Functional dynamics of TB granuloma architecture

项目总结/摘要 了解和消除结核病取决于了解肉芽肿中的宿主和病原体动力学， 其中M.结核病的感染情况。尽管付出了巨大的努力， 组成，空间相互作用和机制，决定结果，如病理 肉芽肿坏死知之甚少。对肉芽肿了解有限的一种解释是， 大多数研究是在单独的实验系统中进行的。我们成立了一个财团 在三个系统中进行和整合结核肉芽肿的研究：人类、恒河猴和新的 小鼠品系。在这三个系统中，我们将解决结核肉芽肿生物学中的几个主要问题： 结核肉芽肿细胞组成的多样性程度如何？什么规范了细胞贩运， 结核肉芽肿的空间相互作用I型干扰素（TIFN）和干扰素的作用是什么？ γ-干扰素（IFNg）在调节和决定细胞运输、分化、空间关系和活化中的作用 结核病肉芽肿的症状什么是坏死的病原体和宿主决定因素，一种病理结果， 肺结核肉芽肿我们研究这些和其他问题的主要模式将是高参数 这三个物种的结核肉芽肿的多重免疫染色，因为这将提供必要的了解， 免疫细胞亚群之间的空间关系。多重免疫染色将补充 恒河猴肉芽肿细胞动力学的实时成像研究和策略性定时转移 在恒河猴和不同品系的小鼠中的标记细胞。多重免疫染色的价值 研究将通过在单细胞水平上对解离细胞群的深入表征来进一步加强。 水平，使用高参数流式细胞术和单细胞转录组学。一个独特的重要贡献 我们提出的项目的一个重要部分是计算整合来自人类，恒河猴和新的 小鼠品系。通过整合多物种数据，我们将确定结核肉芽肿的特征， 这三个物种都有共同之处，我们将确定每个物种肉芽肿所特有的因素， 指导进一步修改，以提高动物模型研究的实用性。整合来自 三个物种将允许恒河猴的研究与人类样本进行比较， 新的小鼠品系;这将提高对人类病理学研究的理解和解释， 提高研究的价值，可以利用小鼠的独特优势， 问题研究总之，我们的研究将促进结核病生物学的知识和理解，并指导 制定干预措施，以改善治疗结果，限制肺损伤，并改善肺组织 肺结核患者体内的修复。

Project Summary/Abstract Understanding and eliminating TB depends on understanding the host and pathogen dynamics in granulomas, where the outcomes of M. tuberculosis infection are determined. Despite substantial efforts, the cellular composition, spatial interactions, and mechanisms that determine outcomes such as pathologic granuloma necrosis are poorly understood. One explanation for the limited understanding of granulomas is that most studies are performed in a single experimental system in isolation. We have formed a consortium to perform and integrate studies of TB granulomas in three systems: humans, rhesus macaques, and new strains of mice. In these three systems, we will address several major questions in TB granuloma biology: what is the extent of diversity in cell composition in TB granulomas? What regulates cell trafficking and spatial interactions in TB granulomas? What are the roles of type I interferons (TIIFN) and interferon gamma (IFNg) in regulating and determining cell trafficking, differentiation, spatial relationships, and activation states in TB granulomas? What are the pathogen and host determinants of necrosis, a pathologic outcome, in TB granulomas? Our primary mode for investigating these and other questions will be high-parameter multiplex immunostaining of TB granulomas from the three species, as this will provide essential insight into the spatial relationships between immune cell subsets. Multiplex immunostaining will be complemented by studies of live imaging of cell dynamics in rhesus macaque granulomas and by strategically-timed transfers of labeled cells in rhesus macaques and diverse strains of mice. The value of the multiplex immunostaining studies will be further enhanced by deep characterization of dissociated cell populations at the single-cell level, using high parameter flow cytometry and single-cell transcriptomics. A uniquely important contribution of our proposed project is the computational integration of data from humans, rhesus macaques, and new strains of mice. By integrating multispecies data, we will identify the features of TB granulomas that are common to all three species, and we will identify factors that are unique to granulomas in each species, to guide further modifications to improve the utility of studies in animal models. Integration of the data from the three species will allow studies in rhesus macaques to be compared with those of human samples and new strains of mice; this will improve the understanding and interpretation of human pathology studies, and enhance the value of studies that can take advantage of using the unique advantages of mice for mechanistic studies. Together, our studies will advance the knowledge and understanding of TB biology, and guide development of interventions to improve treatment outcomes, limit lung damage, and improve lung tissue repair in people suffering from TB.