Establishment of a novel platform cell line for efficient and precise evaluation of T cell receptor functional avidity.
Establishment of a novel platform cell line for efficient and precise evaluation of T cell receptor functional avidity.
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DOI：
10.18632/oncotarget.26139
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发表时间：
2018-09-25
期刊：
影响因子：
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通讯作者：
Sugiyama H
中科院分区：
文献类型：
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作者：
Morimoto S;Fujiki F;Kondo K;Nakajima H;Kobayashi Y;Inatome M;Aoyama N;Nishida Y;Tsuboi A;Oka Y;Nishida S;Nakata J;Hosen N;Oji Y;Sugiyama H
使用T细胞受体（TCR）工程化的T细胞的连续性T细胞疗法是用于癌症治疗的有吸引力的策略，并且这种疗法的成功取决于转导的TCR对靶向肿瘤抗原的功能亲合力。因此，建立一种高效、准确的TCR功能亲合力评价方法一直是人们所期待的。在这里，我们展示了一种新的平台细胞系，命名为2D 3，它使转导的TCR的功能亲合力得到有效和精确的评估。在2D 3中，通过活化T细胞核因子（NFAT）经由TCR信号传导激活驱动的绿色荧光蛋白（GFP）报告基因的表达，可以容易地评估转导的TCR的精确TCR功能亲合力。将HLA-A*24：02限制性肾母细胞瘤基因1（WT 1）特异性CD 8+细胞毒性T淋巴细胞（CTL）的四种不同TCR转导到2D 3细胞中，并评价这四种TCR的功能亲合力。这些TCR的评价功能亲合力与TCR转导的CD 8 + T细胞响应WT 1抗原的细胞增殖、细胞因子产生和WT 1特异性细胞毒性正相关。这些结果表明，2D 3细胞系是一种新的和稳定的工具，可用于有效和精确地评估分离和转导的TCR的功能亲合力，在开发基于TCR的免疫疗法。
Adoptive T-cell therapy with T cell receptor (TCR) -engineered T cells is an attractive strategy for cancer treatment and the success in this therapy is dependent on the functional avidity of the transduced TCRs against targeted tumor antigens. Therefore, the establishment of the methodology of the efficient and precise evaluation of TCR functional avidity has been awaited. Here, we show a novel platform cell line, named 2D3, which enables the functional avidity of transduced TCRs to be evaluated efficiently and precisely. In the 2D3, the precise TCR functional avidity of transduced TCRs is easily evaluable by the expression of green fluorescent protein (GFP) reporter gene driven by nuclear factor of activated T cells (NFAT) activation via TCR signaling. Four different TCRs of HLA-A*24:02-restricted Wilms’ tumor gene 1 (WT1)-specific CD8+ cytotoxic T lymphocytes (CTLs) were transduced into 2D3 cells and the functional avidities of these four TCRs were evaluated. The evaluated functional avidity of these TCRs positively correlated with cell proliferation, cytokine production, and WT1-specific cytotoxicity of the TCR-transduced CD8+ T cells in response to WT1 antigen. These results showed that 2D3 cell line was a novel and stable tool useful for the efficient and precise evaluation of the functional avidity of isolated and transduced TCRs in developing TCR-based immunotherapy.