Tracing spatial organization of germinal centers in rhesus macaques

生发中心(GC)是次级淋巴器官的微结构部位，而GC结构是 艾滋病患者由于失去支持的CD4+T细胞而受损，导致免疫功能低下 艾滋病毒感染者在抗逆转录病毒治疗中的反应，甚至无效。为了解决这一挑战， 恒河猴是一种成熟的非人灵长类动物模型，用于研究糖尿病的免疫发病机制。 爱滋病毒。然而，卵泡树突状细胞(FDCs)、T细胞、B细胞、细胞外基质的空间配位 (ECM)，以及SIV+感染猴和SIV感染猕猴的细胞因子调节。 对猕猴的了解还不是很清楚。因此，迫切需要破译空间和时间 控制(1)细胞因子，(2)细胞外基质，和(3)T-B细胞在单个细胞水平上的相互作用，以识别GC (1)在HIV/SIV病的免疫系统中自发激活和(2)SIV感染诱导的反应。 为了阐明SIV+和SIV-猕猴和人类捐赠者淋巴组织的免疫调节， 该项目将利用空间蛋白质组学和转录图谱来绘制B细胞亚群及其相互作用 淋巴组织中有T细胞、细胞外基质和细胞因子。长期目标是产生单个细胞 对SIV+和SIV-猕猴GC中B细胞发育的洞察 感染。这个项目的目标是定义空间分辨的细胞相互作用和细胞因子/细胞外基质 固定猕猴和人体组织中逐个像素的渐变。假设是(1)空间分布 自发性GC激活的细胞类型、细胞因子和ECM由CD4/FDC和B细胞唯一控制 与人类相比，以及(2)B细胞反应在时空上受CD4/FDC细胞的调节 有和没有SIV感染的NHP。这一假说的理论基础是基于以下证据：(1)IL- SIV+组织中10+细胞与病毒SIV-DNA+淋巴样细胞空间位置接近；(2)异质性GC 猕猴和人类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来检验。目标1将评估多重细胞因子、ECM和 在SIV猕猴(n=24)和HIV-人类组织(n=24)中，CD4/FDC与B细胞亚群相互作用。目标2 将评估SIV-和SIV+淋巴组织中的细胞因子、ECM、CD4/FDC和B细胞成熟情况 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 将用于分析B细胞的发育和空间单元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，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。拟议的应用程序具有创新性，因为它 利用尖端技术研究小鼠淋巴组织的空间蛋白质组学和转录组学 猕猴和人类。这项研究具有重要意义，因为它定义了要理解的空间GC组织 为什么SIV+感染会导致GC缺陷，以及它如何偏离人类的免疫反应。

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.