Profiling lymphocyte interactions at the single-cell level by microfluidic cell pairing
Profiling lymphocyte interactions at the single-cell level by microfluidic cell pairing
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DOI：
10.1038/ncomms6940
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发表时间：
2015-01-01
影响因子：
16.6
通讯作者：
Voldman, Joel
中科院分区：
文献类型：
--
作者：
Dura, Burak;Dougan, Stephanie K.;Voldman, Joel
建立成功的免疫反应需要细胞间的相互作用，其中抗原提呈的性质决定了功能结果。然而，研究这些相互作用的方法受到吞吐量有限和缺乏对细胞配对的控制的影响。在这里，我们描述了一种微流控平台，它实现了具有定义的接触时间的淋巴细胞的高通量确定配对，从而允许在受控微环境中准确地评估每对淋巴细胞的早期激活事件。更重要的是，该平台允许同时捕获来自双方的动态过程和静态参数，从而能够在单个实验中对数百对淋巴细胞相互作用进行成对相关的多参数分析。利用我们的平台，我们研究了CD8 T细胞(OT-1和TRP1跨核(TN))的早期激活动力学，并研究了T细胞激活的异质性程度和多个读数的相关性。这些结果使我们的平台成为定量研究淋巴细胞相互作用的一个很有前途的工具。
Establishing a successful immune response requires cell-cell interactions, where the nature of antigen presentation dictates functional outcomes. Methods to study these interactions, however, suffer from limited throughput and a lack of control over cell pairing. Here we describe a microfluidic platform that achieves high-throughput deterministic pairing of lymphocytes with a defined contact time, thereby allowing accurate assessment of early activation events for each pair in controlled microenvironments. More importantly, the platform allows the capture of dynamic processes and static parameters from both partners simultaneously, thus enabling pairwise-correlated multiparametric profiling of lymphocyte interactions over hundreds of pairs in a single experiment. Using our platform, we characterized early activation dynamics of CD8 Tcells (OT-1 and TRP1 transnuclear (TN)) and investigated the extent of heterogeneity in T-cell activation and the correlation of multiple readouts. The results establish our platform as a promising tool for quantitative investigation of lymphocyte interactions.