Division of Genetics, Department of Medicine, Howard Hughes Medical Institute, Brigham and Women's Hospital, Boston, MA 02115, USA; Department of Genetics, Harvard University Medical School, Boston, MA, USA.
Division of Genetics, Department of Medicine, Howard Hughes Medical Institute, Brigham and Women's Hospital, Boston, MA 02115, USA; Department of Genetics, Harvard University Medical School, Boston, MA, USA; Department of Pathology, Massachusetts General Hospital, Boston, MA, USA.
Cell. 2019 Aug 8;178(4):1016-1028.e13. doi: 10.1016/j.cell.2019.07.009.
T cell recognition of specific antigens mediates protection from pathogens and controls neoplasias, but can also cause autoimmunity. Our knowledge of T cell antigens and their implications for human health is limited by the technical limitations of T cell profiling technologies. Here, we present T-Scan, a high-throughput platform for identification of antigens productively recognized by T cells. T-Scan uses lentiviral delivery of antigen libraries into cells for endogenous processing and presentation on major histocompatibility complex (MHC) molecules. Target cells functionally recognized by T cells are isolated using a reporter for granzyme B activity, and the antigens mediating recognition are identified by next-generation sequencing. We show T-Scan correctly identifies cognate antigens of T cell receptors (TCRs) from viral and human genome-wide libraries. We apply T-Scan to discover new viral antigens, perform high-resolution mapping of TCR specificity, and characterize the reactivity of a tumor-derived TCR. T-Scan is a powerful approach for studying T cell responses.
T 细胞识别特定抗原可介导对病原体的保护并控制肿瘤，但也可能导致自身免疫。我们对 T 细胞抗原及其对人类健康的影响的了解受到 T 细胞分析技术的技术限制。在这里，我们展示了 T-Scan，这是一种用于鉴定 T 细胞可有效识别的抗原的高通量平台。T-Scan 使用慢病毒将抗原文库递送至细胞内，以进行内源性加工和主要组织相容性复合体 (MHC) 分子的呈递。通过用于颗粒酶 B 活性的报告基因来分离被 T 细胞功能识别的靶细胞，并通过下一代测序鉴定介导识别的抗原。我们证明 T-Scan 可以正确识别来自病毒和人类全基因组文库的 T 细胞受体 (TCR) 的同源抗原。我们应用 T-Scan 来发现新的病毒抗原，进行 TCR 特异性的高分辨率作图，并表征肿瘤衍生 TCR 的反应性。T-Scan 是研究 T 细胞反应的强大方法。
