Department of Biochemistry, University of Illinois, Urbana, IL 61801, USA.
Gene Ther. 2013 Jun;20(6):634-44. doi: 10.1038/gt.2012.80. Epub 2012 Oct 11.
Adoptive T-cell therapies have shown significant promise in the treatment of cancer and viral diseases. One approach, which introduces antigen-specific T-cell receptors (TCRs) into ex vivo activated T cells, is designed to overcome central tolerance mechanisms that prevent responses by endogenous T-cell repertoires. Studies have suggested that use of higher-affinity TCRs against class I major histocompatibility complex antigens could drive the activity of both CD4(+) and CD8(+) T cells, but the rules that govern the TCR binding optimal for in vivo activity are unknown. Here, we describe a high-throughput platform of 'reverse biochemistry' whereby a library of TCRs with a wide range of binding properties to the same antigen is introduced into T cells and adoptively transferred into mice with antigen-positive tumors. Extraction of RNA from tumor-infiltrating lymphocytes (TILs) or lymphoid organs allowed high-throughput sequencing to determine which TCRs were selected in vivo. The results showed that CD8(+) T cells expressing the highest-affinity TCR variants were deleted in both the TIL population and in peripheral lymphoid tissues. In contrast, these same high-affinity TCR variants were preferentially expressed within CD4(+) T cells in the tumor, suggesting they had a role in antigen-specific tumor control. The findings thus revealed that the affinity of the transduced TCRs controlled the survival and tumor infiltration of the transferred T cells. Accordingly, the TCR library strategy enables rapid assessment of TCR-binding properties that promote peripheral T-cell survival and tumor elimination.
过继性 T 细胞疗法在癌症和病毒性疾病的治疗中显示出巨大的应用潜力。其中一种方法是将抗原特异性 T 细胞受体（TCR）引入体外激活的 T 细胞中，旨在克服阻止内源性 T 细胞库产生反应的中枢耐受机制。研究表明，使用针对 I 类主要组织相容性复合物抗原的高亲和力 TCR 可以驱动 CD4(+)和 CD8(+)T 细胞的活性，但尚不清楚调控 TCR 结合以实现体内活性的规则。在这里，我们描述了一种高通量的“反向生物化学”平台，该平台使用具有广泛结合同一抗原的 TCR 文库，并将其导入 T 细胞中，然后将其过继转移到具有抗原阳性肿瘤的小鼠体内。从肿瘤浸润淋巴细胞（TIL）或淋巴器官中提取 RNA ，允许进行高通量测序，以确定哪种 TCR 在体内被选择。结果表明，表达最高亲和力 TCR 变体的 CD8(+)T 细胞在 TIL 群体和外周淋巴组织中均被删除。相比之下，这些相同的高亲和力 TCR 变体在肿瘤内的 CD4(+)T 细胞中优先表达，表明它们在抗原特异性肿瘤控制中发挥作用。因此，研究结果表明，转导的 TCR 亲和力控制着转移 T 细胞的存活和肿瘤浸润。因此，TCR 文库策略能够快速评估促进外周 T 细胞存活和肿瘤消除的 TCR 结合特性。
