Multimodal analysis of ocular inflammation using the endotoxin-induced uveitis mouse model.
Multimodal analysis of ocular inflammation using the endotoxin-induced uveitis mouse model.
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DOI：
10.1242/dmm.022475
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发表时间：
2016-04
影响因子：
4.3
通讯作者：
Dick AD
中科院分区：
文献类型：
--
作者：
Chu CJ;Gardner PJ;Copland DA;Liyanage SE;Gonzalez-Cordero A;Kleine Holthaus SM;Luhmann UF;Smith AJ;Ali RR;Dick AD
啮齿动物内毒素诱导的葡萄膜炎 (EIU) 是急性 Toll 样受体 4 (TLR4) 介导的器官炎症的模型，已用于模拟人类前葡萄膜炎、检查白细胞运输和测试新型抗炎疗法。由于需要对每只眼睛的组织学切片进行手动、非特异性细胞计数评分作为疾病严重程度的衡量标准，因此更广泛的采用受到限制。在这里，我们描述了一种使用眼解剖和多模式组织分析的全面有效的技术。这样可以通过炎症浸润的多色流式细胞术分析、眼部上清液的蛋白质分析以及同一只眼睛的残余组织的 qPCR 来进行匹配的疾病评分。可以识别细胞群的动态变化，并将其映射到模型过程中趋化因子和细胞因子的变化。为了验证该技术，在 Ccl2 或 Ccr2 缺陷的小鼠中，重组白细胞介素 10 对白细胞浸润的剂量反应性抑制以及单核细胞 (CD11b+Ly6C+) 浸润的选择性抑制得到了证实。该模型首次使用光学相干断层扫描 (OCT) 来对浸润玻璃体细胞进行体内成像，并与 CD11b+Ly6G+ 计数相关联，以提供眼组织中细胞群的另一种独特测量方法。 EIU 的多模态组织分析被提出作为一个新标准，以改进和拓宽该模型的应用。摘要：针对内毒素诱导的葡萄膜炎的精细平台方法，使用多模式离体和体内评估，以改善这种分隔组织炎症眼模型的定量和表型分析。
Endotoxin-induced uveitis (EIU) in rodents is a model of acute Toll-like receptor 4 (TLR4)-mediated organ inflammation, and has been used to model human anterior uveitis, examine leukocyte trafficking and test novel anti-inflammatory therapeutics. Wider adoption has been limited by the requirement for manual, non-specific, cell-count scoring of histological sections from each eye as a measure of disease severity. Here, we describe a comprehensive and efficient technique that uses ocular dissection and multimodal tissue analysis. This allows matched disease scoring by multicolour flow cytometric analysis of the inflammatory infiltrate, protein analysis on ocular supernatants and qPCR on remnant tissues of the same eye. Dynamic changes in cell populations could be identified and mapped to chemokine and cytokine changes over the course of the model. To validate the technique, dose-responsive suppression of leukocyte infiltration by recombinant interleukin-10 was demonstrated, as well as selective suppression of the monocyte (CD11b+Ly6C+) infiltrate, in mice deficient for either Ccl2 or Ccr2. Optical coherence tomography (OCT) was used for the first time in this model to allow in vivo imaging of infiltrating vitreous cells, and correlated with CD11b+Ly6G+ counts to provide another unique measure of cell populations in the ocular tissue. Multimodal tissue analysis of EIU is proposed as a new standard to improve and broaden the application of this model. Summary: A refined platform approach for endotoxin-induced uveitis, using multimodal ex vivo and in vivo assessment, to improve quantification and phenotyping in this ocular model of compartmentalised tissue inflammation.