依鲁替尼对多发性骨髓瘤Treg细胞的影响及相关作用机制  

Effects of Ibrutinib on Treg cells in multiple myeloma and related mechanisms

作　　者：李胜利[1] 姜杨 王琰 徐源[1] 张洋 杨水个 马金凤[1] 孙道萍[1] 孙忠亮[1] LI Sheng-li;JIANG Yang;WANG Yan(Department of Hematology,Jining NO.1 People's Hospital,Jining Shandong 272100,China;Department of Hematology,The Second Hospital of Shandong University,Ji'nan Shandong 250033,China)

机构地区：[1]济宁市第一人民医院血液内科,山东济宁272100 [2]山东大学第二医院血液内科,山东济南250033

出　　处：《临床和实验医学杂志》2020年第17期1800-1804,共5页Journal of Clinical and Experimental Medicine

基　　金：山东省医药卫生科技发展计划项目(编号:2017WS656);济宁市重点研发科技项目(编号:2018SMNS008)。

摘　　要：目的探究依鲁替尼对多发性骨髓瘤Treg细胞表达水平的影响及其作用机制。方法选取2018年6~12月济宁市第一人民医院收治的10例多发性骨髓瘤患者作为研究对象,同时选取同期来院的10例无关疾病(白细胞减少症或缺铁性贫血)患者作为对照组。分别采集两组患者的骨髓液,采用酶联免疫吸附测定(ELISA)检测骨髓浆液中可溶性程序性死亡配体1(PD-L1)表达。流式细胞术测定RPMI-8226细胞PD-L1表达变化。选取18只6~8周龄SPF级C57BL/6雄性小鼠,构建骨髓瘤小鼠模型,将骨髓瘤小鼠按随机分为3组:对照组、依鲁替尼组和依鲁替尼联合IL-6组,每组各6只,对照组小鼠给予二甲基亚砜皮下注射8 d,依鲁替尼组小鼠给予6 mg/kg依鲁替尼皮下注射8 d,依鲁替尼联合IL-6组小鼠接受6 mg/kg依鲁替尼联合1 ng/kg IL-6皮下注射8 d。采用流式细胞术检测各组实验小鼠在依鲁替尼作用前后Treg细胞变化情况,以DC4+CD25highT细胞占CD4+细胞的百分比代表Treg细胞水平。结果与对照组患者相比,多发性骨髓瘤患者骨髓液中可溶性PD-L1表达明显增高,差异具有统计学意义(P <0.01)。与对照组RPMI-8226细胞相比,依鲁替尼组RPMI-8226细胞PD-L1呈现明显低表达,依鲁替尼联合IL-6组RPMI-8226细胞PD-L1表达水平较依鲁替尼组则明显增高,与对照组RPMI-8226细胞相比则明显降低,差异均具有统计学意义(P <0.01)。与对照组小鼠相比,依鲁替尼组小鼠骨髓液Treg细胞数明显减少,依鲁替尼联合IL-6组小鼠骨髓液Treg细胞数明显低于对照组,同时明显高于依鲁替尼组,差异均具有统计学意义(P <0.05);与对照组小鼠相比,依鲁替尼组小鼠骨髓液中可溶性PD-L1呈现明显低表达,依鲁替尼联合IL-6组小鼠PD-L1表达水平较依鲁替尼组小鼠则明显增高,与对照组小鼠相比则明显降低,差异均具有统计学意义(P <0.05)。结论依鲁替尼可通过介导多发性骨髓瘤中Treg细胞占比变化Objective To examine the impact of Ibrutinib on expression of Treg cells in multiple myeloma and its mechanisms.MethodsThe study included ten patients who were newly diagnosed with multiple myeloma between June 1 and December 31 in 2018 in the First People’s Hospital of Jining.The control group contains 10 cases of patients (Leukopenia or Iron-deficiency anemia) without hematological tumor enrolled at the same period.The level of Soluble PD-1ligand (PD-L1) of bone marrow from the two groups of patients were detected by the enzyme-linked immunosorbent assay.The change of PD-L1 expression in RPMI8226 (R8) were evaluated using flow cytometry method.A total of 18 SPF C57BL/6 male mice of 6~8 weeks was used to construct myeloma mice model.The myeloma-implanted mice were randomly divided into 3 groups with 6 mice in each group:control group,Ibrutinib and Ibrutinib+IL-6 group.The control mice were treated with subcutaneous injection of DMSO;for Ibrutinib group mice,6 mg/kg Ibrutinib were subcutaneously injected;Ibrutinib+IL-6 groups of mice accepted 6 mg/kg Ibrutinib and 1ng/kg IL-6 through subcutaneous injection;all the mice were treated for 8 days.Flow cytometry was used to detect the change of Treg cells before and after the treatment of Ibrutinib in the experimental mice.The percentage of CD4+ CD25highT cells in CD4+ cells were accounting for the Treg cells.Results Compared with the control group,the expression of soluble PD-L1 in the bone marrow fluid of myeloma patients was significantly higher,the difference was statistically significant (P<0.01).Compared with the control group,the expression of PD-L1 in RPMI-8226 cells in Ibrutinib group was significantly decreased;for Ibrutinib+IL-6 group,the expression of PD-L1 was significantly lower than that in the control group,but significantly higher than that in the Ibrutinib group,the differences were statistically significant (P<0.01).Compared with the control mice group,the number of Treg cells in Ibrutinib group was significantly decreased,the number of Treg cells

关 键 词：多发性骨髓瘤 患者 小鼠 依鲁替尼 TREG细胞 免疫微环境 

分 类 号：R733.3[医药卫生—肿瘤]