Interrogating monocyte-stromal cell crosstalk in the intestine

巨噬细胞是肠粘膜中含量最丰富的白细胞之一，在维持组织动态平衡和损伤、损伤或炎症后的修复方面发挥着重要作用。然而，它们也与炎症性肠病(IBD)的病理有关，在炎症性肠病中，它们似乎是疾病的驱动力。重要的是，我们已经证明促炎巨噬细胞和稳态/支持分解的巨噬细胞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，单核细胞的分化在进入肠道黏膜时立即发生，并受到微生物区系的影响，但控制单核细胞分化的确切因素仍然难以捉摸。粘膜的外渗和迁移无疑会涉及到与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，但这种串扰的性质及其如何影响单核细胞分化仍是完全未知的。此外，微生物群及其衍生物是如何影响这种串扰的，目前还不清楚。在这个项目中，学生将使用一种共培养系统来询问单核细胞和肠道基质细胞之间的串扰，使用小鼠和人类细胞，以及这种串扰在整个生命过程和随后的炎症挑战中可能会发生什么变化。此外，我们将确定这种情况在生物失调的背景下可能会发生什么变化。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图谱。学生将接受所有这些技术以及统计、演示技能、写作技能和数据管理方面的全面培训。

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.