Tumor-infiltrating lymphocytes (TILs) are a potent source for obtaining tumor-reactive T cell receptors (TCRs). Although comprehensive methods to analyze the TCR repertoire in TILs have been reported, the evaluation system for TCR-reactivity to endogenously expressed antigen in tumor cells remains laborious and time consuming. Consequently, very limited numbers of TCRs in TILs have been analyzed for their reactivity to tumor cells. In this study, we developed an efficient evaluation system for TCR function designated c-FIT (comprehensive functional investigation of TCRs) to analyze TCR reactivity. The c-FIT system enabled us to analyze up to 90 TCRs for their reactivity to tumor cells by a single assay within a month. Using c-FIT, we analyzed 70 TCRs of CD8⁺ TILs derived from two breast cancer patients and obtained 23 TCRs that reacted to tumor cells. Surprisingly, although two TCRs were HLA class I-restricted, the remaining 21 TCRs were non-HLA-restricted. Thus, c-FIT can be applied for monitoring multiple conventional and unconventional antigen-specific killer T cells in TILs, leading to the development of new designs for more effective T-cell-based immunotherapies.

中文翻译：

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使用新系统的 TCR 功能分析揭示了人乳腺癌浸润淋巴细胞中的多个非常规肿瘤反应性 T 细胞

肿瘤浸润淋巴细胞 (TIL) 是获得肿瘤反应性 T 细胞受体 (TCR) 的有效来源。尽管已经报道了分析 TIL 中 TCR 库的综合方法，但 TCR 对肿瘤细胞中内源性表达抗原的反应性的评估系统仍然费时费力。因此，已经分析了 TIL 中数量非常有限的 TCR 对肿瘤细胞的反应性。在这项研究中，我们开发了TCR功能指定C-FIT一个有效的评估系统（ç omprehensive ˚F unctional我的nvestigation ŧCRs) 来分析 TCR 反应性。c-FIT 系统使我们能够在一个月内通过一次检测分析多达 90 个 TCR 对肿瘤细胞的反应性。使用 c-FIT，我们分析了来自两名乳腺癌患者的70 个 CD8 ⁺ TILs 的 TCR，并获得了 23 个与肿瘤细胞反应的 TCR。令人惊讶的是，尽管两个 TCR 受 HLA I 类限制，但其余 21 个 TCR 不受 HLA 限制。因此，c-FIT 可用于监测 TIL 中的多种常规和非常规抗原特异性杀伤 T 细胞，从而为更有效的基于 T 细胞的免疫疗法开发新设计。