Identification of Rare High-Avidity, Tumor-Reactive CD8+ T Cells by Monomeric TCR-Ligand Off-Rates Measurements on Living Cells
Identification of Rare High-Avidity, Tumor-Reactive CD8+ T Cells by Monomeric TCR-Ligand Off-Rates Measurements on Living Cells
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DOI：
10.1158/0008-5472.can-14-3516
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发表时间：
2015-05-15
期刊：
影响因子：
11.2
通讯作者：
Rufer, Nathalie
中科院分区：
文献类型：
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作者：
Hebeisen, Michael;Schmidt, Julien;Rufer, Nathalie
t细胞受体（TCR）对多肽- mhc （pMHC）呈递的抗原肽的亲和力是细胞介导免疫的关键参数。然而，大多数肿瘤抗原特异性CD8(+) T细胞的一个基本特征是这种亲和力较低。在这项研究中，我们解决了识别和选择肿瘤特异性CD8(+) T细胞的需求，这是癌症患者过继细胞治疗的最大兴趣。为了鉴定这些罕见的细胞，我们开发了一种多肽- mhc多聚体技术，该技术使用可逆的Ni2+-硝基三乙酸组氨酸标签（NTAmers）。NTAmers是高度稳定的，但加入咪唑后，它们迅速衰变为pMHC单体，允许基于流式细胞术的测量活CD8(+) T细胞的单体TCR-pMHC解离率在宽光谱上。我们记录了NTAmer动力学结果与表面等离子体共振得到的结果之间的强相关性。使用缺乏CD8与pMHC结合的NTAmers，我们发现CD8本身稳定了TCR-pMHC复合物，将解离半衰期延长了几倍。值得注意的是，我们的NTAmer技术准确地预测了从癌症患者中分离出来的大量肿瘤特异性T细胞的功能。总的来说，我们的研究结果表明，NTAmers是从患者身上分离罕见的高亲和力细胞毒性T细胞的有效工具，可用于癌症治疗的过继性治疗。(c) 2015年aacr。
The avidity of the T-cell receptor (TCR) for antigenic peptides presented by the peptide-MHC (pMHC) on cells is a key parameter for cell-mediated immunity. Yet a fundamental feature of most tumor antigen-specific CD8(+) T cells is that this avidity is low. In this study, we addressed the need to identify and select tumor-specific CD8(+) T cells of highest avidity, which are of the greatest interest for adoptive cell therapy in patients with cancer. To identify these rare cells, we developed a peptide-MHC multimer technology, which uses reversible Ni2+-nitrilotriacetic acid histidine tags (NTAmers). NTAmers are highly stable but upon imidazole addition, they decay rapidly to pMHC monomers, allowing flow-cytometric-based measurements of monomeric TCR-pMHC dissociation rates of living CD8(+) T cells on a wide avidity spectrum. We documented strong correlations between NTAmer kinetic results and those obtained by surface plasmon resonance. Using NTAmers that were deficient for CD8 binding to pMHC, we found that CD8 itself stabilized the TCR-pMHC complex, prolonging the dissociation half-life several fold. Notably, our NTAmer technology accurately predicted the function of large panels of tumor-specific T cells that were isolated prospectively from patients with cancer. Overall, our results demonstrated that NTAmers are effective tools to isolate rare high-avidity cytotoxic T cells from patients for use in adoptive therapies for cancer treatment. (C) 2015 AACR.