Cellular and Molecular Mechanisms that Contribute to Ocular Surface Disease during Severe Allergic Inflammation

摘要 炎症性眼睑缘病是由慢性眼表炎症引起的 美国和国外的健康问题。这种类型的炎症，也被称为眼缘炎，可以影响 包括眼睑边缘的前部和后部，可能包括眉板腺功能障碍(MGD)。 泪膜的长期缺陷以及对结膜和角膜的继发性损伤通常是由于 对于MGD，因为眉板腺负责泪膜所需的油(或蜂蜜)的产生 动态平衡。这种衰弱的眼缘炎经常发生在患有慢性过敏性眼病的患者身上， 如特应性角膜结膜炎(AKC)。同样，眼睑炎发生在眼部酒渣鼻、慢性移植物对 宿主病、史蒂文斯-约翰逊综合征等慢性炎症性眼表疾病。我们的实验室 此前已经建立并验证了严重过敏性眼病(AED)小鼠模型。这种AED模型 可重复地在人类身上产生与AKC一致的各种临床后遗症，包括角膜受累， 球结膜上皮下纤维性变，以及眼缘炎(前部和后部)。这一模型的实用性 已被证明可以识别导致严重眼睛过敏的未知致病事件，这是一种 没有得到满足的医疗需求的疾病。按照这些思路，这笔赠款的目的是阐明免疫学 在AED模型中导致眼缘炎的具体机制。拟议的实验超越了 经典变态反应性炎症检查，如肥大细胞脱颗粒，辅助性T细胞2反应和 嗜酸性粒细胞增多症。相反，我们的初步数据使我们将努力集中在T助手17的核心作用上 路径。这里的实验还试图阐明上游介体，包括鉴定博纳 真性树突状细胞亚群及其介体，在该病中负责引发T辅助分子17途径 模特。我们的方法涉及大量体内基因技术，以实现条件性/诱发性耗竭。 特定的树突状细胞亚群以部位特异性的方式。这些战略和其他战略将使我们能够 明确在AED模型中导致眼睑缘病的介质及其机制。这是及时和 一项新的提议有望促进我们对眼粘膜树突状细胞在疾病和 睑缘炎患者的淋巴细胞活化状态。此外，从这些实验中收集的信息可能是 与眼表炎症性疾病这一更广泛的背景相关。

ABSTRACT Inflammatory lid margin disease, which arises in chronic ocular surface inflammation, constitutes a major health problem in the U.S. and abroad. This type of inflammation, also referred to as blepharitis, can affect both the anterior and posterior aspects of the lid margin, and may include meibomian gland dysfunction (MGD). Long-term defects of the tear film and secondary impairments to the conjunctiva and cornea often develop due to MGD, as the meibomian gland is responsible for the oil (or meibum) production required for tear film homeostasis. Such debilitating blepharitis occurs regularly in patients with chronic forms of allergic eye disease, such as in atopic keratoconjunctivitis (AKC). Likewise, blepharitis occurs in ocular rosacea, chronic graft versus host disease, Stevens-Johnson Syndrome, and other chronic inflammatory ocular surface diseases. Our lab has previously established and validated a severe allergic eye disease (AED) mouse model. This AED model reproducibly develops various clinical sequelae consistent with AKC in humans, including corneal involvement, subepithelial fibrosis of the bulbar conjunctiva, and blepharitis (anterior and posterior). The utility of this model has proven to permit identification of unknown pathogenic events that mediate severe eye allergy, which is a disease with an unmet medical need. Along these lines, the aim of this grant is to elucidate the immunologic mechanisms that specifically cause blepharitis in the AED model. Experiments proposed go beyond the examination of classical allergic inflammation, such as mast cell degranulation, T helper 2 responses and eosinophilia. Instead, our preliminary data has led us to focus our efforts on the central role of the T helper 17 pathway. Experiments herein also seek to elucidate upstream mediators, including identification of the bona fide dendritic cell subsets, and their mediators, responsible for eliciting the T helper 17 pathway in this disease model. Our approach involves a myriad of in vivo genetic techniques to enable conditional/inducible depletion of specific dendritic cell subsets in a site specific manner. These strategies and others will empower us to pinpoint the mediators and their mechanisms that cause lid margin disease in the AED model. This timely and novel proposal is poised to catalyze our understanding of eye mucosal dendritic cells in disease and lymphocyte activation in blepharitis. Furthermore, information gleaned from these experiments could be relevant in the broader context of ocular surface inflammatory disease.