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）是次级淋巴器官的显微结构部位，但GC结构是 艾滋病患者由于失去支持性CD 4 + T细胞而受损，导致免疫缺陷， HIV感染者的抗逆转录病毒治疗反应甚至无效。为了解决这一挑战， 恒河猴（rhesus macaques）是研究免疫发病机制的成熟的非人灵长类动物模型（NHP）。 艾滋病。然而，滤泡树突状细胞（FDCs）、T细胞、B细胞、细胞外基质 (ECM)猴免疫缺陷病毒（SIV）+感染的猕猴和SIV-感染的猕猴的细胞因子调节 对猕猴的了解还不清楚。因此，迫切需要破译空间和时间 在GC中在单细胞水平上控制（1）细胞因子、（2）ECM和（3）T-B细胞相互作用，以鉴定 (1)自发激活和（2）SIV感染诱导的HIV/SIV疾病免疫系统应答。 为了阐明SIV+和SIV-猕猴和人供体淋巴结组织中的免疫调节， 一个项目将利用空间蛋白质组学和转录谱来绘制B细胞亚群及其相互作用 与淋巴结组织中的T细胞、ECM和细胞因子。长期目标是产生单细胞 对SIV+和SIV-猕猴GC中响应自发激活的B细胞发育的见解， 感染.本项目的目标是确定空间分辨的细胞相互作用和细胞因子/ECM 在固定的猕猴和人体组织中逐像素的梯度。假设是：（1）空间分布 自发性GC活化的细胞类型、细胞因子和ECM由CD 4/FDC和B细胞唯一控制 与人类相比，NHP中的B细胞应答，以及（2）在NHP中，B细胞应答受CD 4/FDC细胞的时空调节。 有和没有SIV感染的NHP。这一假设的基本原理是基于以下证据：（1）IL-10是一种细胞因子， 10+细胞在空间上接近SIV+组织中的病毒SIV-DNA+淋巴样细胞， 激活和空间GC组织图。核心假设是 通过追求两个具体目标进行测试。目的1将评估复合细胞因子、ECM和 在SIV-猕猴（各n=24）和HIV-人组织（n = 24）中，CD 4/FDC与B细胞亚群的相互作用。目的2 将评估SIV-和SIV+淋巴结组织中的细胞因子、ECM、CD 4/FDC和B细胞成熟 猕猴（n=24）。为了实现这些目标，空间细胞表型和细胞因子基因表达谱分析 将用于分析B细胞发育和空间细胞相邻特征的统计比较， 猕猴和人体组织。该项目建立了一个跨学科的团队， 组学、NHP免疫学和病理学以及生物信息学。该应用程序是创新的，因为它 使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是有意义的，因为它定义了空间GC组织， 为什么SIV+感染诱导GC缺陷以及它如何偏离人类免疫反应。