VIP and Disease Resolution of Bacterial-Induced Ocular Infection

项目总结/摘要 背景铜绿假单胞菌（PA）引起的角膜炎是最常见和破坏性的角膜炎之一 最终导致失明这种机会性革兰氏阴性菌 最为人所知的是在长期接触透镜佩戴者中引起细菌性角膜炎。仅在美国， 微生物角膜炎的发病率为每年25，000 - 30，000例，治疗费用估计为15 -30美元 万 目标/假设。这种威胁视力的疾病在很大程度上是炎症的结果。 免疫系统的免疫应答依赖于免疫细胞的调节， 这些细胞和微环境释放的抗炎因子，以及有效恢复 组织内稳态在这方面，鉴于抗生素耐药性的发生率增加，血管活性药物 肠肽（VIP）是一种由28个氨基酸组成的神经肽， 在一些实施方案中，本发明涉及以抗炎方式影响免疫应答的免疫调节剂。目标是 描述VIP诱导的炎症和先天性免疫的促消退机制， 角膜感染性疾病模型。此外，启动VIP治疗的临床前研究 角膜感染性疾病的治疗。 具体目标。该建议旨在：1）确定VIP对表达/激活 炎症的专门促消退介质（SPM）; 2）检查VIP如何影响gd T细胞， IL-17的产生在驱动炎症消退中的作用;以及3）确立VIP治疗作为一种免疫调节剂的功效。 针对多种PA菌株的细菌性角膜炎的临床相关治疗。 研究设计.将如下诱导眼部感染：将每只动物的右眼划破， 将含有1 × 106 CFU PA的5 mL等分试样局部应用于受伤的角膜表面。疾病 将在实验（VIP处理的B6小鼠）和 对照（PBS处理的B6小鼠和PBS处理的BALB/c小鼠）动物使用许多良好建立的 评估免疫细胞活化状态、脂质介质表达和活化的技术 （脂氧素、消退素、保护素）和其他炎症参数。此外，还将开展研究， 为VIP建立临床前相关的治疗模式（例如，给药方式，启动 感染后的治疗，对PA的细胞毒性和侵袭性菌株的功效）。 冲击该项目在治疗上研究了免疫和神经内分泌之间的相互作用 系统在解决眼部感染和随后的视觉保护中起着至关重要的作用。 神经系统和视力。特别是，拟议的研究将奠定坚实的基础， 与人类细菌性角膜炎治疗的临床前相关性。

Project Summary/Abstract Background. Pseudomonas aeruginosa (PA)-induced keratitis is one of the most common and destructive of bacterial diseases, ultimately culminating in blindness. This opportunistic, Gram-negative organism is best known to cause bacterial keratitis in extended contact lens wearers. In the United States alone the incidence of microbial keratitis is 25,000-30,000 cases annually with cost of treatment estimated at $15-30 million. Objective/Hypothesis. This sight-threatening disease is in large part a consequence of the inflammatory response invoked by the host, which depends on the regulation of immune cells, balance between pro- and anti-inflammatory factors released by these cells and the microenvironment, and effective restoration of tissue homeostasis. In this regard and in light of increasing incidence of antibiotic resistance, vasoactive intestinal peptide (VIP), a 28-amino acid neuropeptide, has been implicated as a potent endogenous immunomodulator that affects the immune response in an anti-inflammatory manner. The goal is to delineate the VIP-induced pro-resolving mechanisms of inflammation and innate immunity using a murine model of corneal infectious disease. Furthermore, initiate pre-clinical studies for VIP as a therapeutic treatment for corneal infectious disease. Specific Aims. This proposal intends to: 1) ascertain the effects of VIP regarding expression/activation of specialized pro-resolving mediators (SPMs) of inflammation; 2) examine how VIP influences gd T cells and the production of IL-17 in driving inflammatory resolution; and 3) establish the efficacy of VIP treatment as a clinically relevant therapy for bacterial keratitis against multiple strains of PA. Study Design. Ocular infection will be induced as follows: the right eye of each animal will be scarified, and a 5 mL aliquot containing 1 x 106 CFU PA will be topically applied to the wounded corneal surface. Disease response and mechanisms of resolution will be compared between experimental (VIP-treated B6 mice) and control (PBS-treated B6 mice and PBS-treated BALB/c mice) animals using a number of well-established techniques to assess the activation state of immune cells, expression and activation of lipid mediators (lipoxins, resolvins, protectins), and other parameters of inflammation. In addition, studies will be carried out to establish pre-clinically relevant treatment modalities for VIP (e.g., modes of drug delivery, initiation of treatment post-infection, efficacy against cytotoxic and invasive strains of PA). Impact. The project examines therapeutically how interactions between the immune and neuroendocrine systems play an essential role in the resolution of ocular infection and subsequent preservation of the visual nervous system and visual acuity. In particular, the proposed studies will establish a solid basis of pre-clinical relevance to human treatment of bacterial keratitis.