Morphological profiling of human T and NK lymphocytes by high-content cell imaging
Morphological profiling of human T and NK lymphocytes by high-content cell imaging
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DOI：
10.1016/j.celrep.2021.109318
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发表时间：
2021-07-06
期刊：
影响因子：
8.8
通讯作者：
Dupre, Loic
中科院分区：
文献类型：
--
作者：
German, Yolla;Vulliard, Loan;Dupre, Loic
免疫突触是一种复杂的结构，它将刺激信号解码为适应的淋巴细胞反应。由于系统定量方法的发展，这是监测淋巴细胞活性的唯一窗口。在这里，我们展示了高内容成像对人类T细胞和自然杀伤(NK)细胞的适用性，并开发了一种对高清晰度形态特征进行无偏见分析的管道。我们的方法揭示了肌动蛋白细胞骨架重塑的不同方面如何塑造免疫突触结构，并影响溶解颗粒的定位。免疫缺陷个体的CD8(+)T细胞的形态特征可以区分Arp2/3亚单位ARPC1B和Arp2/3激活因子Wiskott-Aldrich综合征蛋白(WASP)在免疫突触组装中的作用。单细胞分析进一步证实了ARPC1B缺陷淋巴细胞中溶解颗粒的解偶联和F-肌动蛋白的径向分布。我们的研究为发展形态图谱作为一种可扩展的方法来监测初级淋巴细胞的反应性和识别淋巴细胞微结构的复杂方面提供了基础。
The immunological synapse is a complex structure that decodes stimulatory signals into adapted lymphocyte responses. It is a unique window to monitor lymphocyte activity because of development of systematic quantitative approaches. Here we demonstrate the applicability of high-content imaging to human T and natural killer (NK) cells and develop a pipeline for unbiased analysis of high-definition morphological profiles. Our approach reveals how distinct facets of actin cytoskeleton remodeling shape immunological synapse architecture and affect lytic granule positioning. Morphological profiling of CD8(+) T cells from immunodeficient individuals allows discrimination of the roles of the ARP2/3 subunit ARPC1B and the ARP2/3 activator Wiskott-Aldrich syndrome protein (WASP) in immunological synapse assembly. Single-cell analysis further identifies uncoupling of lytic granules and F-actin radial distribution in ARPC1B-deficient lymphocytes. Our study provides a foundation for development of morphological profiling as a scalable approach to monitor primary lymphocyte responsiveness and to identify complex aspects of lymphocyte micro-architecture.