Tracing spatial organization of germinal centers in rhesus macaques

追踪恒河猴生发中心的空间组织

• 批准号: 10762072
• 负责人: Ahmet F. Coskun
• 金额: $ 31.64万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10762072
• 关键词: AIDS diagnosis; Acquired Immunodeficiency Syndrome; Animals; Antibodies; B-Cell Activation; B-Cell Development; B-Lymphocyte Subsets; B-Lymphocytes; Bioinformatics; CD4 Positive T Lymphocytes; Cell Communication; Cell Maturation; Cells; Coupled; Cytokine Gene; Defect; Disease Progression; Emerging Technologies; Evolution; Extracellular Matrix; Follicular Dendritic Cells; Gene Expression; Gene Expression Profiling; Genes; Goals; HIV; Human; Image; Immune; Immune response; Immune system; Immunity; Immunology; Impairment; Individual; Infection; Inflammation; Interleukin-10; Investigation; Lymph Node Tissue; Lymphoid Cell; Lymphoid Tissue; Macaca; Macaca mulatta; Maps; Memory B-Lymphocyte; Methods; Modeling; Molecular; Mutate; Pathogenesis; Pathology; Pathway interactions; Patients; Persons; Phenotype; Plasma Cells; Proteomics; Reaction; Regulation; Research; Research Personnel; Resolution; Role; SIV; Site; Spatial Distribution; Specialized Center; Structure; Structure of germinal center of lymph node; T cell regulation; T-Lymphocyte; Techniques; Technology; Testing; Tissues; Up-Regulation; Vaccinated; Viral; Virus; Virus Diseases; antiretroviral therapy; cell type; cytokine; experience; follow-up; high reward; high risk; human subject; human tissue; immune activation; immunoregulation; innovation; insight; interdisciplinary approach; lymph nodes; mortality; multiple omics; nonhuman primate; peripheral blood; pre-clinical; protein distribution; response; secondary lymphoid organ; simian human immunodeficiency virus; spatiotemporal; transcriptomics; transmission process; viral DNA

Germinal centers (GCs) are the microstructural sites in secondary lymphoid organs, but GC structures are impaired in AIDS patients due to the loss of supporting CD4+ T cells, leading the deficiencies in immune responses and even inefficacies in antiretroviral therapies in HIV-infected individuals. To solve this challenge, rhesus macaques are well-established nonhuman primate models (NHPs) to study the immunopathogenesis of HIV. However, the spatial coordination of Follicular dendritic cells (FDCs), T cells, B cells, extracellular matrix (ECM), and cytokine regulation of simian immunodeficiency virus (SIV)+ infected macaques and SIV- macaques is still not clearly understood. Thus, there is a critical need to decipher the spatial and temporal control of (1) cytokine, (2) ECM, and (3) T-B cell interactions at the single cell level in GCs for identifying the (1) spontaneous activation and (2) SIV-infection-induced responses in the immune system of HIV/SIV disease. To shed light on immune regulation in lymph node tissues of SIV+ and SIV- macaques and human donors, this project will leverage spatial proteomic and transcriptional profiling to map B cell subsets and their interactions with T cells, ECM, and cytokines in the lymph node tissues. The long-term goal is to generate single cell insights into B cell development in GCs of SIV+ and SIV- macaques in response to spontaneous activation and infections. The goal of this project is to define spatially resolved cellular interactions and cytokine/ECM gradients pixel-by-pixel in fixed macaque and human tissues. The hypothesis is that (1) spatial distributions of cell types, cytokines, and ECM of spontaneous GC activation are uniquely controlled by CD4/FDC and B cells in NHPs compared to humans, and (2) B-cell responses are spatiotemporally regulated by CD4/FDC cells in NHPs with and without SIV infections. The rationale for this hypothesis is based on the evidence that (1) IL- 10+ cells were spatially close to viral SIV-DNA+ lymphoid cells in SIV+ tissues and (2) heterogeneous GC activation and spatial GC organization maps in macaque and human tissues. The central hypothesis will be tested by pursuing two specific Aims. Aim 1 will evaluate the effect of multiplexed cytokine, ECM, and CD4/FDC interactions with B cell subsets in SIV- macaque (n=24 each) and HIV- human tissues (n=24). Aim 2 will evaluate cytokines, ECM, CD4/FDC, and B cell maturation in lymph node tissues of SIV- and SIV+ macaques (n=24). To accomplish these Aims, spatial cell phenotyping and cytokine gene expression profiling will be used to analyze B cell development and statistical comparisons of spatial cell neighboring features in macaque and human tissues. This project builds an interdisciplinary team integrating experts from spatial omics, NHP immunology and pathology, and bioinformatics. The proposed application is innovative because it uses cutting-edge technology to study spatial proteomics and transcriptomics of lymph node tissues of macaques and humans. This research is significant because it defines spatial GC organization to understand why SIV+ infections induce GC defects and how it deviates from human immune responses.

