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MATE-Seq:微流控抗原-TCR 衔接测序。

MATE-Seq: microfluidic antigen-TCR engagement sequencing.

机构信息

Division of Chemistry and Chemical Engineering, California Institute of Technology, 1200 East California Blvd, Pasadena, CA 91125, USA.

出版信息

Lab Chip. 2019 Sep 10;19(18):3011-3021. doi: 10.1039/c9lc00538b.

Abstract

Adaptive immunity is based on peptide antigen recognition. Our ability to harness the immune system for therapeutic gain relies on the discovery of the T cell receptor (TCR) genes that selectively target antigens from infections, mutated proteins, and foreign agents. Here we present a method that selectively labels peptide antigen-specific CD8+ T cells using magnetic nanoparticles functionalized with peptide-MHC tetramers, isolates these specific cells within an integrated microfluidic device, and directly amplifies the TCR genes for sequencing. Critically, the identity of the peptide recognized by the TCR is preserved, providing the link between peptide and gene. The platform requires inputs on the order of just 100 000 CD8+ T cells, can be multiplexed for simultaneous analysis of multiple peptides, and performs sorting and isolation on chip. We demonstrate 1000-fold sensitivity enhancement of detecting antigen-specific TCRs relative to bulk analysis and simultaneous capture of two virus antigen-specific TCRs from a population of T cells.

摘要

适应性免疫基于肽抗原识别。我们利用免疫系统获得治疗效果的能力依赖于发现 T 细胞受体 (TCR) 基因,这些基因可以选择性地针对感染、突变蛋白和外来物质的抗原。在这里,我们提出了一种使用肽-MHC 四聚体功能化的磁性纳米颗粒选择性标记肽抗原特异性 CD8+ T 细胞的方法,在集成微流控装置中分离这些特异性细胞,并直接扩增 TCR 基因进行测序。关键是,TCR 识别的肽的身份得到保留,为肽与基因之间提供了联系。该平台所需的输入量仅为 100000 个 CD8+ T 细胞左右,可以进行多重分析,同时分析多个肽,并在芯片上进行分选和分离。我们证明,与批量分析相比,检测抗原特异性 TCR 的灵敏度提高了 1000 倍,并且可以从 T 细胞群体中同时捕获两种病毒抗原特异性 TCR。

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