SERPINB1/MNEI: Role in innate immune defense against influenza virus infection

SERPINB1/MNEI：在针对流感病毒感染的先天免疫防御中的作用

• 批准号: 7304824
• 负责人: EILEEN REMOLD-O'DONNELL
• 金额: $ 28.41万
• 依托单位: IMMUNE DISEASE INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-15 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7304824
• 关键词: Acute; Alveolar; Alveolar Macrophages; Anti-Bacterial Agents; Anti-Inflammatory Agents; Anti-inflammatory; Antiviral Agents; Bacterial Infections; Binding; Biochemical; Biological Response Modifiers; Body Weight decreased; Bronchoalveolar Lavage Fluid; C57BL/6 Mouse; Cathepsin G; Cells; Cessation of life; Chemicals; Chronic; Cleaved cell; Collagen; Condition; Cytoplasmic Granules; Detection; Development; Dose; Edema; Effectiveness; Elastases; Endopeptidases; Enzyme-Linked Immunosorbent Assay; Enzymes; Epidemic; Epithelial Cells; Exploratory/Developmental Grant; Exposure to; Fostering; Gene Deletion; Generations; Genes; Genotype; Granulocyte-Macrophage Colony-Stimulating Factor; Hand; Hemagglutinin; Hospitalization; Host Defense; Human; IL6 gene; Immune; Immune response; Immunity; In Vitro; Individual; Infection; Inflammation; Inflammatory; Influenza; Influenza A virus; Influenza Therapeutic; Injury; Interferons; Interleukin-1; Interleukin-10; Laboratories; Lectin; Life; Link; Liquid substance; Lower respiratory tract structure; Lung; Lung Inflammation; Lung Lavage Fluid; Lung diseases; Measures; Mediating; Mediator of activation protein; Modeling; Monitor; Morbidity - disease rate; Mus; Necrosis; Neuraminidase; Neutrophil Infiltration; Pancreatic Elastase; Pathogenicity; Pathologist; Peptide Hydrolases; Permeability; Peroxidase; Phagocytes; Pharmaceutical Preparations; Pilot Projects; Prevention; Proteinase 3; Proteins; Pulmonary Surfactant-Associated Protein D; RNA; Rattus; Recombinants; Recruitment Activity; Resistance; Resistance development; Reverse Transcription; Rodent Model; Role; Serine Protease; Structure of parenchyma of lung; TNF gene; Testing; Therapeutic; Time; Upper respiratory tract; Vaccines; Viral; Virulent; Virus; Virus Diseases; Weight; Western Blotting; Wild Type Mouse; anti-influenza; antimicrobial; chemokine; clinical efficacy; cytokine; day; env Gene Products; experience; extracellular; glycosylation; in vivo; influenza virus strain; influenzavirus; inhibitor/antagonist; innovation; killings; lung injury; macrophage; monocyte; mortality; mouse model; neutrophil; neutrophil elastase inhibitor; novel; novel therapeutics; pandemic disease; pathogen; prevent; protective effect; receptor; respiratory; response; therapeutic target

DESCRIPTION (provided by applicant): Influenza, a contagious respiratory illness caused by influenza virus, is responsible for substantial illness worldwide. Treatment options are few and the threat of a pandemic is constant. We have identified SERPINB1, an endogenous biological response modifier protein, also called MNEI (monocyte neutrophil elastase inhibitor) that protects the lung against bacterial infection and that, we hypothesize, may prove useful in protecting against influenza. Using rodent models of pulmonary infection, we showed that SERPINB1/MNEI protects innate anti-bacterial host defense and prevents inflammation and lung injury. SERPINB1/MNEI had no anti-bacterial effect in vitro, as anticipated, since its biochemical function is to selectively and efficiently inhibit neutrophil serine proteases (NSPs). The NSPs (elastase, cathepsin G and proteinase-3) are active proteases stored in neutrophil granules, which are released in large amounts in inflammatory lung disease and are powerful pathological agents that induce overproduction of inflammatory chemokines, direct proteolytic injury to lung tissue and loss of anti-microbial defense by degrading innate protective molecules. Prominent among the host defense molecules targeted by NSPs and protected by MNEI is pulmonary surfactant protein-D (SP-D), which neutralizes many influenza virus strains and enhances their clearance and protects host defense by anti- inflammatory action on alveolar macrophages. The proposed R21 (pilot) project will test the hypothesis that neutrophil serine proteases (NSPs) contribute to, and MNEI/SERPINB1 protects against, pathogenicity of influenza. We will use a mouse model of increased NSP activity recently generated in our laboratory by deletion of the SERPINB1/MNEI gene and will determine whether these mice show increased morbidity (loss of body weight) and mortality on infection with strains of influenza A virus (IAV). We will test for abnormalities of the innate immune response (early host response) of IAV infected mnei-/- mice by measuring viral titer, lung injury, inflammatory cytokines, influx of alveolar macrophages and neutrophils, necrosis of neutrophils, and levels of intact SP-D and NSP-cleaved inactivated SP-D. Finally, we will test whether recombinant MNEI rescues the defective response of mnei-/- mice and enhances the anti-viral host response of wild-type mice to influenza virus. Successful identification of NSPs as mediators of pathogenicity in influenza would be a step toward a novel therapeutic. As a therapeutic for influenza, inhibitors of NSPs and especially MNEI, a naturally occurring biological response modifier, would be mechanistically independent of current drugs and therefore suitable for combination use. Because MNEI functions by protecting innate host immunity, efficacy is anticipated for individuals with deficient adaptive immunity including the very young and very old. Importantly, MNEI has no need for binding viral determinants, and thus development of resistance is not anticipated. Influenza, a contagious respiratory illness caused by influenza virus, is responsible for substantial illness, and there is real threat of worldwide spread of emerging more virulent strains. We have identified MNEI (monocyte/neutrophil elastase inhibitor), an endogenous biological response modifier protein, that protects the lung against bacterial infection. In the proposed R21 (Exploratory/- Developmental Grant) project, we will use a mouse model developed in our laboratory to test the hypothesis that MNEI will prove useful in protecting against influenza.

