Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS) coined by inflammation and neurodegeneration. The actual cause of the neurodegenerative component of the disease is however unclear. We investigated here the direct and differential effects of inflammatory mediators on human neurons. We used embryonic stem cell-derived (H9) human neuronal stem cells (hNSC) to generate neuronal cultures. Neurons were subsequently treated with tumour necrosis factor alpha (TNFα), interferon gamma (IFNγ), granulocyte–macrophage colony-stimulating factor (GM-CSF), interleukin 17A (IL-17A) and interleukin 10 (IL-10) separately or in combination. Immunofluorescence staining and quantitative polymerase chain reaction (qPCR) were used to assess cytokine receptor expression, cell integrity and transcriptomic changes upon treatment. H9-hNSC-derived neurons expressed cytokine receptors for IFNγ, TNFα, IL-10 and IL-17A. Neuronal exposure to these cytokines resulted in differential effects on neurite integrity parameters with a clear decrease for TNFα- and GM-CSF-treated neurons. The combinatorial treatment with IL-17A/IFNγ or IL-17A/TNFα induced a more pronounced effect on neurite integrity. Furthermore, combinatorial treatments with two cytokines induced several key signalling pathways, i.e. NFκB-, hedgehog and oxidative stress signalling, stronger than any of the cytokines alone. This work supports the idea of immune-neuronal crosstalk and the need to focus on the potential role of inflammatory cytokines on neuronal cytoarchitecture and function.

Graphical Abstract

中文翻译：

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与多发性硬化症相关的炎症细胞因子直接诱导人类神经元神经元细胞结构的改变

多发性硬化症 (MS) 是一种由炎症和神经变性引起的中枢神经系统 (CNS) 慢性炎症性疾病。然而，该疾病的神经退行性成分的真正原因尚不清楚。我们在这里研究了炎症介质对人类神经元的直接和差异性影响。我们使用胚胎干细胞衍生的 (H9) 人神经元干细胞 (hNSC) 来生成神经元培养物。随后分别用肿瘤坏死因子 α (TNFα)、干扰素 γ (IFNγ)、粒细胞-巨噬细胞集落刺激因子 (GM-CSF)、白细胞介素 17A (IL-17A) 和白细胞介素 10 (IL-10) 单独或单独治疗神经元。组合。使用免疫荧光染色和定量聚合酶链反应（qPCR）来评估治疗后细胞因子受体表达、细胞完整性和转录组变化。H9-hNSC 衍生的神经元表达 IFNγ、TNFα、IL-10 和 IL-17A 的细胞因子受体。神经元暴露于这些细胞因子会对神经突完整性参数产生不同的影响，其中 TNFα 和 GM-CSF 处理的神经元的神经突完整性参数明显降低。IL-17A/IFNγ或IL-17A/TNFα的组合治疗对神经突完整性产生更显着的影响。此外，两种细胞因子的组合治疗诱导了几个关键信号传导途径，即 NFκB-、hedgehog 和氧化应激信号传导，比任何单独的细胞因子更强。这项工作支持免疫神经元串扰的想法，以及需要关注炎症细胞因子对神经元细胞结构和功能的潜在作用。

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