A novel feeder cell based on 4-1BBL and membrane-bound IL-21/IL-15 induce highly expansion and anti-tumor effect of natural killer cells

作者：Sha Gong, Nan Mei, Jun Wang, Junsheng Zhu, Lu Wang, Xiaohong Lü, Pengcheng He, Weiwei Chen, Lei Xi, Yingying Bao, David N. Wald, Xiaohu Fan, Huaiyu Wang · 发表于：BMC Biotechnology · 年份：2025 · DOI：10.1186/s12896-025-01024-x · 被引用次数：4 · 研究领域：Immune Cell Function and Interaction、CAR-T cell therapy research、Cancer Research and Treatments

BACKGROUND: Natural killer (NK) cell immunotherapy is a promising approach for cancer treatment. However, its extensive clinical application was limited to the large-scale clinical-grade expansion of NK cells. In this study, we expanded NK cells from healthy donor's peripheral blood mononuclear cells (PBMCs) using a newly designed K562 feeder cell line. METHODS: The feeder cells were generated by transducing K562 cells with lentiviral particles carrying 4-1BBL and mbIL-21/-15. NK cells were expanded from PBMCs with these genetically modified, frozen-thawed and irradiated K562 feeder cells in the presence of IL-2. The purity, quantity, and receptors expression of the expanding NK cells were dynamically monitored. Furthermore, their anti-tumor efficacy was evaluated both in vitro and in vivo following a two-week expansion period. RESULTS: The K562-4-1BBL-mbIL-21/-15 feeder cells induced highly-efficient NK cells expansion from PBMCs (17902-fold) within two weeks. There was a notable upregulation in the expression of activating receptors including NKG2D, NKp30, NKp44, and NKp46 during the expansion process. Moreover, the expanded NK cells displayed enhanced cytotoxicity against a variety of hematological (K562, MOLM-13, OCI-AML-3, THP-1) and solid (Hep-G2, OVCAR3) cancer cell lines in vitro. In the humanized U937 xenograft mouse model, the NK cells extended the median survival time of the AML-bearing mice from 19.40 to 28.25 days. CONCLUSIONS: We have successfully established a ...