Interrogating monocyte-stromal cell crosstalk in the intestine

巨噬细胞是肠粘膜中最丰富的白细胞之一，在维持组织稳态和损伤、损伤或炎症后的修复中起着重要作用。然而，它们也与炎症性肠病（IBD）的病理有关，在那里它们似乎是疾病的驱动因素。重要的是，我们已经证明了促炎巨噬细胞和稳态/促溶解巨噬细胞都来源于相同的经典单核细胞前体。虽然我们的工作已经确定单核细胞在进入肠道黏膜时立即发生分化，并受到微生物群存在的影响，但控制单核细胞分化的确切因素仍然难以捉摸。粘膜的外溢和迁移无疑涉及不同结构的非造血细胞（包括内皮细胞、周细胞、成纤维细胞和上皮细胞）的串扰，但这种串扰的性质以及它如何影响单核细胞分化还完全不清楚。此外，微生物群及其衍生物如何影响这种串音仍然知之甚少。在这个项目中，学生将使用一个共培养系统来研究单核细胞和肠间质细胞之间的串扰，使用小鼠和人类细胞，以及这种串扰在整个生命过程和炎症挑战后如何变化。此外，我们将确定这在生态失调的情况下如何改变。技术将包括多参数流式细胞术、共聚焦显微镜、细胞因子/趋化因子阵列和转录谱分析。学生将在所有这些技术以及统计学、演讲技巧、写作技巧和数据管理方面接受全面的培训。

Macrophages are one of the most abundant leucocytes in the intestinal mucosa where they play fundamental roles in maintaining tissue homeostasis and repair following injury, insult or inflammation. However, they are also implicated in the pathology of inflammatory bowel disease (IBD), where they appear to drive disease. Importantly, we have shown that both pro-inflammatory macrophages and homeostatic/pro-resolution macrophages derive from the same classical monocyte precursor. While our work has established that the differentiation of monocytes occurs immediately as they enter the gut mucosa and is influenced by the presence of the microbiota, the exact factors that control monocyte differentiation remain elusive. Extravasation and migration in the mucosa will undoubtedly involve crosstalk with different structural, non-haematopoietic cells, including endothelia, pericytes, fibroblasts and epithelial cells, yet the nature of this crosstalk and how it influences monocyte differentiation is completely unexplored. Moreover, how the microbiota and its derivatives influence this crosstalk remains poorly understood.In this project, the student will use a co-culture system to interrogate the crosstalk between monocytes and intestinal stromal cells, using both mouse and human cells, and how this might change across the life course and following inflammatory challenge. Moreover, we will determine how this might change in the context of dysbiosis. Techniques will include multi-parameter flow cytometry, confocal microscopy, cytokine/chemokine arrays and transcriptional profiling. The student will be fully trained in all these techniques as well as in statistics, presentation skills, writing skills, and data management.