Zhao Yueqi, Li Yinghui, Wang Shuaiqi, Han Jingyi, Lu Mingyang, Xu Yupeng, Qiao Wenhua, Cai Menghua, Xu Yi, Hu Yu, Zhang Jianmin, Chen Hui, He Wei
Department of Immunology, CAMS Key Laboratory T-Cell and Cancer Immunotherapy, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, State Key Laboratory of Common Mechanism Research for Major Diseases, Beijing 100005, China.
Department of Thoracic Surgery, Qilu Hospital of Shandong University, Jinan 250012, China.
Cancers (Basel). 2025 Mar 17;17(6):998. doi: 10.3390/cancers17060998.
Claudin18.2 (CLDN18.2) is highly expressed during the development of various malignant tumors, especially gastric cancer, and CAR-T cells targeting CLDN18.2 have therapeutic potential. However, their dependence on the major histocompatibility complex (MHC) for antigen recognition limits their application. Human Gamma Delta (γδ) T cells, with strong MHC-independent cytotoxicity to most solid tumors both in vivo and in vitro, are emerging as ideal cells for the generation of robust universal CLDN18.2 CAR-T cells to treat solid tumors. Our aim was to construct a universal CAR-γδ T cell targeting CLDN18.2.
We constructed novel CAR-CLDN18.2-γδ T cells by lentiviral infection and compared their superior efficacy in the treatment of CLDN18.2-positive solid tumors in vivo and in vitro.
CD3ζ expression was verified in HEK293T cells after lentiviral transfection of CLDN18.2 CAR, and the lentivirus was packaged and concentrated to a titer of 4.90 × 10 TU/mL. Primary γδ T cells and αβ T cells were infected with efficiencies of approximately 31.76 ± 4.122% and 44.13 ± 4.436%, respectively. CAR-CLDN18.2-γδ T cells exhibited specific cytotoxicity against CLDN18.2-positive gastric cancer cells and secreted relatively high levels of Granzyme-B, Perforin-1, and IFN-γ. CAR-γδ T cells also showed superior cytotoxicity to target cells compared to classical CAR-αβ T cells in vitro. Finally, the antitumor activity of γδ T-CAR-CLDN18.2 cells was evaluated in tumor-bearing NSG mice, and CAR-CLDN18.2-γδ T cells significantly inhibited tumor growth and prolonged the survival of the mice.
Our results demonstrate that universal CAR-CLDN18.2-γδ T cell is promising for the treatment of CLDN18.2-positive solid tumor and provide insights for the development of more universal CAR-γδ T-cell strategies for tumor immunotherapy.
紧密连接蛋白18.2（CLDN18.2）在多种恶性肿瘤尤其是胃癌的发生发展过程中高表达，靶向CLDN18.2的嵌合抗原受体T细胞（CAR-T细胞）具有治疗潜力。然而，它们对主要组织相容性复合体（MHC）进行抗原识别的依赖性限制了其应用。人γδ T细胞在体内和体外对大多数实体瘤均具有强大的不依赖MHC的细胞毒性，正逐渐成为生成强大的通用型CLDN18.2 CAR-T细胞以治疗实体瘤的理想细胞。我们的目的是构建一种靶向CLDN18.2的通用型CAR-γδ T细胞。
我们通过慢病毒感染构建新型CAR-CLDN18.2-γδ T细胞，并比较其在体内和体外治疗CLDN18.2阳性实体瘤方面的卓越疗效。
慢病毒转染CLDN18.2 CAR后，在HEK293T细胞中验证了CD3ζ的表达，且慢病毒包装浓缩后的滴度为4.90×10 TU/mL。原代γδ T细胞和αβ T细胞的感染效率分别约为31.76±4.122%和44.13±4.436%。CAR-CLDN18.2-γδ T细胞对CLDN18.2阳性胃癌细胞表现出特异性细胞毒性，并分泌相对高水平的颗粒酶B、穿孔素-1和干扰素-γ。与经典的CAR-αβ T细胞相比，CAR-γδ T细胞在体外对靶细胞也表现出更强的细胞毒性。最后，在荷瘤NSG小鼠中评估了γδ T-CAR-CLDN18.2细胞的抗肿瘤活性，CAR-CLDN18.2-γδ T细胞显著抑制肿瘤生长并延长了小鼠的生存期。
我们的结果表明，通用型CAR-CLDN18.2-γδ T细胞在治疗CLDN18.2阳性实体瘤方面具有前景，并为开发更多用于肿瘤免疫治疗的通用型CAR-γδ T细胞策略提供了思路。
