Toulouse Institute for Infectious and Inflammatory Diseases (INFINITy), INSERM UMR1291, CNRS UMR5051, Toulouse III Paul Sabatier University, Toulouse, France; Ludwig Boltzmann Institute for Rare and Undiagnosed Diseases (LBI-RUD), Vienna, Austria.
CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria; Department of Structural and Computational Biology, Max Perutz Labs, University of Vienna, Vienna, Austria.
Cell Rep. 2021 Jul 6;36(1):109318. doi: 10.1016/j.celrep.2021.109318.
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.
免疫突触是一种复杂的结构，可将刺激信号解码为适应性的淋巴细胞反应。由于系统定量方法的发展，它是监测淋巴细胞活性的独特窗口。在这里，我们证明了高内涵成像在人类 T 细胞和自然杀伤 (NK) 细胞中的适用性，并开发了一种用于无偏分析高清晰度形态特征的流水线。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酶体颗粒的定位。对免疫缺陷个体的 CD8 T 细胞进行形态特征分析，可以区分 ARP2/3 亚基 ARPC1B 和 ARP2/3 激活蛋白威斯科特-奥尔德里奇综合征蛋白 (WASP) 在免疫突触组装中的作用。单细胞分析进一步确定了 ARPC1B 缺陷淋巴细胞中溶酶体颗粒和 F-actin 径向分布的解偶联。我们的研究为形态特征分析的发展提供了基础，可作为一种可扩展的方法来监测原代淋巴细胞的反应性，并确定淋巴细胞微结构的复杂方面。
