Chimeric antigen receptor T-cell (CAR-T) therapy has demonstrated potent clinical efficacy in the treatment of hematopoietic malignancies. However, the application of CAR-T in solid tumors has been limited due in part to the expression of inhibitory molecules in the tumor microenvironment, leading to T-cell exhaustion. To overcome this limitation, we have developed a synthetic T-cell receptor (TCR) that targets programmed death-ligand 1 (PD-L1), a molecule that is widely expressed in various solid tumors and plays a pivotal role in T-cell exhaustion. Our novel TCR platform is based on antibody-based binding domain, which is typically a single-chain variable fragment (scFv), fused to the γδ TCRs (TCRγδ). We have utilized the T-cell receptor alpha constant (TRAC) locus editing approach to express cell surface scFv of anti-PD-L1, which is fused to the constant region of the TCRγ or TCRδ chain in activated T cells derived from peripheral blood mononuclear cells (PBMCs). Our results indicate that these reconfigured receptors, both γ-TCRγδ and δ-TCRγδ, have the capability to transduce signals, produce inflammatory cytokines, degranulate and exert tumor killing activity upon engagement with PD-L1 antigen in vitro. Additionally, we have also shown that γ-TCRγδ exerted superior efficacy than δ-TCRγδ in in vivo xenograft model.

嵌合抗原受体T细胞（CAR-T）疗法在治疗造血系统恶性肿瘤方面已显示出强大的临床疗效。然而，CAR-T在实体瘤中的应用受到限制，部分原因是肿瘤微环境中抑制分子的表达，导致T细胞耗竭。为了克服这一限制，我们开发了一种针对程序性死亡配体 1 (PD-L1) 的合成 T 细胞受体 (TCR)，PD-L1 是一种在各种实体瘤中广泛表达的分子，在 T 细胞耗竭中发挥着关键作用。我们的新型 TCR 平台基于抗体结合域，通常是与 γδ TCR (TCRγδ) 融合的单链可变片段 (scFv)。我们利用 T 细胞受体 α 常数 (TRAC) 位点编辑方法来表达抗 PD-L1 的细胞表面 scFv，它与源自外周血单核细胞 (PBMC) 的活化 T 细胞中 TCRγ 或 TCRδ 链的恒定区融合。我们的结果表明，这些重新配置的受体（γ-TCRγδ 和 δ-TCRγδ）在体外与 PD-L1 抗原结合后具有转导信号、产生炎性细胞因子、脱颗粒和发挥肿瘤杀伤活性的能力。此外，我们还表明，在体内异种移植模型中，γ-TCRγδ 比 δ-TCRγδ 表现出更优异的功效。在体外与 PD-L1 抗原结合后脱颗粒并发挥肿瘤杀伤活性。此外，我们还表明，在体内异种移植模型中，γ-TCRγδ 比 δ-TCRγδ 表现出更优异的功效。在体外与 PD-L1 抗原结合后脱颗粒并发挥肿瘤杀伤活性。此外，我们还表明，在体内异种移植模型中，γ-TCRγδ 比 δ-TCRγδ 表现出更优异的功效。