Full
Outline
To construct CD19-targeted chimeric antigen receptor natural killer (CAR-NK) cells armored with interleukin15 (IL15) or IL15 receptor-linker (RLI) and preliminarily validate their proliferative capacity and anti-tumor activity in vitro.
Natural killer cells (NK92 and NK92MI) from patients with human malignant non-Hodgkin lymphoma were cultured, and CD19-targeted CAR-NK cells armored with IL15 or RLI were prepared using retroviral vector particles. IL15 secretion was measured by ELISA, and proliferative capacity was assessed via CFSE dilution assays. Human B-lymphocytic leukemia cells (Nalm6-GFP-Luc) and human colon cancer cells overexpressing CD19 (hCD19-SW620-GFP-Luc) were cultured, with surface CD19 expression confirmed (>99% positivity for both). Anti-tumor activity was evaluated by measuring cytotoxicity at effector-to-target (E:T) ratios using luciferase-based assays (4/12 h), detecting surface CD107a expression via flow cytometry, and quantifying cytokine release using CBA assays.
NK92/NK92MI-CD19 CAR cells armored with IL15 or RLI were successfully generated. IL15 secretion was significantly higher in armored groups versus non-armored controls (P<0.01). Without IL-2 stimulation, IL15/RLI enhanced CAR-NK proliferation (P<0.05). Both armored designs significantly increased tumor-killing efficiency (P<0.05) and CD107a degranulation. IL15/RLI-armored NK92-CD19 CAR cells exhibited elevated release of IL2, IL10, IL6, TNF-α, sFas, IFN-γ, and Granulysin (P<0.05), while armored NK92MI-CD19 CAR cells showed additional increases in Granzyme A, Granzyme B, and Perforin (P<0.05).
IL15/RLI-armored NK92/NK92MI-CD19 CAR cells demonstrate potent anti-tumor activity, supporting their combined clinical therapeutic potential for tumors.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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