Dendritic cell heterogeneity and its role in pulmonary fungal infection

树突状细胞异质性及其在肺部真菌感染中的作用

基本信息

批准号：
10327607
负责人：
Deeksha Deep
金额：
$ 4.9万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-12-14 至 2024-12-13
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10327607
关键词：
Ablation Acute Adoptive Transfer Allergic Bronchopulmonary Aspergillosis Anatomy Anti-Inflammatory Agents Antifungal Agents Antifungal Therapy Antigen Presentation Antigen-Presenting Cells Antigens Aspergillus fumigatus CD4 Positive T Lymphocytes CD8B1 gene Cells Chronic Clinical Complex Computing Methodologies Critical Illness Cytometry Data Dendritic Cells Exhibits Flow Cytometry Genetic Genetic Transcription Goals Health Helper-Inducer T-Lymphocyte Heterogeneity Human ITGAM gene Immune Immune response Immunity Immunobiology Immunocompromised Host Immunofluorescence Immunologic In Vitro Infection Inflammation Inflammatory Knowledge Label Lung MHC Class II Genes Mediastinal lymph node group Mediator of activation protein Methods Microscopy Mus Mycoses Natural Immunity Pathogenicity Pathologic Processes Patients Physicians Physiological Population Positioning Attribute Property Regulation Regulatory T-Lymphocyte Role Scientist Spatial Distribution Study models Syndrome System T cell response T-Cell Activation T-Lymphocyte T-bet protein Testing Therapeutic Therapeutic Intervention Training Vaccines XCR1 gene adaptive immune response adaptive immunity antigen-specific T cells attributable mortality base clinically relevant differential expression draining lymph node human disease in vivo mortality mouse model novel novel strategies novel therapeutic intervention pathogen pathogenic fungus patient population response single cell analysis stem therapy development tool trafficking transcriptome sequencing uptake vaccine development

项目摘要

Project Summary The opportunistic fungal pathogen Aspergillus fumigatus is a major health concern in immunocompromised and critically ill patients, manifesting as a variety of pulmonary conditions ranging from acute to chronic. In these clinical syndromes, A. fumigatus elicits a diverse adaptive CD4 T cell response, but the mechanisms by which these varied responses are induced remains unknown. Conventional dendritic cells (cDCs) are crucial for sensing and initiating immune responses to this fungal pathogen and are the likely mediators of diverse T cell responses to A. fumigatus. Recent studies have established the transcriptional basis for cDC heterogeneity through single cell analyses, specifically demonstrating a division in the cDC2 population, which are considered the canonical antigen presenting cells, expressing MHC Class II and priming CD4 T cells. We now separate cDC2s into two novel subsets—cDC2A and cDC2B—based on differential expression of the transcription factors T-bet and RORγt, respectively. In addition, the anatomic positioning of cDC2s is known to facilitate spatial colocalization with pathogen-derived products allowing for efficient pathogen sensing, antigen uptake, and subsequent CD4 T cell activation. Based on the pathogenic properties of A. fumigatus and recent discoveries of cDC2 heterogeneity, our specific hypothesis is that cDC2A and cDC2B subsets will localize differently in lung and draining lymph node and facilitate different adaptive CD4 T cell response types to A. fumigatus. In this study, we investigate cDC2 subsets’ functionality in a clinically relevant murine model of invasive pulmonary aspergillosis. In Specific Aim 1, we will define the temporal and spatial dynamics of cDC2 subsets during acute A. fumigatus infection by applying high-content immunofluorescence methods and flow cytometry. We will also assess cDC2A and cDC2B functional properties in vitro. In Specific Aim 2, we will elucidate the direct impact of cDC2 subsets on adaptive immunity in A. fumigatus infection by implementing genetic tools to specifically ablate cDC2A and cDC2B subsets and establish their functional significance in vivo. In addition, by investigating spatial reorganization of other immune cell subsets in subset-specific ablation of cDC2A and cDC2B, we can identify their significance in cellular circuits governing lung immunity. This study employs and develops novel genetic tools, microscopy methods, and computational approaches to generate a systems level understanding of lung immunobiology and study host-pathogen interactions. Furthermore, this proposal is tailored for a physician-scientist in training, as it investigates the basic features of and mechanisms by which cDC2 subsets induce adaptive immunity to the clinically relevant pathogen A. fumigatus, with implications for anti-fungal therapeutic strategies and vaccine development.

项目摘要机会性真菌病原体曲霉菌富瓜曲霉是免疫功能低下的主要健康问题，重症患者，表现为从急性到慢性的多种肺部疾病。在这些临床综合征，A。fumigatus引起了潜水员的自适应CD4 T细胞反应，但其机制这些多样的反应是未知的。常规的树突状细胞（CDC）对于感应并开始对这种真菌病原体的免疫反应，并且可能是潜水员T细胞的介体对烟曲霉的反应。最近的研究确定了CDC异质性的转录基础通过单细胞分析，专门证明了Cdc2种群中的分裂，这被认为表达MHC II类和启动CD4 T细胞的规范抗原呈递细胞。我们现在分开 Cdc2分为两个新的子集-CDC2A和CDC2B，基于转录因子的差异表达 T-BET和RORγT。此外，已知Cdc2s的解剖定位可促进空间与病原体衍生的产品共定位，允许有效的病原体传感，抗原摄取和随后的CD4 T细胞激活。基于烟曲霉的致病特性以及最近发现的发现 CDC2异质性，我们的具体假设是CDC2A和CDC2B子集将以不同的位置在肺和排水淋巴结，并促进对烟曲霉的不同自适应CD4 T细胞反应类型。在这项研究中，我们研究了CDC2子集在临床相关的侵入性鼠模型中的功能肺曲霉病。在特定目标1中，我们将定义CDC2子集的临时和空间动力学在急性烟曲霉感染期间，通过应用高内核免疫荧光方法和流式细胞仪。我们还将在体外评估CDC2A和CDC2B功能特性。在特定目标2中，我们将阐明 Cdc2子集对烟曲霉感染中自适应免疫的直接影响通过实施遗传工具特别是烧蚀cdc2a和cdc2b子集，并在体内建立其功能意义。另外，由调查其他免疫细胞亚群的空间重组在CDC2A和 CDC2B，我们可以确定它们在控制肺免疫组织化学的细胞回路中的重要性。本研究员工和开发新型的遗传工具，显微镜方法和计算方法来生成系统级别了解肺免疫生物学和研究宿主 - 病原体相互作用。此外，该建议是为培训的身体科学家量身定制，因为它研究了该机制的基本特征和机制 CDC2子集诱导适应性免疫学对临床相关的病原体烟曲霉，对抗真菌治疗策略和疫苗开发。