Bioluminescence and Near-infrared Imaging of Optic Neuritis and Brain Inflammation in the EAE Model of Multiple Sclerosis in Miceh
Bioluminescence and Near-infrared Imaging of Optic Neuritis and Brain Inflammation in the EAE Model of Multiple Sclerosis in Miceh
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DOI：
10.3791/55321
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发表时间：
2017-03-01
影响因子：
1.2
通讯作者：
Tegeder, Irmgard
中科院分区：
文献类型：
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作者：
Schmitz, Katja;Tegeder, Irmgard
SJL/J小鼠的实验性自身免疫性脑脊髓炎（EAE）是复发-缓解型多发性硬化症（RRMS）的模型。描述运动功能缺陷的临床EAE评分是脊髓免疫介导炎症的基本读数。然而，评分和体重不允许在体内评估脑炎症和视神经炎。后者在大约2/3的MS患者中是早期和频繁的表现。在这里，我们展示了生物发光和近红外实时成像的方法，利用活体成像系统评估EAE诱发的视神经炎、脑炎症和血脑屏障（BBB）破坏。由氧化酶激活的生物发光底物主要表现为视神经炎。该信号是特异性的，并允许可视化药物效果和疾病时间进程，这与临床评分相一致。聚乙二醇荧光纳米颗粒在血管内停留较长时间，用于评估血脑屏障的完整性。近红外成像显示在疾病高峰期血脑屏障泄漏。眼睛周围的信号最强。近红外基质金属蛋白酶底物被用来评估eae诱发的炎症。自动荧光干扰信号，需要光谱解混才能定量。总体而言，生物发光成像是评估eae相关视神经炎和药物效果的可靠方法，在信号特异性、稳健性、易于量化和成本方面优于近红外技术。
Experimental autoimmune encephalomyelitis (EAE) in SJL/J mice is a model for relapsing-remitting multiple sclerosis (RRMS). Clinical EAE scores describing motor function deficits are basic readouts of the immune-mediated inflammation of the spinal cord. However, scores and body weight do not allow for an in vivo assessment of brain inflammation and optic neuritis. The latter is an early and frequent manifestation in about 2/3 of MS patients. Here, we show methods for bioluminescence and near-infrared live imaging to assess EAE evoked optic neuritis, brain inflammation, and blood-brain barrier (BBB) disruption in living mice using an in vivo imaging system. A bioluminescent substrate activated by oxidases primarily showed optic neuritis. The signal was specific and allowed the visualization of medication effects and disease time courses, which paralleled the clinical scores. Pegylated fluorescent nanoparticles that remained within the vasculature for extended periods of time were used to assess the BBB integrity. Near-infrared imaging revealed a BBB leak at the peak of the disease. The signal was the strongest around the eyes. A near-infrared substrate for matrix metalloproteinases was used to assess EAE-evoked inflammation. Auto-fluorescence interfered with the signal, requiring spectral unmixing for quantification. Overall, bioluminescence imaging was a reliable method to assess EAE-associated optic neuritis and medication effects and was superior to the near-infrared techniques in terms of signal specificity, robustness, ease of quantification, and cost.