描述（由申请人提供）： 流感是一种由流感病毒引起的传染性呼吸道疾病，是全世界重大疾病的原因。治疗选择很少，而且大流行的威胁始终存在。我们已经鉴定出 SERPINB1，一种内源性生物反应调节蛋白，也称为 MNEI（单核细胞中性粒细胞弹性蛋白酶抑制剂），可以保护肺部免受细菌感染，我们假设它可能有助于预防流感。使用肺部感染的啮齿动物模型，我们发现 SERPINB1/MNEI 可以保护宿主先天抗菌防御并预防炎症和肺损伤。正如预期的那样，SERPINB1/MNEI 在体外没有抗菌作用，因为其生化功能是选择性且有效地抑制中性粒细胞丝氨酸蛋白酶（NSP）。 NSP（弹性蛋白酶、组织蛋白酶 G 和蛋白酶-3）是储存在中性粒细胞颗粒中的活性蛋白酶，在炎症性肺部疾病中大量释放，是强大的病理因子，可诱导炎症趋化因子过量产生，直接对肺组织造成蛋白水解损伤，并通过降解先天保护分子而丧失抗微生物防御能力。 NSP 靶向并受 MNEI 保护的宿主防御分子中最突出的是肺表面活性蛋白-D (SP-D)，它可以中和许多流感病毒株并增强其清除率，并通过对肺泡巨噬细胞的抗炎作用来保护宿主防御。拟议的 R21（试点）项目将测试以下假设：中性粒细胞丝氨酸蛋白酶 (NSP) 有助于流感的致病性，而 MNEI/SERPINB1 可以预防流感的致病性。我们将使用我们实验室最近通过删除 SERPINB1/MNEI 基因而产生的 NSP 活性增加的小鼠模型，并将确定这些小鼠感染甲型流感病毒 (IAV) 株后是否表现出发病率（体重减轻）和死亡率增加。我们将通过测量病毒滴度、肺损伤、炎症细胞因子、肺泡巨噬细胞和中性粒细胞的流入、中性粒细胞坏死以及完整 SP-D 和 NSP 裂解的灭活 SP-D 的水平来测试 IAV 感染的 mnei-/- 小鼠先天免疫反应（早期宿主反应）的异常。最后，我们将测试重组MNEI是否可以挽救mnei-/-小鼠的缺陷反应并增强野生型小鼠对流感病毒的抗病毒宿主反应。成功鉴定 NSP 作为流感致病性介质将是迈向新型治疗方法的一步。作为流感的治疗剂，NSP 抑制剂，特别是 MNEI（一种天然存在的生物反应调节剂）在机制上独立于现有药物，因此适合联合使用。由于 MNEI 通过保护先天宿主免疫发挥作用，因此预计对适应性免疫缺陷的个体（包括非常年轻和非常年老的人）有效。重要的是，MNEI 不需要结合病毒决定簇，因此预计不会产生耐药性。流感是一种由流感病毒引起的传染性呼吸道疾病，是造成严重疾病的原因，而且新出现的毒性更强的毒株确实存在全球传播的威胁。我们已经鉴定出 MNEI（单核细胞/中性粒细胞弹性蛋白酶抑制剂），一种内源性生物反应调节蛋白，可以保护肺部免受细菌感染。在拟议的 R21（探索性/-发展资助）项目中，我们将使用我们实验室开发的小鼠模型来测试 MNEI 将证明在预防流感方面有用的假设。