VIP and Disease Resolution of Bacterial-Induced Ocular Infection

描述（由申请人提供）：背景。铜绿假单胞菌（PA）引起的角膜炎是最常见和最具破坏性的细菌性疾病之一，最终导致失明。众所周知，这种机会性革兰氏阴性微生物会导致长期佩戴隐形透镜的人患细菌性角膜炎。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1500 - 3000万美元。目标/假设。这种威胁视力的疾病在很大程度上是由宿主引起的炎症反应的结果，这取决于免疫细胞的调节、这些细胞和微环境释放的促炎因子和抗炎因子之间的平衡以及组织稳态的有效恢复。在这方面，鉴于抗生素耐药性发生率的增加，血管活性肠肽（VIP），一种28-氨基酸 神经肽作为一种有效的内源性免疫调节剂，以抗炎的方式影响免疫应答。目的是使用角膜感染性疾病的小鼠模型来描绘VIP诱导的炎症和先天免疫的促消退机制。此外，启动VIP作为角膜感染性疾病治疗方法的临床前研究。具体目标。该提案旨在：1）确定VIP对炎症的特异性促消退介质（SPM）的表达/激活的影响; 2）检查VIP如何影响γ δ T细胞和IL-17的产生以驱动炎症消退; 3）确定VIP治疗作为细菌性角膜炎的临床相关疗法对多种PA菌株的疗效。研究设计.眼部感染将 诱导如下：将每只动物的右眼划破，将含有1 × 106CFUPA的5mL等分试样局部应用于受伤的角膜表面。将使用许多成熟的技术比较实验（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）动物之间的疾病应答和消退机制，以评估免疫细胞的活化状态、脂质介质（脂氧素、消退素、保护素）的表达和活化以及炎症的其他参数。此外，将开展研究，以建立VIP的临床前相关治疗模式（例如，药物递送模式、感染后治疗的开始、对PA的细胞毒性和侵袭性菌株的功效）。冲击该项目研究了免疫和神经内分泌系统之间的相互作用如何在解决眼部感染和随后保护视觉神经系统和视力方面发挥重要作用。特别是，拟议的研究将建立一个坚实的基础，临床前的相关性，人类治疗细菌性角膜炎。

DESCRIPTION (provided by applicant): 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 γδ 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.