Ella Lemelbaum Institute for Immuno-Oncology, Sheba Medical Center, Ramat Gan, Israel.
Laboratory of Immunology, MIGAL-Galilee Research Institute, Kiryat Shmona, Israel.
Front Immunol. 2021 Jan 7;11:584148. doi: 10.3389/fimmu.2020.584148. eCollection 2020.
Adoptive T cell therapy (ACT) holds great promise for cancer treatment. One approach, which has regained wide interest in recent years, employs antitumor T cells isolated from tumor lesions ("tumor-infiltrating lymphocytes" or TIL). It is now appreciated that a considerable proportion of anti-melanoma TIL recognize new HLA-binding peptides resulting from somatic mutations, which occurred during tumor progression. The clinical efficacy of TIL can potentially be improved their genetic modification, designed to enhance their survival, homing capacity, resistance to suppression, tumor killing ability and additional properties of clinical relevance. Successful implementation of such gene-based strategies critically depends on efficient and reproducible protocols for gene delivery into clinical TIL preparations. Here we describe an optimized protocol for the retroviral transduction of TIL. As the experimental system we employed anti-melanoma TIL cultures prepared from four patients, recombinant retrovirus encoding an anti-CD19 chimeric antigen receptor (CAR) as a model gene of interest and CD19+ and CD19- human cell lines serving as target cells. Transduction on day 7 of the rapid expansion protocol (REP) resulted in 69 ± 8% CAR positive TIL. Transduced, but not untransduced TIL, from the four patients responded robustly to CD19+, but not CD19- cell lines, as judged by substantial secretion of IFN-γ following co-culture. In light of the rekindled interest in antitumor TIL, this protocol can be incorporated into a broad range of gene-based approaches for improving the in-vivo survival and functionality of TIL in the clinical setting.
过继性 T 细胞疗法（ACT）在癌症治疗中具有巨大的潜力。近年来，一种方法重新引起了广泛的关注，该方法采用从肿瘤病灶中分离出的抗肿瘤 T 细胞（“肿瘤浸润淋巴细胞”或 TIL）。现在人们认识到，相当一部分抗黑色素瘤 TIL 识别新的 HLA 结合肽，这些肽是在肿瘤进展过程中由体细胞突变产生的。TIL 的临床疗效可以通过其遗传修饰来提高，目的是增强其生存能力、归巢能力、抗抑制能力、杀伤肿瘤能力和其他具有临床相关性的特性。成功实施此类基于基因的策略取决于将基因有效且可重复地递送到临床 TIL 制剂中的方案。在这里，我们描述了一种优化的 TIL 逆转录病毒转导方案。我们采用了从四位患者制备的抗黑色素瘤 TIL 培养物作为实验系统，用编码抗 CD19 嵌合抗原受体（CAR）的重组逆转录病毒作为感兴趣的模型基因，用 CD19+和 CD19-人细胞系作为靶细胞。在快速扩增方案（REP）的第 7 天进行转导，导致 69±8%的 CAR 阳性 TIL。从四位患者中转导的，但未转导的 TIL，在与共培养后，IFN-γ大量分泌的情况下，对 CD19+细胞系而非 CD19-细胞系表现出强烈的反应。鉴于对抗肿瘤 TIL 的重新关注，该方案可被纳入一系列广泛的基于基因的方法，以提高 TIL 在临床环境中的体内存活和功能。
