High-dimensional analysis of intestinal immune cells during helminth infection
High-dimensional analysis of intestinal immune cells during helminth infection
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DOI：
10.7554/elife.51678
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发表时间：
2020-02-11
期刊：
影响因子：
7.7
通讯作者：
Mayer, Johannes U.
中科院分区：
文献类型：
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作者：
Ferrer-Font, Laura;Mehta, Palak;Mayer, Johannes U.
由于强烈的抗 2 型寄生虫免疫反应会驱动粘液产生、组织重塑和免疫细胞浸润，因此从感染蠕虫的小鼠肠道中分离单细胞非常困难。通过对标准肠道消化方案的系统优化，我们能够成功地从严重感染的十二指肠中分离出数百万个免疫细胞。为了验证这些细胞能够准确地表达肠道免疫反应，我们使用高维光谱流式细胞仪面板对它们进行了分析，并通过共聚焦显微镜证实了我们的发现。我们的细胞分离方案和高维分析使我们能够识别出蠕虫感染整个过程中抗寄生虫免疫反应的许多已知标志，并有可能加速以前无法实现的局部蠕虫免疫反应的单细胞发现。
Single cell isolation from helminth-infected murine intestines has been notoriously difficult, due to the strong anti-parasite type 2 immune responses that drive mucus production, tissue remodeling and immune cell infiltration. Through the systematic optimization of a standard intestinal digestion protocol, we were able to successfully isolate millions of immune cells from the heavily infected duodenum. To validate that these cells gave an accurate representation of intestinal immune responses, we analyzed them using a high-dimensional spectral flow cytometry panel and confirmed our findings by confocal microscopy. Our cell isolation protocol and high-dimensional analysis allowed us to identify many known hallmarks of anti-parasite immune responses throughout the entire course of helminth infection and has the potential to accelerate single-cell discoveries of local helminth immune responses that have previously been unfeasible.