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目的探讨体外高效扩增多发性骨髓瘤(MM)患者自身自然杀伤细胞(NK细胞)的方法,研究扩增前和扩增后NK细胞对骨髓瘤细胞株U266和自体骨髓瘤细胞的杀伤作用。方法 10例MM患者的外周血单个核细胞与K562转基因细胞株在含不同浓度白细胞介素(IL)-2的培养液中共育14d,应用流式细胞仪,采用氚标记的胸腺嘧啶核苷(3H-TdR)掺入法和铬51释放法等方法,研究NK细胞的扩增情况、增殖能力、免疫表型和抗骨髓瘤作用。结果 300U/mLIL-2共培养体系扩增NK细胞的扩增倍数为196.4±12.8,扩增后NK细胞存活率>95%,IL-2能增强共培养体系中NK细胞的增殖能力。在效靶比为10∶1时,扩增后NK细胞对自体骨髓瘤细胞的平均杀伤率为(38.1±6.2)%,显著高于扩增前的(3.9±1.7)%(P<0.01)。结论 K562转基因细胞株能特异性刺激骨髓瘤患者NK细胞扩增,扩增后NK细胞抗自体骨髓瘤的作用显著增强,扩增并活化的NK细胞将有望作为临床治疗MM的免疫治疗方法。
OBJECTIVE: To explore a method for efficient expansion of natural killer cells (NK cells) in patients with multiple myeloma (MM) in vitro and to study the killing effect of NK cells on myeloma cell line U266 and autologous myeloma cells before and after amplification. Methods Peripheral blood mononuclear cells from 10 MM patients were co-cultured with K562 transgenic cell line in culture medium containing different concentrations of interleukin (IL) -2 for 14 days. Flow cytometry was used to detect the expression of thymidine 3H-TdR) incorporation method and Chromium 51 release method to study the expansion of NK cells, proliferation, immunophenotype and anti-myeloma. Results The multiplication of NK cells expanded by 300U / mL IL-2 co-culture system was 196.4 ± 12.8, and the survival rate of NK cells was> 95% after expansion. IL-2 could enhance the proliferation of NK cells in co-culture system. When the target ratio was 10: 1, the average killing rate of NK cells to autologous myeloma cells was (38.1 ± 6.2)%, significantly higher than that before expansion (3.9 ± 1.7%) (P <0.01) . Conclusion The K562 transgenic cell line can specifically stimulate the expansion of NK cells in patients with myeloma. After NK cell expansion, the effect of NK cells on autoantibody myeloma is significantly enhanced. Expanded and activated NK cells are expected to be used as immunotherapy for MM.