Department of Surgery, Cardinal Bernardin Cancer Center, Loyola University Chicago, Maywood, IL 60153, USA.
Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, USA; Harper Cancer Research Institute, University of Notre Dame, Notre Dame, IN 46556, USA.
Mol Ther. 2018 Apr 4;26(4):996-1007. doi: 10.1016/j.ymthe.2018.01.015. Epub 2018 Feb 2.
The use of T cell receptor (TCR) gene-modified T cells in adoptive cell transfer has had promising clinical success, but often, simple preclinical evaluation does not necessarily accurately predict treatment efficacy or safety. Preclinical studies generally evaluate one or a limited number of type 1 cytokines to assess antigen recognition. However, recent studies have implicated other "typed" T cells in effective anti-tumor/viral immunity, and limited functional evaluations may underestimate cross-reactivity. In this study, we use an altered peptide ligand (APL) model and multi-dimensional flow cytometry to evaluate polyfunctionality of TCR gene-modified T cells. Evaluating six cytokines and the lytic marker CD107a on a per cell basis revealed remarkably diverse polyfunctional phenotypes within a single T cell culture and among peripheral blood lymphocyte (PBL) donors. This polyfunctional assessment identified unexpected phenotypes, including cells producing both type 1 and type 2 cytokines, and highlighted interferon γ (IFNγ) antigen-reactive populations overlooked in our previous studies. Additionally, APLs skewed functional phenotypes to be less polyfunctional, which was not necessarily related to changes in TCR-peptide-major histocompatibility complex (pMHC) affinity. A better understanding of gene-modified T cell functional diversity may help identify optimal therapeutic phenotypes, predict clinical responses, anticipate off-target recognition, and improve the design and delivery of TCR gene-modified T cells.
T 细胞受体 (TCR) 基因修饰 T 细胞过继转移在临床应用中取得了可喜的成功，但通常情况下，简单的临床前评估并不一定能准确预测治疗效果或安全性。临床前研究通常评估一种或有限数量的 1 型细胞因子来评估抗原识别。然而，最近的研究表明其他“类型”的 T 细胞在有效的抗肿瘤/抗病毒免疫中起作用，有限的功能评估可能会低估交叉反应性。在这项研究中，我们使用改变的肽配体 (APL) 模型和多维流式细胞术来评估 TCR 基因修饰 T 细胞的多功能性。在单个 T 细胞培养物和外周血淋巴细胞 (PBL) 供体中，基于每个细胞评估六种细胞因子和裂解标志物 CD107a，发现了显著不同的多功能表型。这种多功能评估确定了意想不到的表型，包括产生 1 型和 2 型细胞因子的细胞，并突出了我们之前研究中忽略的干扰素 γ (IFNγ) 抗原反应性群体。此外，APL 使功能表型偏向于不那么多功能，这不一定与 TCR-肽-主要组织相容性复合物 (pMHC) 亲和力的变化有关。更好地了解基因修饰 T 细胞的功能多样性可能有助于确定最佳治疗表型，预测临床反应，预测非靶标识别，并改进 TCR 基因修饰 T 细胞的设计和递送。
