Chatterjee Ayan, Bandyopadhyay Aniket, Maiti Tapas Kumar, Kanti Bhattacharyya Tarun
Advanced Technology Development Centre, Indian Institute of Technology Kharagpur, Kharagpur, India.
Institute of Cell Dynamics and Imaging, University of Münster, Münster, Germany.
Microsyst Nanoeng. 2024 Nov 26;10(1):178. doi: 10.1038/s41378-024-00769-3.
Cellular communication at the single-cell level holds immense potential for uncovering response heterogeneity in immune cell behaviors. However, because of significant size diversity among different immune cell types, controlling the pairing of cells with substantial size differences remains a formidable challenge. We developed a microfluidic platform for size-selective pairing (SSP) to pair single cells with up to a fivefold difference in size, achieving over 40% pairing efficiency. We used SSP to investigate the real-time effects of combinatorial immunotherapeutic stimulation on macrophage T-cell interactions at the single-cell level via fluorescence microscopy and microfluidic sampling. While combinatorial activation involving toll-like receptor (TLR) agonists and rapamycin (an mTOR inhibitor) has improved therapeutic efficacy in mice, its clinical success has been limited. Here, we investigated immune synaptic interactions and outcomes at the single-cell level in real time and compared them with bulk-level measurements. Our findings, after tracking and computationally analyzing the effects of sequential and spatiotemporal stimulations of primary mouse macrophages, suggest a regulatory role of rapamycin in dampening inflammatory outputs in T cells.
单细胞水平上的细胞通讯在揭示免疫细胞行为中的反应异质性方面具有巨大潜力。然而，由于不同免疫细胞类型之间存在显著的大小差异，控制大小差异较大的细胞配对仍然是一项艰巨的挑战。我们开发了一种用于大小选择性配对（SSP）的微流控平台，以配对大小差异高达五倍的单细胞，实现了超过40%的配对效率。我们使用SSP通过荧光显微镜和微流控采样在单细胞水平上研究组合免疫治疗刺激对巨噬细胞与T细胞相互作用的实时影响。虽然涉及 toll 样受体（TLR）激动剂和雷帕霉素（一种 mTOR 抑制剂）的组合激活在小鼠中提高了治疗效果，但其临床成功有限。在这里，我们实时研究了单细胞水平上的免疫突触相互作用和结果，并将其与整体水平的测量结果进行了比较。我们在追踪并通过计算分析原代小鼠巨噬细胞的顺序和时空刺激的影响后发现，雷帕霉素在抑制T细胞炎症输出方面具有调节作用。
