Defining the role of mast cells during influenza A virus infection

定义肥大细胞在甲型流感病毒感染过程中的作用

• 批准号: 8225422
• 负责人: JOSHUA J OBAR
• 金额: $ 10.8万
• 依托单位: MONTANA STATE UNIVERSITY - BOZEMAN
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-15 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8225422
• 关键词: Address; Age; Alveolar; Area; Bacterial Infections; Biological Assay; Bone Marrow; Bronchi; C-KIT Gene; Cells; Chimera organism; Chymase; Color; Data; Dendritic Cells; Detection; Disease Outbreaks; Disease Outcome; Environment; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Event; Faculty; Family; Flow Cytometry; Foundations; Frequencies; Future; Gastrointestinal tract structure; Genitourinary system; Hematopoiesis; Immune; Immune response; Infection; Inflammation; Inflammatory; Inflammatory Response; Influenza A Virus, H1N1 Subtype; Influenza A Virus, H5N1 Subtype; Influenza A virus; Institutes; Invaded; Knowledge; Lung; Mediating; Morbidity - disease rate; Mus; Mutation; Parasitic infection; Pathogenicity; Pathologic; Pathology; Pathway interactions; Peptide Hydrolases; Pharmacologic Substance; Plaque Assay; Play; Population; Primary Cell Cultures; Principal Investigator; Proteinase-Activated Receptors; Proto-Oncogene Protein c-kit; Public Health; Regulation; Research; Respiratory Tract Infections; Response Elements; Role; Running; Severities; Skin; Staging; System; Techniques; Testing; Time; Training; Tryptase; Universities; Viral; Virus Diseases; Work; cell type; chemokine; cytokine; experience; flexibility; immunopathology; interest; lung injury; mast cell; member; mortality; new therapeutic target; novel; pandemic disease; pathogen; professor; programs; reconstitution; respiratory; response; seasonal influenza; sound

DESCRIPTION (provided by applicant): Influenza A virus (IAV) and especially the highly pathogenic IAV strains, such as those causing the recent H5N1 and H1N1 outbreaks, represent a significant public health concern. These strains of IAV cause significantly greater morbidity and mortality than seasonal IAV strains. However, the exact cause of the increased morbidity and mortality remains controversial, with increased viral pathogenicity, increased immunopathology, and increase co-infections proposed as possible mechanisms. This proposal will examine the early host response elements to IAV infection, specifically examining the role that mast cells and protease- activated receptors play in regulating the morbidity and mortality associated with IAV infection. Mast cells could be critical in orchestrating the host response to respiratory infection with IAV, because they are found at elevated frequencies around the bronchi within the lungs. Furthermore, our preliminary data show that mast cell-deficient mice develop significantly less IAV-association morbidity. Therefore, we propose to determine how mast cells are activated during respiratory infection with IAV and what role they play in regulating disease outcome. Secondly, we will determine how mast cells are orchestrating the deleterious host inflammatory response following IAV infection. Specifically, mast cell activation of the protease-activated receptor family on alveolar epithelial cells will be explored, as well as the subsequent regulation of inflammatory chemokines and cytokines by both mast cells and the protease-activated receptor family. Our preliminary data support the hypothesis that mast cells and the protease-activated receptor family are important in regulating morbidity associated with respiratory IAV infection. Thus, by understanding these early events, we hope to identify novel pharmacological targets, which can be used to limit morbidity and mortality from IAV infection. As can be seen, the focus of this proposal is quite distinct from the previous research that I have conducted. However, I have been well trained in the experimental systems and techniques needed for the successful completion of the proposed studies. For example, I have gained extensive experimental experience in the use of mice for understanding the immune response to numerous viral pathogens, including IAV. In addition, I have gained substantial training in many of the techniques required for the successful completion of the proposed studies, which include multi-parameter flow cytometry (we are routinely conducting 12 parameter studies now), primary cell culture from bone marrow, Luminex" bead assays for the detection of multiple cytokines/chemokines at the same time, viral plaque assays, adoptive cellular transfers, and ELISA assays. Together, this previous experience provides a strong experimental foundation, which will allow me the flexibility to apply this knowledge and technical ability to a distinct area of study, therefore developing a new and independent research direction that addresses critical gaps in knowledge of immune regulation of viral infection as an Assistant Professor. Furthermore, my current and future research institutes provide me ample technical opportunities to conduct these studies, including access to a 10-color BD LSRII cytometer, a g-cell irradiator for making bone marrow chimeras, and BioPlex200 machine for running Luminex plates. Additionally, faculty members at both my current and future universities provide a sound network of knowledge which will be invaluable for the successfully completion of the proposed studies. Each year, influenza A virus infection leads to significant morbidity and mortality within certain groups, while periodic pandemic strains cause significantly greater morbidity and mortality throughout the entire population. What causes the significantly elevated rates of morbidity and mortality remains controversial. This proposal will examine the early events during the host response to influenza A virus infection and could elucidate potential novel therapeutic targets for limiting morbidity and mortality associated with influenza A virus infection.

