Chen Wenxiu, Tian Manman, Shen Jie, Liu Qin, Li Rutian, Liu Baorui, Shao Jie
Comprehensive Cancer Centre of Drum Tower Hospital, Medical School of Nanjing University, Clinical Cancer Institute of Nanjing University, Nanjing, China.
Comprehensive Cancer Centre of Drum Tower Hospital, Medical School of Nanjing University, Clinical Cancer Institute of Nanjing University, Nanjing, China.
Int Immunopharmacol. 2025 Dec 3;166:115569. doi: 10.1016/j.intimp.2025.115569. Epub 2025 Sep 22.
Adoptive transfer of NK or T cells alone requires a long preparation time. We developed a rapid co-culture system, including cytokine-induced memory-like NK (CIML NK) cells and activated T cells (Ac-T), as well as tumor-penetrating peptide iRGD.
PBMCs were pretreated with IL-12/15/18 for 16 h, then washed to induce a memory NK phenotype. K562-CD48-41BBL-mbIL-21 cells were generated by lentiviral transduction and used as feeder cells to expand and activate NK cells from PBMCs for 7 days. On day 7, an anti-CD3 monoclonal antibody (OKT3) and an anti-CD28 monoclonal antibody (CD28.2) were added and washed overnight for 24 h. The in vitro (cytotoxicity, cytokines) and in vivo (tumor inhibition, survival) functions of iRGD-modified CIML NK&Ac-T cells (CIML NK&Ac-T-iRGD) were compared to CIML NK&T-iRGD without addition of CD3/CD28.
We established a one-week rapid co-culture system comprising NK and T cells. This system markedly increased cytokine secretion and demonstrated potent in vitro cytotoxicity against gastric cancer cell lines, significantly inhibited tumor growth, and prolonged survival in a gastric cancer xenograft model. Notably, iRGD-modified CIML NK&Ac-T cells showed superior efficacy compared with CIML NK&T cells.
Our data suggest that the rapid co-culture system of iRGD-modified CIML NK&Ac-T cells used for adoptive cell transfer demonstrates potent anti-tumor effects in gastric cancer. This approach is time-efficient and cost-saving, with potential for broader clinical application compared with conventional NK cell therapies.
单独进行自然杀伤（NK）细胞或T细胞的过继性转移需要较长的准备时间。我们开发了一种快速共培养系统，包括细胞因子诱导的记忆样NK（CIML NK）细胞和活化T细胞（Ac-T），以及肿瘤穿透肽iRGD。
将外周血单个核细胞（PBMC）用白细胞介素（IL）-12/15/18预处理16小时，然后洗涤以诱导记忆NK表型。通过慢病毒转导产生K562-CD48-41BBL-mbIL-21细胞，并用作饲养细胞，从PBMC中扩增和激活NK细胞7天。在第7天，加入抗CD3单克隆抗体（OKT3）和抗CD28单克隆抗体（CD28.2），并过夜洗涤24小时。将iRGD修饰的CIML NK&Ac-T细胞（CIML NK&Ac-T-iRGD）的体外（细胞毒性、细胞因子）和体内（肿瘤抑制、生存）功能与未添加CD3/CD28的CIML NK&T-iRGD进行比较。
我们建立了一个包含NK细胞和T细胞的一周快速共培养系统。该系统显著增加了细胞因子分泌，并对胃癌细胞系表现出强大的体外细胞毒性，在胃癌异种移植模型中显著抑制肿瘤生长并延长生存期。值得注意的是，iRGD修饰的CIML NK&Ac-T细胞比CIML NK&T细胞显示出更好的疗效。
我们的数据表明，用于过继性细胞转移的iRGD修饰的CIML NK&Ac-T细胞快速共培养系统在胃癌中显示出强大的抗肿瘤作用。这种方法省时且节省成本，与传统NK细胞疗法相比具有更广泛的临床应用潜力。
