GigaMune, Inc., South San Francisco, CA, USA.
Division of Cancer Sciences, University of Manchester, Manchester, UK.
Nat Biotechnol. 2020 May;38(5):609-619. doi: 10.1038/s41587-020-0438-y. Epub 2020 Mar 16.
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.
经基因工程改造表达抗原特异性 T 细胞受体 (TCR) 的 T 细胞是治疗病毒感染和癌症的有效疗法。然而，由于 T 细胞受体库的大小和复杂性以及原代 T 细胞的操作挑战，高效鉴定临床候选 TCR 变得复杂。在这里，我们提出了一种从多样化的人类 T 细胞受体库中鉴定具有高功能亲和力的 TCR 的高通量方法。该方法使用大规模平行微流控技术，从数百万个原代 T 细胞中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 文库可用于使用肽主要组织相容性复合物结合和细胞激活进行抗原反应性 TCR 的重复筛选和淘选。我们从六位健康供体中捕获了超过 290 万个天然配对的 TCRαβ 克隆型，并鉴定了罕见的 (<0.001% 频率) 病毒抗原反应性 TCR。我们还从黑色素瘤患者的肿瘤浸润淋巴细胞样本中进行挖掘，并鉴定了几种肿瘤特异性 TCR，这些 TCR 在原代 T 细胞中表达后可导致肿瘤细胞杀伤。
