Dual oxidase and lactoperoxidase in influenza infection

项目总结说明 流感病毒感染每年影响全球数百万人，并导致严重死亡。 目前的治疗选择仅限于针对病毒株的疫苗接种，而且由于抗病毒药物的存在而存在问题 抵抗。迫切需要识别新的宿主先天免疫机制，以提供广泛的 预防流感的措施。支气管上皮细胞协调氧化的细胞外抗菌系统 存在于由蛋白质乳过氧化物酶(LPO)、硫氰酸盐阴离子(SCN-)组成的呼吸道表面液体中 )和过氧化氢(过氧化氢)。LPO利用过氧化氢将SCN-氧化成次硫氰酸盐(OSCN-)，已知在 体外抗病毒作用。双氧化酶1(Duox1)是一种在支气管上皮细胞中高表达的NADPH氧化酶， 是系统的过氧化氢来源。我们的长期目标是确定Duox1/H2O2/LPO/SCN- 抗病毒系统可用于治疗人类患者的流感感染。这个 本提案的目的是确定和表征Duox1和LPO的抗流感病毒作用。 在多个实验系统中。我们的初步数据显示：1)原代支气管上皮细胞失活 几种流感病毒以Duox1/H2O2/LPO/SCN依赖的方式存在，2)Duox1缺陷小鼠有 流感后死亡率和发病率增加，病毒清除和白细胞募集受损 体内感染，以及3)该机制的体外灭活流感作用可增强抑制 流感感染。基于这些数据，我们的中心假设是Duox1/H_2O_2/LPO/SCN-体系 在体外和体内都能减弱流感感染，并可增强抗流感作用。其基本原理是 建议的研究是，有必要更好地了解基于Duox1/LPO的抗病毒药物有多强大 系统是，以及如何将其用于治疗目的。主要假设将在细胞内进行检验-- 使用多种流感毒株的自由、呼吸道上皮和小鼠模型系统。预计 我们的目标将产生几个有影响力的结果，包括1)详细描述抗流感机制 Duox1/H_2O_2/LPO/SCN-系统的作用；2)测定Dux_1在体内与抗癌的相关性。 广泛的流感毒株；以及3)探索Duox1/H2O2/LPO/SCN-系统的治疗潜力 以改善流感的清除和减少相关的肺损伤。我们的创新工作表明， Duox1/H2O2/LPO/SCN-系统可灭活流感，并使用Duox1缺陷小鼠品系进行体内研究。 概述的工作的意义在于建立了一种新的先天免疫机制的相关性 哪些呼吸道可以加强以减轻流感感染或与流感一起使用 疫苗有可能提高疗效。总之，我们拟议的工作将在实地产生积极影响 通过鉴定Duox1和LPO作为新的， 支气管上皮对抗流感的重要武器。

Project summary description Influenza virus infections affect millions of people worldwide every year and cause serious mortality. Current treatment options are limited to viral strain-specific vaccination and are problematic due to antiviral drug resistance. There is an urgent need to identify novel host innate immune mechanisms providing broad range protection against influenza. Bronchial epithelial cells orchestrate an oxidative extracellular antimicrobial system present in the airway surface liquid consisting of the protein lactoperoxidase (LPO), the thiocyanate anion (SCN- ) and hydrogen peroxide (H2O2). LPO oxidizes SCN- using H2O2 into hypothiocyanite (OSCN-) that has known in vitro antiviral effects. Dual oxidase 1 (Duox1), an NADPH oxidase highly expressed in bronchial epithelial cells, is the H2O2 source for the system. Our long-term goal is to determine whether the Duox1/H2O2/LPO/SCN- antiviral system could be manipulated in influenza infection for therapeutic purposes in human patients. The objective of this proposal is to determine and characterize the antiviral role of Duox1 and LPO against influenza in multiple experimental systems. Our preliminary data show that 1) primary bronchial epithelial cells inactivate several influenza viruses in an Duox1/H2O2/LPO/SCN- -dependent manner, 2) Duox1-deficient mice have increased mortality and morbidity, impaired viral clearance and leukocyte recruitment following influenza infection in vivo, and 3) the in vitro influenza-inactivating effect of this mechanism can be enhanced to inhibit influenza infection. Based on these data, our central hypothesis is that the Duox1/H2O2/LPO/SCN- system attenuates influenza infection, both in vitro and in vivo, and can be boosted to fight influenza. The rationale for the proposed research is that there is a need to better understand how powerful the antiviral Duox1/LPO-based system is and how can it be manipulated for therapeutic purposes. The main hypothesis will be tested in cell- free, airway epithelial and mouse model systems using a wide range of influenza strains. It is anticipated that our aims will yield several impactful outcomes including 1) detailed description of the anti-influenza mechanism of action of the Duox1/H2O2/LPO/SCN- system; 2) determination of the in vivo relevance of Duox1 in fighting a wide range of influenza strains; and 3) exploring the therapeutic potential of the Duox1/H2O2/LPO/SCN- system to improve influenza clearance and to diminish associated lung damage. Our innovative work shows that the Duox1/H2O2/LPO/SCN- system inactivates influenza, and uses a Duox1-deficient mouse strain for in vivo studies. The significance of the outlined work relies in establishing the relevance of a novel innate immune mechanism of the airways that can be enhanced to attenuate influenza infections or applied in conjunction with influenza vaccines to potentially enhance efficacy. In summary, our proposed work will have a positive impact in the fields of airway epithelial biology and antiviral innate immune responses by identifying Duox1 and LPO, as novel, crucial weapons of the bronchial epithelium against influenza.