T cells engineered to express antigen-specific T cell receptors (TCRs) are potent therapies for viral infections and cancer. However, efficient identification of clinical candidate TCRs is complicated by the size and complexity of T cell repertoires and the challenges of working with primary T cells. Here we present a high-throughput method to identify TCRs with high functional avidity from diverse human T cell repertoires. The approach used massively parallel microfluidics to generate libraries of natively paired, full-length TCRαβ clones, from millions of primary T cells, which were then expressed in Jurkat cells. The TCRαβ-Jurkat libraries enabled repeated screening and panning for antigen-reactive TCRs using peptide major histocompatibility complex binding and cellular activation. We captured more than 2.9 million natively paired TCRαβ clonotypes from six healthy human donors and identified rare (<0.001% frequency) viral-antigen-reactive TCRs. We also mined a tumor-infiltrating lymphocyte sample from a patient with melanoma and identified several tumor-specific TCRs, which, after expression in primary T cells, led to tumor cell killing.

中文翻译：

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天然配对人类 TCRαβ 曲目的大规模并行审讯和挖掘。

经工程改造以表达抗原特异性 T 细胞受体 (TCR) 的 T 细胞是病毒感染和癌症的有效疗法。然而，临床候选 TCR 的有效鉴定因 T 细胞库的大小和复杂性以及使用原代 T 细胞的挑战而变得复杂。在这里，我们提出了一种高通量方法，用于从不同的人类 T 细胞库中识别具有高功能亲和力的 TCR。该方法使用大规模并行微流体从数百万个原代 T 细胞生成天然配对的全长 TCRαβ 克隆文库，然后在 Jurkat 细胞中表达。TCRαβ-Jurkat 文库能够使用肽主要组织相容性复合物结合和细胞活化重复筛选和淘选抗原反应性 TCR。我们捕获了2个以上。来自 6 名健康人类供体的 900 万个天然配对 TCRαβ 克隆型，并鉴定出罕见的（<0.001% 频率）病毒抗原反应性 TCR。我们还从一名黑色素瘤患者身上提取了一个肿瘤浸润淋巴细胞样本，并鉴定了几种肿瘤特异性 TCR，这些 TCR 在原代 T 细胞中表达后会导致肿瘤细胞死亡。