Malte Lehmann, Benjamin Weixler, Sefer Elezkurtaj, Christopher Loddenkemper, Anja A Kühl, Britta Siegmund
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引用次数: 0
Abstract
Background and aims: Fistula formation is a major complication in Crohn's disease [CD] and the role of the immune cell compartment remains to be elucidated. Thus, we compared the immune cell compartment of CD fistula to inflammatory CD colitis using imaging mass cytometry [IMC] and immunofluorescence.
Methods: A 36-marker panel including structural, functional, and lineage markers for use in IMC was established. This panel was applied to analyse paraffin-embedded CD fistula tract [n = 11], CD colitis [n = 10], and colon samples from non-inflamed controls [n = 12]. Computational methods for cell segmentation, dimensionality reduction, and cell type clustering were used to define cell populations for cell frequency, marker distribution, and spatial neighbourhood analysis. Multiplex immunofluorescence was used for higher resolution spatial analysis.
Results: Analysis of cell frequencies in CD fistulas compared to CD colitis and control colonic samples revealed a significant increase in neutrophils, effector cytotoxic T cells, and inflammatory macrophages in CD fistula samples, whereas regulatory T cells were decreased. Neutrophils in CD fistula expressed significantly more matrix metalloproteinase 9 [MMP9], correlating with extracellular matrix remodelling. Neighbourhood analysis revealed a strong association between MMP9+ neutrophils and effector cytotoxic T cells in both CD fistulas and colitis.
Conclusions: This study presents the first highly multiplexed single cell analysis of the immune cell compartment of CD fistulas and their spatial context. It links immune cell dynamics, particularly MMP9+ neutrophils, to extracellular matrix remodelling in CD fistulas, offering insights into the complex network of cellular interactions and potential therapeutic targets for CD complications.
背景和目的:瘘管形成是克罗恩病(CD)的主要并发症,而免疫细胞区系的作用仍有待阐明。因此,我们使用成像质控细胞仪和免疫荧光技术比较了 CD 性瘘管和 CD 性结肠炎的免疫细胞区系:方法:我们建立了一个包括结构、功能和系谱标记在内的 36 个标记面板,用于成像质谱(IMC)。该面板用于分析石蜡包埋的CD瘘管(11人)、CD结肠炎(10人)和非炎症对照组的结肠样本(12人)。使用细胞分割、降维和细胞类型聚类的计算方法来定义细胞群,以进行细胞频率、标记物分布和空间邻域分析。多重免疫荧光用于更高分辨率的空间分析:结果:CD瘘与CD结肠炎和对照结肠样本的细胞频率分析表明,CD瘘样本中的中性粒细胞、效应细胞毒性T细胞和炎性巨噬细胞显著增加,而调节性T细胞减少。CD瘘管中的中性粒细胞表达的基质金属蛋白酶9(MMP9)明显增多,这与细胞外基质重塑有关。邻近分析显示,在CD瘘和结肠炎中,MMP9+中性粒细胞与效应细胞毒性T细胞之间存在密切联系:本研究首次对 CD 性瘘管的免疫细胞区系及其空间环境进行了高度复用的单细胞分析。它将免疫细胞动态(尤其是 MMP9+ 中性粒细胞)与 CD 性瘘管中细胞外基质的重塑联系起来,深入揭示了细胞相互作用的复杂网络以及 CD 并发症的潜在治疗靶点。