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: 7460678
• 负责人: EILEEN REMOLD-O'DONNELL
• 金额: $ 23.23万
• 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 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7460678
• 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）：流感是由流感病毒引起的一种传染性呼吸道疾病，在全球范围内造成了实质性疾病。治疗选择很少，大流行的威胁是恒定的。我们已经确定了一种内源性生物反应修饰蛋白SERPINB1，也称为MNEI（单核细胞中性粒细胞弹性酶抑制剂），可保护肺免受细菌感染的侵害，我们假设，我们可能会证明，可能有助于保护侵害流感。使用肺部感染的啮齿动物模型，我们表明SERPINB1/MNEI可以保护先天的抗细菌宿主防御，并防止炎症和肺损伤。如预期的那样，SERPINB1/MNEI在体外没有抗细菌作用，因为其生化功能是选择性地有效抑制中性粒细胞丝氨酸蛋白酶（NSP）。 NSP（弹性酶，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，它们在炎性肺病中大量释放，并且是有力的病理学剂，是炎症性趋化因子过量产生的强大病理学剂，诱导抗肺动物造成抗摩尔抗性的肺部组织造成直接的蛋白水解损伤。 NSP靶向并受到MNEI保护的宿主防御分子中的突出是肺表面活性剂蛋白-D（SP-D），它中和，中和许多流感病毒菌株并增强了清除率，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）项目将检验以下假设：中性粒细胞丝氨酸蛋白酶（NSP）有助于MNEI/SERPINB1预防流感的致病性。我们将使用通过删除SERPINB1/MNEI基因在实验室中产生的NSP活性增加的小鼠模型，并将确定这些小鼠在感染流感病毒菌株（IAV）时是否显示出增加的发病率（体重下降）和死亡率。我们将通过测量病毒滴度，肺损伤，炎症细胞因子，肺泡巨噬细胞的流入和中性粒细胞的流入，中性粒细胞的坏死，中性粒细胞和Intact sp-D和Nspspclated Intactived-D-D-D-D-D-D-D-D-D-D-D-D-D-D-D-D-D-D-D-D-D-D-D-D-D-D-D-D-D-D-D-spspecivived-clativevivev。最后，我们将测试重组MNEI是否挽救了MNEI - / - 小鼠的有缺陷反应，并增强了野生型小鼠对流感病毒的抗病毒宿主反应。成功识别NSP作为流感中致病性的介体将是迈向一种新型治疗的一步。作为流感的治疗性，NSP的抑制剂，尤其是MNEI，一种天然存在的生物反应修饰剂，将在机械上独立于当前药物，因此适用于组合使用。由于MNEI通过保护先天宿主的免疫力来发挥作用，因此预计有疗效对具有不良适应性免疫的人（包括非常年轻且非常老）。重要的是，MNEI不需要结合病毒决定因素，因此不会预期抗性的发展。流感是一种由流感病毒引起的一种传染性呼吸道疾病，是导致实质性疾病的原因，并且在全球范围内真正威胁出来的剧烈菌株。我们已经确定了内源性生物反应修饰蛋白的MNEI（单核细胞/中性粒细胞弹性酶抑制剂），可保护肺部免受细菌感染。在拟议的R21（探索/ - 发展赠款）项目中，我们将使用实验室中开发的小鼠模型来检验以下假设：MNEI将被证明可用于保护流感。