Flow cytometric gating for spleen monocyte and DC subsets: differences in autoimmune NOD mice and with acute inflammation.
Flow cytometric gating for spleen monocyte and DC subsets: differences in autoimmune NOD mice and with acute inflammation.
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DOI：
10.1016/j.jim.2015.08.015
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发表时间：
2016-05
影响因子：
2.2
通讯作者：
Tarbell KV
中科院分区：
文献类型：
--
作者：
Dong MB;Rahman MJ;Tarbell KV
由于多色流式细胞术、APC 群体基因表达分析以及小鼠与人类 APC 群体功能相关性的进步，现在人们更好地了解了抗原呈递细胞 (APC) 在自身免疫和其他炎症性疾病发病机制中的作用。传统和浆细胞样树突细胞亚群（cDC1、cDC2、pDC）和单核细胞衍生群体的简单但信息丰富的命名法融合了这些进展，但准确的亚群识别至关重要。模糊的门控方案和炎症条件下细胞表面标记物的改变可能会使研究之间的结果进行比较变得困难。急性炎症（如 TLR 配体刺激）和慢性炎症（如在小鼠自身免疫模型中发现的）都可以改变 DC 子集门控。在这里，我们使用体内 CpG 刺激作为急性炎症的例子，并使用非肥胖糖尿病 (NOD) 小鼠作为慢性炎症的模型来解决这些问题。我们提供了流式细胞术抗体组和门控方案，可在稳态和 CpG 刺激后区分脾脏中的 2 个单核细胞和 3 个 DC 亚群。使用这种方法，我们观察到先前报道的 NOD DC 组成的差异，以及新发现的 NOD 单核细胞衍生的 DC 数量的增加。最后，我们建立了 DC phosphflow 协议来测量细胞内蛋白质的磷酸化状态，并用它来确认已识别子集的功能差异。因此，我们提出了用于区分有或没有与 NOD 小鼠相关的炎症和/或自身免疫的单核细胞和 DC 群体的优化方法。
The role of antigen presenting cells (APCs) in the pathogenesis of autoimmune and other inflammatory diseases is now better understood due to advances in multicolor flow cytometry, gene expression analysis of APC populations, and functional correlation of mouse to human APC populations. A simple but informative nomenclature of conventional and plasmacytoid dendritic cell subsets (cDC1, cDC2, pDC) and monocyte-derived populations incorporates these advances, but accurate subset identification is critical. Ambiguous gating schemes and alterations of cell surface markers in inflammatory condition can make comparing results between studies difficult. Both acute inflammation, such as TLR–ligand stimulation, and chronic inflammation as found in mouse models of autoimmunity can alter DC subset gating. Here, we address these issues using in vivo CpG stimulation as an example of acute inflammation and the non-obese diabetic (NOD) mouse as a model of chronic inflammation. We provide a flow cytometric antibody panel and gating scheme that differentiate 2 monocytic and 3 DC subsets in the spleen both at steady state and after CpG stimulation. Using this method, we observed differences in the composition of NOD DCs that have been previously reported, and newly identified increases in the number of NOD monocyte-derived DCs. Finally, we established a protocol for DC phosphoflow to measure the phosphorylation state of intracellular proteins, and use it to confirm functional differences in the identified subsets. Therefore, we present optimized methods for distinguishing monocytic and DC populations with and without inflammation and/or autoimmunity associated with NOD mice.
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影响因子：
32.4
作者：
Serbina, NV;Salazar-Mather, TP;Pamer, EG
通讯作者：
Pamer, EG
影响因子：
82.9
作者：
Diana, Julien;Simoni, Yannick;Lehuen, Agnes
通讯作者：
Lehuen, Agnes
影响因子：
3.7
作者：
Larsen J;Weile C;Antvorskov JC;Engkilde K;Nielsen SM;Josefsen K;Buschard K
通讯作者：
Buschard K
DOI：
10.1084/jem.20041930
发表时间：
2005-04-04
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Asselin-Paturel C;Brizard G;Chemin K;Boonstra A;O'Garra A;Vicari A;Trinchieri G
通讯作者：
Trinchieri G
影响因子：
7.3
作者：
Price JD;Tarbell KV
通讯作者：
Tarbell KV