B-Cell Commitment to IL-10 Production: The VertX Il10(egfp) Mouse.
B-Cell Commitment to IL-10 Production: The VertX Il10(egfp) Mouse.
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DOI：
10.1007/978-1-0716-1237-8_19
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发表时间：
2021
期刊：
影响因子：
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通讯作者：
Sartor RB
中科院分区：
文献类型：
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作者：
Oka A;Liu B;Herzog JW;Sartor RB
尽管炎性细胞因子IL-10在调节B细胞功能中起关键作用，但通过细胞内IL-10染色检测产生IL-10的B细胞需要多个步骤和繁琐的准备。相比之下，2009年开发的Il10-EGFP报告小鼠模型(VertX)可以更容易、更快地检测到产生IL-10的B细胞，并有可能在不需要膜通透性和体外激活的情况下分离活细胞。尽管检测IL-10+细胞更简单，但有几个细微差别很重要。例如，含有甲醇的缓冲液会删除GFP信号，而长期固定可以保持GFP信号的强度，但会减少其他细胞内信号(FOXP3等)。在此，我们提供了肠道B细胞GFP检测的优化和改进方法，以及VertX小鼠固有层、脾、肠系膜淋巴结、腹膜和血细胞的分离技术。
Although the inflammatory cytokine IL-10 is pivotal in regulatory B cell function, detecting IL-10-producing B cells by intracellular IL-10 staining requires multiple steps and tedious preparation. In contrast, the Il10-eGFP-reporter mouse model (VertX), generated in 2009, allows easier and quicker detection of IL-10-producing B cells with the possibility of sorting viable cells without membrane permeabilization and ex vivo activation. Even though detecting IL-10+ cells is simpler, several nuances are important. For example, methanol-containing buffers delete GFP signal while long-term fixation can maintain GFP intensity but decreases other intracellular signals (FOXP3 etc.). Here, we provide optimized and improved protocols for GFP detection in intestinal B cells and isolation techniques of lamina propria, spleen, mesenteric lymph node, peritoneum and blood cells from VertX mice.