INFLAMMATORY MECHANISMS IN VIRAL OTITIS MEDIA

急性中耳炎（AOM）和病毒性上呼吸道感染（URIs）代表影响人类群体的两种最常见的疾病，并且造成大量患者发病率和医疗保健成本。 流行病学和实验研究表明，URI在AOM的发病机制中起着因果作用。具体而言，病毒可以侵入中耳（ME）空间并引发炎症反应，最终导致ME渗出（液体）形成和随之而来的症状（疼痛，听力损失）。 病毒暴露后导致人ME炎症反应的分子事件尚未表征。 使用鼻和气管支气管组织的病毒接种的研究表明，起始的炎症事件是通过局部细胞因子产生的上皮介导的。 由于ME通常不含白细胞聚集体，因此病毒性中耳炎的起始事件也可能是上皮介导的。 本研究的长期目标是研究ME粘膜对病毒暴露的反应，以更好地了解AOM的起始事件。 这些知识将有助于开发针对这种重要疾病的新疗法。 该项目的目标是：（1）在实验室的正常ME上皮细胞培养系统中进行甲型流感病毒攻击实验，并使用逆转录酶-聚合酶链反应（RT-PCR）、原位杂交测量白细胞介素（IL）-1 α、IL-1 β、IL-6、IL-8、IL-10和肿瘤坏死因子（TNF）-α基因的表达，和酶联免疫吸附试验（ELISA）;（2）评价来自“耳炎易感”个体的ME上皮细胞中相同的细胞因子应答;和（3）评价几种潜在抑制物质在这些模型中的功效。 这些实验的结果将提供一个更好的了解在病毒性中耳炎的起始事件，并希望，将导致这种疾病的新疗法的发展。主要研究者（PI）成功完成该奖项将为未来作为独立研究者的大型研究提供额外的研究培训和职业发展。

Acute otitis media (AOM) and viral upper respiratory tract infections (URIs) represent the two most common diseases affecting the human population, and account for substantial patient morbidity and health care costs. Epidemiological and experimental studies suggest that URIs play a causal role in the pathogenesis of AOM. Specifically, viruses can invade the middle ear (ME) space and invoke an inflammatory response that culminates in ME effusion (fluid) formation and consequent symptoms (pain, hearing loss). The molecular events responsible for the inflammatory response of the human ME following viral exposure have not been characterized. Studies using viral inoculation of nasal and tracheobronchial tissues suggest that the initiating inflammatory events are epithelium-mediated through local cytokine production. Since the ME normally does not contain aggregates of leukocytes, it is likely that the initiating events in viral otitis media are epithelium-mediated as well. The long-term goal of the present study is to investigate the ME mucosal response(s) to viral exposure in an effort to better understand the initiating events in AOM. Such knowledge will help develop new therapies for this significant disease. The goals of this project are to: (1) perform influenza A viral challenge experiments in the laboratory's normal ME epithelial cell culture system and measure expression of the interleukin (IL,)-1alpha, IL-1beta, IL-6, IL-8, IL-10 and tumor necrosis factor (TNF)-alpha genes using reverse transcriptase- polymerase chain reaction (RT-PCR), in situ hybridization, and enzyme-linked immunoabsorbent assays (ELISA); (2) evaluate the same cytokine responses in ME epithelial cells derived from "otitis-prone" individuals; and (3) evaluate the efficacy of several potentially inhibitory substances in these models. The results of these experiments will provide a better understanding of the initiating events in viral otitis media and, hopefully, will lead to the development of novel therapies for this disease. Successful completion of this award by the principal investigator (PI) will provide the added research training and career development for future large-scale studies as an independent investigator.