Division of Gastroenterology, The Institute for Adult Diseases, Asahi Life Foundation, Tokyo, Japan.
Department of Gastroenterology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
J Crohns Colitis. 2018 Nov 9;12(10):1219-1231. doi: 10.1093/ecco-jcc/jjy088.
Disturbance of intestinal homeostasis is associated with the development of inflammatory bowel disease [IBD], and TGF-β signalling impairment in mononuclear phagocytes [MPs] causes murine colitis with goblet cell depletion. Here, we examined an organoid-MP co-culture system to study the role of MPs in intestinal epithelial differentiation and homeostasis.
Intestinal organoids were co-cultured with lamina propria leukocytes and bone marrow-derived dendritic cells [BMDCs] from CD11c-cre Tgfbr2fl/fl mice. Organoid-MP adhesive interactions were evaluated by microscopy, RT-PCR, and flow cytometry. Murine colitis models (dextran sodium sulphate [DSS], CD11c-cre Tgfbr2fl/fl, T-cell-transfer) were used for histological and immunohistochemical analysis. Anti-E-cadherin antibody treatment or CD11c+-cell-specific CDH1 gene deletion were performed for E-cadherin neutralization or knockout. Colonic biopsies from patients with ulcerative colitis were analysed by flow cytometry.
Intestinal organoids co-cultured with CD11c+ lamina propria leukocytes or BMDCs from CD11c-cre Tgfbr2fl/fl mice showed morphological changes and goblet cell depletion with Notch signal activation, analogous to CD11c-cre Tgfbr2fl/fl colitis. E-cadherin was upregulated in CD11c+ MPs, especially CX3CR1+CCR2+ monocytes, of CD11c-cre Tgfbr2fl/fl mice. E-cadherin-mediated BMDC adhesion promoted Notch activation and cystic changes in organoids. Anti-E-cadherin antibody treatment attenuated colitis in CD11c-cre Tgfbr2fl/fl and T-cell-transferred mice. In addition, E-cadherin deletion in CD11c+ cells attenuated colitis in both CD11c-cre Tgfbr2fl/fl and DSS-treated mice. In patients with ulcerative colitis, E-cadherin expressed by intestinal CD11c+ leukocytes was enhanced compared with that in healthy controls.
E-cadherin-mediated MP-epithelium adhesion is associated with the development of colitis, and blocking these adhesions may have therapeutic potential for IBD.
肠道内稳态的紊乱与炎症性肠病[IBD]的发生有关，单核吞噬细胞[MPs]中的 TGF-β信号转导受损会导致小鼠结肠炎伴杯状细胞耗竭。在这里，我们研究了器官样-MP 共培养系统，以研究 MPs 在肠道上皮细胞分化和内稳态中的作用。
将肠类器官与来自 CD11c-cre Tgfbr2fl/fl 小鼠的固有层白细胞和骨髓来源的树突状细胞[BMDC]共培养。通过显微镜、RT-PCR 和流式细胞术评估类器官-MP 黏附相互作用。使用葡聚糖硫酸钠[DSS]、CD11c-cre Tgfbr2fl/fl、T 细胞转移的小鼠结肠炎模型进行组织学和免疫组织化学分析。进行抗 E-钙黏蛋白抗体处理或 CD11c+-细胞特异性 CDH1 基因敲除，以实现 E-钙黏蛋白中和或敲除。通过流式细胞术分析溃疡性结肠炎患者的结肠活检。
与 CD11c+固有层白细胞或来自 CD11c-cre Tgfbr2fl/fl 小鼠的 BMDC 共培养的肠类器官显示出形态学变化和杯状细胞耗竭，同时 Notch 信号激活，类似于 CD11c-cre Tgfbr2fl/fl 结肠炎。E-钙黏蛋白在 CD11c+MPs，特别是 CD11c-cre Tgfbr2fl/fl 小鼠中的 CX3CR1+CCR2+单核细胞中上调。E-钙黏蛋白介导的 BMDC 黏附促进了 Notch 的激活和类器官的囊性改变。抗 E-钙黏蛋白抗体处理可减轻 CD11c-cre Tgfbr2fl/fl 和 T 细胞转移小鼠的结肠炎。此外，CD11c+细胞中的 E-钙黏蛋白缺失可减轻 CD11c-cre Tgfbr2fl/fl 和 DSS 处理小鼠的结肠炎。在溃疡性结肠炎患者中，与健康对照相比，肠 CD11c+白细胞表达的 E-钙黏蛋白增强。
E-钙黏蛋白介导的 MP-上皮黏附与结肠炎的发生有关，阻断这些黏附可能为 IBD 提供治疗潜力。
