IL-12、IL-15对急性髓系白血病患者骨髓来源CD34+白血病细胞转化为NK细胞作用研究  

Effects of IL-12 and IL-15 in bone marrow-derived CD34 + leukemia cells from patients with acute myeloid leukemia to differentiate into NK cells

作　　者：李焱[1] 叶敬伟 郭晓玲[2] LI Yan;YE Jingwei;GUO Xiaoling(Department of Hematology,Handan First Hospital,Handan 056002,China)

机构地区：[1]邯郸市第一医院血液内科,河北邯郸056002 [2]河北医科大学第二医院血液内科,050005

出　　处：《临床肿瘤学杂志》2019年第1期32-37,共6页Chinese Clinical Oncology

摘　　要：目的应用细胞因子组合体外诱导CD34+白血病细胞转化为自然杀伤(NK)细胞,探讨白介素(IL)-12、IL-15对NK细胞分化增殖、细胞活性、细胞毒作用以及Granzyme B、TNF-α和IFN-γ基因表达的作用。方法提取26例2016年8月至2017年11月初治高白细胞血症急性髓系白血病患者骨髓单个核细胞(BM-MNC),应用A组(SCF、IL-7、IL-12、IL-15)、B组(SCF、IL-7、IL-2、IL-15)两组细胞因子组合分别体外诱导分化5周,流式细胞术检测NK细胞比例及活化标志,实时定量PCR(QPCR)检测两组NK细胞Granzyme B、TNF-α、IFN-γ等基因的表达水平; CCK-8法分别检测两组NK细胞对患者来源白血病细胞和K562细胞的杀伤率。结果 A组NK细胞比例为(71. 22±2. 25)%,显著高于B组的(56. 32±3. 97)%(P <0. 05)。A组细胞增殖倍数与B组比较,差异无统计学意义(P> 0. 05)。A组NK细胞活化标志CD69+、NKp46+、NKG2D+NK细胞比例分别为(70. 72±3. 62)%、(70. 30±3. 45)%、(72. 75±3. 89)%,均显著高于B组的(59. 98±2. 86)%、(60. 38±2. 84)%和(62. 30±4. 25)%(P <0. 05)。Granzyme B、TNF-α和IFN-γ基因的表达水平较诱导前患者BM-MNC显著升高,A组上述基因表达水平分别为13. 36±1. 54、9. 38±0. 51和6. 26±0. 25,均显著高于B组的9. 82±1. 21、7. 06±0. 52和4. 28±0. 23(P <0. 05)。在效靶比为10∶1时,A组NK细胞对K562细胞和患者来源白血病细胞的杀伤率分别为(62. 20±2. 36)%和(70. 78±2. 54)%,均显著高于B组的(56. 77±3. 81)%和(62. 95±1. 94)%(P <0. 05);且相同效靶比时A、B两组NK细胞对患者来源白血病细胞的杀伤率显著高于K562细胞(P <0. 05)。结论 CD34+白血病细胞可在体外诱导分化为NK细胞,且具有杀伤活性。联合应用IL-12、1L-15细胞因子组合能够诱导分化的NK细胞活性,提高对白血病细胞的杀伤率以及Granzyme B、TNF-α和IFN-γ基因表达水平,且诱导分化的NK细胞对白血病细胞的杀伤具有同体特异性。Objective To apply cytokine combinations to induce CD34+leukemia cells to transform natural killer(NK)cells in vitro,and investigate the effects of interleukin(IL)-12 and IL-15 on NK cell differentiation,cell activity,cytotoxicity,and granzyme B,TNF-αand IFN-γgene expression.Methods Bone marrow mononuclear cells from 26 early treatment patients with acute myeloid leukemia(AML)from August 2016 to November 2017 were extracted.Two groups of cytokine combinations(group A:SCF,IL-7,IL-12 and IL-15;group B:SCF,IL-7,IL-2 and IL-15)were used to induce differentiation for 5 weeks in vitro.The percentage of NK cells and activation markers were measured by flow cytometry.Real-time quantitative PCR(QPCR)was used to detect the expression of Granzyme B,TNF-α,IFN-γof NK cells.The killing rates of NK cells from patients with leukemic cells and K562 cells were measured by CCK-8 assay.Results NK cell ratio in group A was(71.22±2.25)%,significantly higher than(56.32±3.97)%in group B(P<0.05).There was no significant difference between group A and group B in cell proliferation(P >0.05).In group A,CD69^+,NKp46^+,NKG2D^+NK cells were(70.72±3.62)%,(70.30±3.45)%and(72.75±3.89)%,significantly higher than(59.98±2.86)%,(60.38±2.84)%and(62.30±4.25)%of group B(P<0.05).The expression levels of Granzyme B,TNF-αand IFN-γwere significantly higher than those of pre-induced bone marrow mononuclear cells.The expression level of these genes in group A was 13.36±1.54,9.38±0.51 and 6.26±0.25,significantly higher than 9.82±1.21,7.06±0.52 and 4.28±0.23 in group B(P<0.05).In the effective target ratio of 10∶1,the cell killing rates of K562 cells and leukemia cells from patients in group A were(62.20±2.36)%and(70.78±2.54)%,significantly higher than(56.77±3.81)%and(62.95±1.94)%in group B(P<0.05).At the same target ratio,the killing rate of NK cells in leukemia cells derived from patients with group A and B was significantly higher than that of K562 cells(P<0.05).Conclusion CD34^+leukemia cells can induce differentiation into NK cell

关 键 词：急性髓细胞白血病(AML) 白介素-12(IL-12) 白介素-15(IL-15) CD34^+细胞 自然杀伤(NK)细胞 

分 类 号：R733.71[医药卫生—肿瘤]