Cellular and molecular architecture of granulomas in human tuberculous lymphadenitis

摘要 结核性淋巴结炎是最常见的肺外结核，影响约 突尼斯每年有2000人患有这种疾病，这对突尼斯的医疗保健来说是一种临床负担 系统。由于TBL患者合并肺结核的情况并不常见，细菌也很少。 通过显微镜检测，TBL代表了一种感染得到控制而尚未得到控制的结核病。 散布的，免疫功能受损的人除外。突尼斯的TBL通常是由 牛分枝杆菌或结核分枝杆菌，但不能从临床、放射学或 病态的。结核的组织学特征是肉芽肿，一种有组织的巨噬细胞集合， 淋巴细胞和其他类型的细胞，其功能是遏制和杀死入侵的病原体。在这里，我们将测试 假设阐明TBL肉芽肿的细胞和分子结构将导致 疾病生物标志物的识别和对T细胞反应调节机制的洞察 肉芽肿的抗微生物反应。我们建议研究细胞和分子的结构 单细胞RNA测序(scRNA-seq)与淋巴结空间序列相结合检测结直肠癌肉芽肿 作为常规临床护理的一部分而获得的标本。这个项目汇集了来自 突尼斯巴斯德研究所(IPT)的Benabdessalem实验室与来自加州大学洛杉矶分校的科学家合作，他们已经 合作生成了测量细胞类型和基因的空间分布的初步数据 肺结核病和TBL肉芽肿。在目标1中，我们将确定tbl的细胞和分子结构。 肉芽肿，识别与致病菌相关的抗微生物反应相关的生物标记物， 牛分枝杆菌与结核分枝杆菌的比较，以及对结核病的治疗效果的比较。在目标2中，我们将 通过抗菌药物检测T细胞在控制TBL肉芽肿感染中的作用 反应，包括与外周血中特定分枝杆菌抗原相关的反应。加在一起，这些 研究将提供新的洞察机制，通过T细胞反应和交叉点 肉芽肿组织影响宿主对牛分枝杆菌和结核分枝杆菌感染的控制。因此，我们对 TBL中的特定免疫生物特征将扩大我们对其免疫发病机制的理解，导致 改善了对TBL患者的护理。这一应用的一个重要部分是确保技术转让 以及从加州大学洛杉矶分校的莫德林实验室到IPT的Benabdessalem实验室的研究培训。

ABSTRACT Tuberculous lymphadenitis (TBL) is the most frequent form of extrapulmonary of TB affecting approximately 2000 individuals in Tunisia annually, such that the disease is a clinical burden on the Tunisian health care system. Since the occurrence of concomitant pulmonary TB in TBL patients is infrequent, and bacilli are rarely detected by microscopy, TBL represents a form of TB in which the infection is controlled and not yet disseminated, except in immunocompromised individuals. TBL in Tunisia is generally caused by Mycobacterium bovis or M. tuberculosis, yet cannot be distinguished clinically, radiographically, or pathologically. The histologic hallmark of TB is the granuloma, an organized collection of macrophages, lymphocytes, and other cell types that function to contain and kill invading pathogens. Here, we will test the hypothesis that elucidation of the cellular and molecular architecture of TBL granulomas will lead to the identification of disease biomarkers and insight into the mechanisms by which T cell responses regulates antimicrobial responses in granulomas. We propose to study the cellular and molecular architecture of granulomas in TBL by integrating single cell RNA-sequencing (scRNA-seq) with spatial-seq of lymph node specimens that are obtained as part of routine clinical care. This project brings together scientists from the Benabdessalem lab at the Institut Pasteur de Tunis (IPT) with scientists from UCLA, who have already collaborated to generate preliminary data measuring the spatial distribution of cell types and genes in pulmonary TB and TBL granulomas. In Aim 1, we will determine the cellular and molecular architecture of TBL granulomas, identifying biomarkers related to antimicrobial responses that correlate with the causative agent, M. bovis vs. M. tuberculosis, as well as comparing the response in TBL to pulmonary TB. In Aim 2, we will determine the contribution of T cells to control the infection in TBL granulomas through antimicrobial responses, including relevant correlates to specific mycobacterial antigens in peripheral blood. Together, these studies will provide new insight into the mechanisms by which the intersection of T cell responses and granuloma organization impacts host control of M. bovis vs. M. tuberculosis infection. As such, our studies of the specific immune biosignatures in TBL will expand our understanding of its immunopathogenesis, leading to improved care for patients with TBL. An important part of this application is to ensure transfer of technology and research training from the Modlin lab at UCLA to the Benabdessalem lab at IPT.