生发中心 (GC) 是次级淋巴器官的微观结构部位，但 GC 结构是 艾滋病患者由于失去支持性 CD4+ T 细胞而受损，导致免疫缺陷 HIV 感染者的抗逆转录病毒疗法的反应甚至无效。为了解决这个挑战， 恒河猴是成熟的非人灵长类动物模型 (NHP)，用于研究免疫发病机制 艾滋病病毒。然而，滤泡树突状细胞（FDC）、T细胞、B细胞、细胞外基质的空间协调 （ECM），以及猿猴免疫缺陷病毒（SIV）+感染的猕猴和SIV-的细胞因子调节 猕猴目前还不清楚。因此，迫切需要破译空间和时间 控制 GC 中单细胞水平的 (1) 细胞因子、(2) ECM 和 (3) T-B 细胞相互作用，以识别 (1) 自发激活和 (2) HIV/SIV 疾病免疫系统中 SIV 感染诱导的反应。 为了阐明 SIV+ 和 SIV- 猕猴以及人类捐赠者淋巴结组织的免疫调节，这项研究 该项目将利用空间蛋白质组学和转录分析来绘制 B 细胞亚群及其相互作用 淋巴结组织中的 T 细胞、ECM 和细胞因子。长期目标是产生单细胞 深入了解 SIV+ 和 SIV- 猕猴 GC 中 B 细胞响应自发激活和 感染。该项目的目标是定义空间分辨的细胞相互作用和细胞因子/ECM 在固定猕猴和人体组织中逐像素渐变。假设 (1) 的空间分布 自发 GC 激活的细胞类型、细胞因子和 ECM 由 CD4/FDC 和 B 细胞独特控制 与人类相比，NHP 中的 B 细胞反应受到 CD4/FDC 细胞的时空调节 感染或未感染 SIV 的 NHP。该假设的基本原理基于 (1) IL- 10+ 细胞在空间上接近 SIV+ 组织中的病毒 SIV-DNA+ 淋巴细胞，并且 (2) 异质 GC 猕猴和人体组织中的激活和空间 GC 组织图。中心假设将是 通过追求两个具体目标进行测试。目标 1 将评估多重细胞因子、ECM 和 CD4/FDC 与 SIV-猕猴（各 n=24）和 HIV-人类组织（n=24）中的 B 细胞亚群相互作用。目标2 将评估 SIV- 和 SIV+ 淋巴结组织中的细胞因子、ECM、CD4/FDC 和 B 细胞成熟度 猕猴 (n=24)。为了实现这些目标，空间细胞表型分析和细胞因子基因表达谱 将用于分析 B 细胞发育和空间细胞邻近特征的统计比较 猕猴和人体组织。该项目建立了一个跨学科团队，整合了来自空间领域的专家 组学、NHP 免疫学和病理学以及生物信息学。所提出的应用程序具有创新性，因为它 利用尖端技术研究淋巴结组织的空间蛋白质组学和转录组学 猕猴和人类。这项研究意义重大，因为它定义了空间 GC 组织来理解 为什么 SIV+ 感染会导致 GC 缺陷以及它如何偏离人类免疫反应。