圣犹达儿童研究医院Paul G. Thomas课题组在研究中取得进展。他们的最新研究提出了TIRTL-seq：深度、定量和负担得起的配对TCR库测序。2025年11月24日出版的《自然—方法学》杂志发表了这项成果。

在这里，该团队提出了吞吐量密集型快速TCR库测序（TIRTL-seq），这是一种配对TCR库测序（TCR-seq）的实验和计算方法。TIRTL-seq基于384孔板中数百个TCR文库的并行生成，每个板的成本低于200美元，允许队列规模的配对TCR-seq研究。在纵向样本上，该研究组将TIRTL-seq与最先进的批量TCR-seq和10x Genomics Chromium技术进行比较，发现了感染后具有不同动态的严重急性呼吸综合征冠状病毒2和EB病毒特异性克隆扩增。TIRTL-seq提供了从单个细胞到每个样本数百万个T细胞的通用方案，同时提供精确的克隆频率估计和准确的TCR链配对，结合了批量和单细胞TCR-seq的优势。TIRTL-seq是一种在队列规模上对配对T细胞受体测序的高通量方法。

据介绍，T细胞的特异性由T细胞受体（TCR） α和β链序列决定。虽然批量TCR测序能够在没有链配对信息的情况下实现具有成本效益的曲目分析，但单细胞方法提供配对数据，但成本高且吞吐量有限。

附：英文原文

Title: TIRTL-seq: deep, quantitative and affordable paired TCR repertoire sequencing

Author: Pogorelyy, Mikhail V., Kirk, Allison M., Adhikari, Samir, Minervina, Anastasia A., Sundararaman, Balaji, Vegesana, Kasi, Brice, David C., Scott, Zachary B., Thomas, Paul G.

Issue&Volume: 2025-11-24

Abstract: The specificity of T cells is determined by T cell receptor (TCR) α and β chain sequences. While bulk TCR sequencing enables cost-effective repertoire profiling without chain pairing information, single-cell approaches provide paired data but are costly and limited in throughput. Here we present throughput-intensive rapid TCR library sequencing (TIRTL-seq), an experimental and computational methodology for paired TCR repertoire sequencing (TCR-seq). TIRTL-seq is based on the parallel generation of hundreds of TCR libraries in 384-well plates at less than US$200 per plate, allowing cohort-scale paired TCR-seq studies. We benchmarked TIRTL-seq against state-of-the-art bulk TCR-seq and 10x Genomics Chromium technologies on longitudinal samples and identified severe acute respiratory syndrome coronavirus 2- and Epstein–Barr virus-specific clonal expansions after infection with distinct dynamics. TIRTL-seq offers a universal protocol scalable from a single cell to millions of T cells per sample, simultaneously delivering both precise clonal frequency estimation and accurate TCR chain pairing, combining the strengths of bulk and single-cell TCR-seq. TIRTL-seq is a high-throughput method for paired T cell receptor sequencing at the cohort scale.

DOI: 10.1038/s41592-025-02907-9

Source: https://www.nature.com/articles/s41592-025-02907-9