Design of T-cell receptor libraries with diverse binding properties to examine adoptive T-cell responses
Design of T-cell receptor libraries with diverse binding properties to examine adoptive T-cell responses
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DOI：
10.1038/gt.2012.80
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发表时间：
2013-06-01
期刊：
影响因子：
5.1
通讯作者：
Kranz, D. M.
中科院分区：
文献类型：
--
作者：
Chervin, A. S.;Stone, J. D.;Kranz, D. M.
连续性T细胞疗法在治疗癌症和病毒性疾病方面显示出显著的前景。将抗原特异性T细胞受体（TCR）引入离体活化的T细胞的一种方法被设计为克服阻止内源性T细胞库的应答的中枢耐受机制。研究表明，使用针对I类主要组织相容性复合物抗原的较高亲和力的TCR可以驱动CD4（+）和CD8（+）T细胞的活性，但是控制体内活性的最佳TCR结合的规则是未知的。在这里，我们描述了一个高通量的“反向生物化学”平台，其中具有广泛的结合特性的TCR库相同的抗原被引入到T细胞和过继转移到小鼠与抗原阳性肿瘤。从肿瘤浸润淋巴细胞（TIL）或淋巴器官提取RNA允许高通量测序以确定在体内选择哪些TCR。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缺失。相反，这些相同的高亲和力TCR变体优先在肿瘤中的CD4（+）T细胞内表达，表明它们在抗原特异性肿瘤控制中起作用。因此，这些发现揭示了转导的TCR的亲和力控制了转移的T细胞的存活和肿瘤浸润。因此，TCR文库策略使得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。
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.