描述（由申请人提供）：流感病毒（IAV），尤其是高度致病的IAV菌株，例如引起最近H5N1和H1N1爆发的菌株，这是一个重大的公共卫生问题。与季节性IAV菌株相比，这些IAV菌株会导致发病率和死亡率明显更高。但是，发病率和死亡率增加的确切原因仍然引起争议，病毒性致病性增加，免疫病理学增加并增加了提出的共同感染作为可能的机制。该建议将检查对IAV感染的早期宿主反应元件，特别研究了肥大细胞和蛋白酶活化受体在调节与IAV感染相关的发病率和死亡率中起作用的作用。肥大细胞对于策划宿主对呼吸道感染的反应至关重要，因为它们是在肺部支气管周围较高的频率下发现的。此外，我们的初步数据表明，肥大细胞缺陷型小鼠的发病率明显降低。因此，我们建议确定在使用IAV呼吸道感染期间如何激活肥大细胞，以及它们在调节疾病结果中的作用。其次，我们将确定肥大细胞如何在IAV感染后如何策划有害的宿主炎症反应。具体而言，将探索蛋白酶激活的受体家族在肺泡上皮细胞上的肥大细胞活化，以及肥大细胞和蛋白酶激活受体家族对炎症趋化因子和细胞因子的随后调节。我们的初步数据支持以下假设：肥大细胞和蛋白酶激活的受体家族对于调节与呼吸IAV感染相关的发病率很重要。因此，通过了解这些早期事件，我们希望确定新型的药理学靶标，这些靶标可用于限制IAV感染中的发病率和死亡率。 可以看出，该提案的重点与我先前的研究完全不同。但是，在成功完成拟议的研究所需的实验系统和技术中，我接受了良好的培训。例如，我在使用小鼠来理解包括IAV在内的众多病毒病原体的免疫反应方面获得了丰富的实验经验。此外，我在成功完成拟议的研究所需的许多技术中都获得了实质性培训，其中包括多参数流式细胞仪（我们现在常规进行12个参数研究），来自骨髓的原代细胞培养，luminex”珠子“珠子”，用于检测多个细胞因子/趋化因子，以下情况下，该分别是这种情况，这是该时间，病毒pla pla sass and and and and and。 previous experience provides a strong experimental foundation, which will allow me the flexibility to apply this knowledge and technical ability to a distinct area of​​ study, therefore developing a new and independent research direction that addresses critical gaps in knowledge of immune regulation of viral infection as an Assistant Professor. Furthermore, my current and future research institutes provide me ample technical opportunities to conduct these studies, including access to a 10-color BD LSRII cytometer, a g-cell irradiator for making bone marrow用于运行Luminex板的嵌合体和Bioplex200机器。此外，我目前和未来的大学的教职员工提供了一个合理的知识网络，这对于成功完成拟议的研究将是无价的。 每年，流感的一种病毒感染会导致某些组的显着发病率和死亡率，而周期性的大流行菌株在整个人群中会导致发病率和死亡率明显更高。导致发病率和死亡率显着提高的原因仍然存在争议。该提案将检查宿主对流感病毒感染的宿主反应期间的早期事件，并可以阐明潜在的新型治疗靶标，以限制与流感A的病毒感染相关的发病率和死亡率。