Luangwattananun Piriya, Sangsuwannukul Thanich, Supimon Kamonlapat, Thuwajit Chanitra, Chieochansin Thaweesak, Sa-Nguanraksa Doonyapat, Samarnthai Norasate, O-Charoenrat Pornchai, Junking Mutita, Yenchitsomanus Pa-Thai
Siriraj Center of Research Excellence for Cancer Immunotherapy (SiCORE-CIT), Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand; Division of Molecular Medicine, Research Department, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Siriraj Center of Research Excellence for Cancer Immunotherapy (SiCORE-CIT), Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand; Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Int Immunopharmacol. 2023 Nov;124(Pt B):111012. doi: 10.1016/j.intimp.2023.111012. Epub 2023 Oct 5.
T cell-based immunotherapy has transformed cancer treatment. Nonetheless, T cell antitumor activity can be inhibited by an immune checkpoint molecule expressed on cancer cells, program death ligand 1 (PD-L1), which interacts with the PD-1 on T cells. We generated αPD-L1 × αCD3 bispecific T-cell engager-armed T cells (BATs) to prevent PD-L1/PD-1 interaction and hence to redirect T cells to kill cancer cells. αPD-L1 × αCD3 bispecific T-cell engagers (BTEs) were produced from Chinese hamster ovary (CHO) cells to arm human primary T cells. Flow cytometry was used to investigate BTE binding to BATs. The cytotoxicity of BATs against PD-L1-expressing breast cancer (BC) cell lines was assessed in 2-dimensional (2D) and 3-dimensional (3D) culture models. The binding stability of BTE on BATs and their efficacy after cryopreservation were also examined. The CHO cell BTE expression yield was 3.34 mg/ml. The binding ability on T cells reached 91.02 ± 4.2 %. BATs specifically lysed PD-L1-expressing BC cells, with 56.4 ± 15.3 % HCC70 cells and 70.67 ± 15.6 % MDA-MB-231 cells lysed at a 10:1 effector-to-target ratio. BATs showed slight, nonsignificant lysis of PD-L1-negative BC cells, MCF-7, and T47D. Moreover, BATs significantly disrupted MDA-MB-231 3D spheroids expressing PD-L1 after 48 and 72 h of coculture. Cryopreserved BATs maintained BTE binding stability, cell viability, and anticancer activity, comparable to fresh BATs. αPD-L1 × αCD3 BATs induced the cytolysis of PD-L1-expressing BC cells in 2D and 3D coculture assays. BATs can be prepared and preserved, facilitating their use and transportation. This study demonstrates the potential of αPD-L1 × αCD3 BATs in treating cancers with positive PD-L1 expression.
基于T细胞的免疫疗法已经改变了癌症治疗。尽管如此，T细胞的抗肿瘤活性可被癌细胞上表达的一种免疫检查点分子程序性死亡配体1（PD-L1）所抑制，该分子与T细胞上的程序性死亡受体1（PD-1）相互作用。我们制备了αPD-L1×αCD3双特异性T细胞衔接器武装的T细胞（BATs），以阻止PD-L1/PD-1相互作用，从而使T细胞重新定向以杀伤癌细胞。αPD-L1×αCD3双特异性T细胞衔接器（BTEs）由中国仓鼠卵巢（CHO）细胞产生，用于武装人原代T细胞。采用流式细胞术研究BTE与BATs的结合情况。在二维（2D）和三维（3D）培养模型中评估BATs对表达PD-L1的乳腺癌（BC）细胞系的细胞毒性。还检测了BTE在BATs上的结合稳定性及其冻存后的疗效。CHO细胞BTE的表达产量为3.34mg/ml。其对T细胞的结合能力达到91.02±4.2%。BATs特异性裂解表达PD-L1的BC细胞，在效应细胞与靶细胞比例为10:1时，HCC70细胞的裂解率为56.4±15.3%，MDA-MB-231细胞的裂解率为70.67±15.6%。BATs对PD-L1阴性的BC细胞MCF-7和T47D表现出轻微的、无显著意义的裂解。此外，在共培养48小时和72小时后，BATs显著破坏了表达PD-L1的MDA-MB-231三维球体。冻存后的BATs保持了BTE的结合稳定性、细胞活力和抗癌活性，与新鲜的BATs相当。αPD-L1×αCD3 BATs在二维和三维共培养试验中诱导表达PD-L1的BC细胞发生细胞溶解。BATs可以制备和保存，便于其使用和运输。本研究证明了αPD-L1×αCD3 BATs在治疗PD-L1表达阳性癌症中的潜力。
