Systems Analysis of Innate Immune Responses to Fungal-Derived Carbohydrates

对真菌衍生碳水化合物的先天免疫反应的系统分析

• 批准号: 8970198
• 负责人: Jatin M Vyas
• 金额: $ 26.1万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8970198
• 关键词: Antifungal Agents; Aspergillosis; Aspergillus fumigatus; Bioinformatics; Biological Assay; Breathing; Carbohydrates; Cell Wall; Cells; Chitin; Clinical; Collection; Development; Diagnosis; Distal; Environment; Global Change; Glucans; Hematopoietic stem cells; Host Defense; Human; Immune; Immune response; Immune system; Immunocompromised Host; Immunologics; Industrial fungicide; Infection; Inflammatory; KAI1 gene; Knowledge; LAMP-1; Lead; Lectin Receptors; Life; Ligands; Ligation; Lung; MHC Class II Genes; Mannans; Mass Spectrum Analysis; Mediating; Membrane; Membrane Proteins; Molds; Mycoses; Organ Transplantation; Organism; Outcome; Output; Pathway Analysis; Patients; Pattern; Phagocytosis; Phagosomes; Pharmaceutical Preparations; Play; Polysaccharides; Polystyrenes; Prevention; Proteins; Proteome; Proteomics; Receptor Signaling; Recovery; Recruitment Activity; Reproduction spores; Role; Shapes; Signal Pathway; Signal Transduction; Solid; Stem cell transplant; Systems Analysis; TLR2 gene; TLR4 gene; Therapeutic; Time; Tumor Necrosis Factor-alpha; Work; adaptive immunity; asexual; candidemia; comparative; crosslink; cytokine; dectin 1; fungus; galactomannan; in vitro activity; insight; interest; macrophage; monocyte; multidisciplinary; novel; outcome forecast; particle; pathogen; public health relevance; receptor; response; uptake; vaccine development

 DESCRIPTION (provided by applicant): Aspergillus fumigatus is a saprophytic filamentous fungus whose asexual spores called conidia are widespread in the environment, acquired through inhalation and small enough to reach the distal airways. Approximately 10% of patients with hematopoietic stem cell transplants or solid organ transplants will develop invasive aspergillosis, a life-threatening infection. Despite the development of effective fungicidal agents the prognosis for disseminated infection is quite poor, indicating that knowledge of the rules that govern the host defense against A. fumigatus is critical for development of new prevention and therapeutic strategies. Evidence for the importance of innate immune mechanisms in fungal defense is mounting. The fungal ß-1,3 glucan receptor Dectin-1 is essential in pulmonary defense against A. fumigatus and collaborates with TLR2 and TLR4 in providing critical immune signals that coordinate cytokine secretion and development of the adaptive immunity. The fungal cell wall serves as the initial point of contact in the pathogen-host relationship. Despite elegant ultra-structural studies on the topology of the fungal cell wall, our understanding of the polysaccharide constituents that interact with mammalian cellular receptors is poorly understood. ß-1,3 glucan, a proinflammatory polysaccharide found on numerous pathogenic fungi, is the best studied fungal carbohydrate. The precise response triggered by other fungal carbohydrates including chitin, mannan, galactoaminogalactan, galactomannan and α-1,3 glucan remains incompletely understood. In order to understand better the contribution of these fungal polysaccharides to the innate immune response, we have made the following key observations that are the rationale for our proposed work: 1) when present in phagosomes, fungi trigger specific recruitment of multiple proteins of immunologic interest including CD63, CD82, class II MHC, LC3, LAMP-1 and TLR9. In sharp contrast, polystyrene beads failed to do so. 2) we generated size-matched polystyrene beads that possess a uniform coat of a single fungal-derived carbohydrate covalently attached to probe the immune response. These fungal like particles are biologically active and trigger cytokine responses 3) using fungal like particle, we generated phagosomes in macrophages and determined that the phagosomal proteome by mass spectrometry. We hypothesize that the content of the phagosome triggers the assembly of specific mammalian proteins to this compartment and that this assembly of proteins orchestrates the net immunologic output from this cell. We propose to: 1] Determine the mammalian proteins recruited to the phagosome containing different fungal like particles and 2] Profile the cytokine signature in human monocytes to fungal-like particles. We will apply advanced proteomics and multiplex cytokine assay to achieve these aims. Knowledge gained regarding the mechanism of carbohydrate recognition, its influence on the phagosome formation and cytokine signature will be important in furthering our understanding of the innate immune response to A. fumigatus, and could lead to novel insights to vaccine development against this deadly pathogen.

 描述（由适用提供）：曲霉菌是一种腐生的丝状菌丝真菌，其无性孢子被称为分生孢子在环境中宽度，通过吸入而获得，足够小，可以到达远端气道。大约10％的造血干细胞移植或固体器官移植的患者会出现侵入性的曲霉病，这是一种威胁生命的感染。尽管发展了有效的杀真菌剂，但对传播感染的预后还是很差的，这表明对规则的了解 对烟曲霉的辩护负责，对于开发新的预防和理论策略至关重要。先天性免疫组机制在真菌防御中的重要性的证据正在加剧。真菌ß-1,3葡聚糖受体Dectin-1对于针对烟曲霉的肺防御至关重要，并与TLR2和TLR4合作提供了关键的免疫化信号，以协调适应性免疫的细胞因子分泌和发展。真菌细胞壁是病原体宿主关系中接触的初始点。尽管对真菌细胞壁拓扑的优雅超结构研究，但我们对与哺乳动物细胞接收器相互作用的多糖构成的理解知之甚少。 ß-1,3葡聚糖是在许多致病真菌上发现的一种促炎多糖，是最好的研究生真菌碳氢化物。其他真菌碳水化合物触发的精确反应，包括几丁质，曼南，半乳糖氨基乳糖，半乳糖量和α-1,3葡聚糖，仍然不完全理解。为了更好地理解这些真菌多糖对先天免疫的贡献，我们进行了以下主要观察结果，这些观察结果是我们提出的工作的基本原理：1）当在吞噬体中存在，真菌会触发特定的免疫兴趣的特定蛋白质，包括免疫兴趣的多种蛋白质，包括CD63，CD82，II MHC，II MHC，LC3，LC3，LC3，LAMP，LAMP和TLR。鲜明的对比，聚苯乙烯珠没有这样做。 2）我们产生了尺寸匹配的聚苯乙烯珠，该珠具有共价附着的单个真菌衍生的碳水合物的均匀涂层，以探测免疫增强件。这些真菌像颗粒具有生物活性，并触发细胞因子反应3）使用真菌颗粒，我们在巨噬细胞中产生了吞噬体，并确定通过质谱法进行吞噬体蛋白质组。我们假设吞噬体的含量会触发特定的哺乳动物蛋白的组装到该区室，并且这种蛋白质组装策划了该细胞的净免疫输出。我们建议：1]确定募集到含有不同真菌（如颗粒）的吞噬体的哺乳动物蛋白，并介绍2]人类单核细胞中的细胞因子特征到真菌样颗粒。我们将应用晚期蛋白质组学和多种细胞因子测定来实现这些目标。关于碳水性识别机制的知识，其对吞噬体形成和细胞因子特征的影响对于进一步的理解对我们对烟曲霉的先天免疫反应至关重要，并可能导致对这种致命的病原体的疫苗发育的新见解。