目的：探讨人源IL-15转基因NCG小鼠（NCG-hIL-15鼠）在CAR-NK细胞肿瘤治疗临床前评价中的作用。方法：通过qPCR结合WB法检测NCG-hIL-15鼠骨髓及主要器官（脾、肝、肺、肾和胰）中的表达丰度。将人 PBMC 来源的NK（PB-NK）细胞回输到NCG-hIL-15鼠及对照NCG鼠，监测NK细胞的体内扩增与受体鼠的体质量、生存期变化，流式细胞术检测体内扩增NK细胞的活化受体与抑制受体表达差异 ，WB 法检测穿孔素与颗粒酶 B 的表达。MIAPaca-2 细胞原位荷瘤NCG-hIL-15鼠或NCG鼠用anti-MUC1-CAR-NK细胞回输治疗，流式细胞术检测CAR-NK细胞在不同组鼠体内的存活情况，小动物活体成像术检测肿瘤生长并记录荷瘤鼠生存期。结果：NCG-hIL-15鼠中PB-NK细胞可以在10周内稳定增殖，且观察期内未发生显著的移植物抗宿主反应。NCG-hIL-15鼠中扩增NK细胞的KAR与KIR丰度与原始细胞相比差异较小；与NCG鼠体内增殖的NK细胞相比，NCG-hIL-15鼠体内扩增的NK细胞有更高水平的穿孔素与颗粒酶B表达（均P<0.05）。CAR-NK细胞在NCG-hIL-15鼠体内存活比显著增加、抑瘤效果增强，与对照组相比NCG-hIL-15鼠生存期显著延长（均P<0.01）。结论：NCG-hIL-15鼠模型可用于基于NK细胞免疫疗法的临床前实验与生物学评价。
Objective: To explore the application value of human IL-15 transgenic NCG mice (NCG-hIL-15 mice) in preclinical evaluation of chimeric antigen receptor modified NK (CAR-NK) cell therapy for tumor treatment. Methods: qPCR and WB were performed to detect the expression of human IL-15 in the bone marrow and main organs (spleen, liver, lung, kidney and pancreas) of transgenic mice. After being transfused with human PBMC-derived NK (PB-NK) cells, the NCG-hIL-15 mice and control NCG mice were continuously monitored for the in vivo amplification of NK cells and the changes in body weight and survival time. Flow cytometry was used to detect the differential expressions of activated receptors and inhibitory receptors in amplified NK cells. WB was used to detect the expressions of perforin and granzyme-B. NCG-hIL-15 mice or NCG mice bearing MIAPaca-2 cell transplanted tumor were treated with anti-MUC1-CAR-NK cell reinfusion; then, the CAR-NK cell survival in different groups of mice was detected by Flow cytometry, and the survival time of tumor bearing mice was recorded and tumor growth was detected by in vivo imaging. Results:The results indicated that PB-NK cells could proliferate stably within 10 weeks in NCG-hIL-15 mice without obvious graft versus host diseases (GVHD) during the observation period. The in vivo-expanded human NK cells maintained the original expression patterns of various surface molecules, including KARs and KIRs. Compared with the NK cells in NCG mice, the NK cells in NCG-hIL-15 mice contained significantly higher amounts of granzyme-B and perforin (all P<0.05). CAR-NK cells showed significantly increased survival rate and stronger tumor-inhibitory effect in NCG-hIL-15 mice as compared with those in control NCG mice, resulting in significantly prolonged survival in NCG-hIL-15 mice (all P<0.01). Conclusion: NCG-hIL-15 mouse model has potential application value in preclinical trial and biological evaluation of NK cell-based immunotherapy.