École Normale Supérieure, UMR 8640, Laboratoire PASTEUR, Département de Chimie, PSL Research University, Sorbonne Université, CNRS, 75005 Paris, France.
Institut Curie, U932, Immunology and Cancer, INSERM, 75005 Paris, France.
Cell Rep Methods. 2022 Nov 9;2(11):100335. doi: 10.1016/j.crmeth.2022.100335. eCollection 2022 Nov 21.
The immune synapse is the tight contact zone between a lymphocyte and a cell presenting its cognate antigen. This structure serves as a signaling platform and entails a polarization of intracellular components necessary to the immunological function of the cell. While the surface properties of the presenting cell are known to control the formation of the synapse, their impact on polarization has not yet been studied. Using functional lipid droplets as tunable artificial presenting cells combined with a microfluidic pairing device, we simultaneously observe synchronized synapses and dynamically quantify polarization patterns of individual B cells. By assessing how ligand concentration, surface fluidity, and substrate rigidity impact lysosome polarization, we show that its onset and kinetics depend on the local antigen concentration at the synapse and on substrate rigidity. Our experimental system enables a fine phenotyping of monoclonal cell populations based on their synaptic readout.
免疫突触是淋巴细胞与呈递其同源抗原的细胞之间的紧密接触区域。该结构作为信号平台，需要细胞的免疫功能所必需的细胞内成分的极化。虽然已经知道呈递细胞的表面特性控制着突触的形成，但它们对极化的影响尚未得到研究。我们使用功能性脂滴作为可调谐的人工呈递细胞，并结合微流配对装置，同时观察到同步的突触，并动态定量单个 B 细胞的极化模式。通过评估配体浓度、表面流动性和基质刚性如何影响溶酶体极化，我们表明其起始和动力学取决于突触处的局部抗原浓度和基质刚性。我们的实验系统能够基于突触的读出对单克隆细胞群体进行精细表型分析。
