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Lymph node stromal cells coordinate immune cell environments during Aspergillus fumigatus infection

烟曲霉感染期间淋巴结基质细胞协调免疫细胞环境

基本信息

批准号：
10751936
负责人：
Cheryl Mai
金额：
$ 5.26万
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：
2023
资助国家：
美国
起止时间：
2023-07-01 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10751936
关键词：
ATAC-seq Adoptive Transfer Affect Allergic Bronchopulmonary Aspergillosis Anatomy Antifungal Agents Antigen-Presenting Cells Antigens Antimicrobial Resistance Aspergillosis Aspergillus Aspergillus fumigatus CCL20 gene CCR6 gene CD4 Positive T Lymphocytes CXCL10 gene CXCL9 gene CXCR3 gene Cell Communication Cell Differentiation process Cell physiology Cells Chronic Competence Critical Illness Cues Data Development Disease Dissection Dissociation Effector Cell Environment Epigenetic Process Exhibits Gene Targeting Genetic Goals Health Host Defense Human Image Analysis Immune Immune System Diseases Immune response Immunity Immunobiology Immunofluorescence Immunologic Immunofluorescence Microscopy Immunologic Deficiency Syndromes Infection Inflammatory Knowledge Life Ligands Lymphocyte Lymphoid Tissue Maps Mediastinal lymph node group Methods Microscopy Modification Molecular Mus Mycoses Neighborhoods Patients Pattern Physicians Population Positioning Attribute Primary Infection Process Property Proteins Resolution Respiratory Tract Infections Reticular Cell Role Scientist Signal Transduction Source Stromal Cells System T cell differentiation T cell response T-Lymphocyte T-Lymphocyte Subsets Testing Therapeutic Training Vaccines adaptive immune response adaptive immunity cell type chemokine chemokine receptor clinically relevant confocal imaging cytokine design dimensional analysis draining lymph node effector T cell experience human disease imaging approach immunomodulatory strategy improved outcome insight lymph node microenvironment lymph nodes member memory retention mortality mouse model novel novel strategies novel therapeutic intervention pathogen pathogenic fungus receptor response secondary lymphoid organ segregation single-cell RNA sequencing spatiotemporal therapy development tool trafficking transcriptomic profiling transcriptomics vaccine development

项目摘要

Project Summary The opportunistic fungal pathogen Aspergillus fumigatus presents a major health concern in immunodeficient and critically ill patients, with invasive disease contributing to high mortality rates. Infection with A. fumigatus elicits a diverse adaptive CD4 T cell response. Our preliminary data have demonstrated distinct TH1 and TH17 spatial neighborhoods within mediastinal lymph nodes (LNs) during A. fumigatus airway infection. This spatial organization positions activated CD4 T cells to receive tailored signals (e.g., antigens, cytokines) for optimal effector cell differentiation and function. Lymph node stromal cells (LNSCs) are known to provide guidance cues for immune cell trafficking during monotypic effector T cell responses. However, the mechanisms by which distinct TH1 and TH17 microenvironments are concurrently established within LNs remain unknown. Defining these mechanisms will offer insights into the functional importance of LN microenvironments in establishing human immunity to infection and inflammatory disorders for the identification of novel therapeutic strategies. Based upon our single-cell RNA-sequencing and flow cytometric analyses of FRC and effector T cell subsets during A. fumigatus challenge, we propose that (i) FRC subsets establish spatially distinct TH1 and TH17 neighborhoods by dynamically regulating chemotactic receptor-ligand axes, such as CXCR3:CXCL9/CXCL10 and CCR6:CCL20 and that ii) these neighborhoods can persist after pathogen clearance. We will address these hypotheses in a clinically relevant murine model of invasive pulmonary aspergillosis. In Specific Aim 1, we will characterize distinct spatiotemporal microenvironments formed by CD4 T cells and FRCs in A. fumigatus infection by applying spatial transcriptomic profiling followed by high-content immunofluorescence methods paired with a novel computational approach for image analysis. In Specific Aim 2, we will define the functional role of CD4 T cell spatial organization in A. fumigatus infection by perturbing FRC- dependent chemokine gradients in a cell type-dependent manner using FRC-specific gene targeting. We will also conduct adoptive co-transfers of A. fumigatus-specific CD4 T cells sufficient and deficient in CXCR3 and CCR6. In Specific Aim 3, we will investigate the durability of infection-driven spatial FRC diversity by studying epigenetic modifications in A. fumigatus-experienced FRCs. We will also employ high-content confocal imaging to determine whether prior infections affect FRC formation of T cell neighborhoods in subsequent infections. This study employs and develops novel genetic tools, microscopy methods, and computational approaches to generate a systems level understanding of secondary lymphoid organ immunobiology. Furthermore, this proposal is tailored for a physician-scientist in training as it investigates the mechanisms by which stromal cells 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烟曲霉提出了一个主要的健康问题，在免疫缺陷和重症患者，侵袭性疾病导致高死亡率。感染A.烟曲霉激发多样的适应性CD 4 T细胞应答。我们的初步数据表明，不同的TH 1和TH 17 纵隔淋巴结（LN）内的空间邻域在A.烟曲霉呼吸道感染这种空间组织定位活化的CD 4 T细胞以接收定制的信号（例如，抗原、细胞因子），效应细胞分化和功能。淋巴结基质细胞（LNSCs）是已知的提供指导线索在单型效应T细胞应答期间的免疫细胞运输。然而，不同的TH 1和TH 17微环境同时建立在LN内仍然未知。限定这些机制将提供深入了解LN微环境的功能重要性，人类对感染和炎性疾病的免疫力，用于鉴定新的治疗策略。基于我们对FRC和效应T细胞亚群的单细胞RNA测序和流式细胞术分析，在A. fumigatus挑战，我们建议（i）FRC亚群建立空间上不同的TH 1和TH 17 通过动态调节趋化性受体-配体轴，如 CXCR 3：CXCL 9/CXCL 10和CCR 6：CCL 20，并且ii）这些邻近区可以在病原体感染后持续存在，间隙我们将在临床相关的侵袭性肺动脉高压小鼠模型中阐述这些假设。曲霉病在具体目标1中，我们将描述CD 4形成的不同时空微环境， T细胞和FRC在A.通过应用空间转录组学分析，免疫荧光方法与图像分析的新计算方法配对。具体目标 2、明确CD 4 T细胞空间结构在A.通过干扰FRC- 使用FRC特异性基因靶向，以细胞类型依赖性的方式使用依赖性趋化因子梯度。我们将也进行A的收养共同转移。CXCR 3充足和缺乏的烟曲霉特异性CD 4 T细胞， CCR 6。在具体目标3中，我们将通过研究感染驱动的空间FRC多样性的持久性，表观遗传修饰A.经历过烟毒的FRC。我们还将采用高内涵的共聚焦成像以确定先前感染是否影响后续感染中T细胞邻域的FRC形成。这研究采用并开发了新型遗传工具、显微镜方法和计算方法，产生一个系统水平的次级淋巴器官免疫生物学的理解。而且这该提案是专为一个医生，科学家在培训，因为它调查的机制，基质细胞诱导对临床相关病原体A的适应性免疫。烟曲霉，与抗真菌的影响治疗策略和疫苗开发。