治疗剂。 我们的初步数据显示，在感染期间，角膜上皮细胞有大量的外切体释放。 由PA提供。这些外切体含有一种独特的蛋白酶、转录调节因子和涉及的蛋白质的混合物。 在免疫调节方面。我们进一步提供的数据表明，这些外切体都能促进中性粒细胞 激活并传递对角膜上皮细胞的保护，防止进一步侵袭。此外，我们还发现 从自体体液中分离出的外切体具有抗菌和免疫调节特性。 基于我们的发现，我们提出了一个初步的假设，即外切体从PA感染的角膜中释放出来 上皮细胞促进先天免疫细胞清除PA，并启动未感染的角膜上皮细胞 预防PA感染。我们进一步提出了第二个假设，即外切体起源于 富含外体的体液含有有效的抗菌和抗炎介质，可以利用 以促进PA在角膜中的清除和疾病的解决。 我们将检验这些假设如下：目标1.建立从PA感染中分离的外切体是如何 角膜上皮细胞和人体体液在体外影响天然免疫细胞。目标2.确定 从感染PA的角膜上皮细胞和人体体液中分离出的外切体如何影响 角膜上皮细胞对PA的体外反应。目的3.确定从中国分离的外切体 自体体液具有保护、抗菌和免疫调节作用 活体兔隐形眼镜模型。 为了完成这些研究，我们组建了一个高度协作的多学科团队，并拥有 以国家最先进的资源在眼科和德克萨斯大学西南的核心设施。这些 研究具有重要意义和创新性，因为它们是第一个在所有 组织或细胞系统。体液外切体的潜在治疗用途代表了一种 治疗角膜感染的重大范式转变，具有广泛的治疗意义。

PROJECT SUMMARY Pseudomonas aeruginosa (PA) is the leading causative agent in microbial keratitis. During contact lens wear, host defense mechanisms are compromised. This allows PA to breach the tight corneal barrier and infect the otherwise healthy eye. Recently, exosomes have been implicated as major players in inflammation and infection. In addition, exosomes are becoming increasingly recognized as potential therapeutic agents. Our preliminary data shows that there is massive exosome release from corneal epithelial cells during infection by PA. These exosomes contain a unique mixture of proteases, transcriptional regulators and proteins involved in immune regulation. We further provide data that indicates that these exosomes both promote neutrophil activation and convey protection to corneal epithelial cells against further invasion. Moreover, we have found that exosomes isolated from autologous body fluids have antimicrobial and immunomodulatory properties. Based on our findings, we propose the primary hypothesis that exosomes released from PA-infected corneal epithelial cells promote PA clearance by innate immune cells and prime non-infected corneal epithelial cells to defend against PA infection. We further propose the secondary hypothesis that exosomes derived from exosome rich body fluids contain potent antimicrobial and anti-inflammatory mediators that can be harnessed to promote PA clearance and disease resolution in the cornea. We will test these hypotheses as follows: Aim 1. Establish how exosomes isolated from PA-infected corneal epithelial cells and human body fluids impact innate immune cells in vitro. Aim 2. Determine how exosomes isolated from PA-infected corneal epithelial cells and human body fluids impact the corneal epithelial response to PA in vitro. Aim 3. Determine whether exosomes isolated from autologous human body fluids exhibit protective antimicrobial and immunomodulatory properties in the rabbit contact lens model in vivo. To accomplish these studies, we have compiled a highly collaborative multidisciplinary team and have access to state of the art resources in the Department of Ophthalmology and UT Southwestern core facilities. These studies are significant and innovative because they are the first of their kind for PA infection in any tissue or cell system. The potential therapeutic use of exosomes from human body fluids represents a major paradigm shift for treating corneal infections and has broad therapeutic implications.