骨髓增生异常综合征患者单个核细胞诱导的巨噬细胞的特征及功能

Characteristic and function of peripheral blood mononuclear cells-induced macrophages in patients with myelodysplastic syndrome

目的观察骨髓增生异常综合征（MDS）患者单个核细胞诱导的巨噬细胞的功能，分析其与MDS疾病发展的关系。方法以2014年9月至2015年12月天津医科大学总医院血液科24例MDS患者为病例组，以15名健康志愿者为正常对照组。收集外周血标本并分离单个核细胞，体外以GM—CSF诱导分化形成巨噬细胞。在光学显微镜下观察巨噬细胞的形态；使用流式细胞术检测巨噬细胞的数量及其表面受体CD206、SIRPot的表达水平；分别采用流式细胞术和免疫荧光显微镜检测巨噬细胞的吞噬功能。结果MDS来源的巨噬细胞形态较正常对照差（形态不规则、细胞体积小、异形明显、无足突、数量减少），MDS外周血单个核细胞转化为巨噬细胞的转化率为（5．17±3．47）％，显著低于正常对照组的（66．18±13．43）％（t=3．529，P=0．001）；MDS来源的巨噬细胞表面识别受体CD206的表达水平[（9．73±2．59）％]显著低于正常对照组[（51．15±10．82）％]（t=-4．551，P〈0．001），SIRPa的表达水平[（0．51±0．09）％]显著低于正常对照组[（O．77±0．06）％]（t=2．102，P=-0．043）；@MDS来源的巨噬细胞吞噬免疫微球的吞噬指数、吞噬百分率分别为0．45±0．08、（23．69±3．22）％，显著低于正常对照组的0．92±0．07、（42．75±2．13）％（P值均〈0．05），MDS来源的巨噬细胞吞噬单个核细胞的吞噬指数（0．24±0．04）显著低于正常对照组（0．48±0．96，t=-3．464，P=0．001）。结论MDS患者外周血单个核细胞诱导形成巨噬细胞能力下降，吞噬识别受体减少，吞噬能力减弱。

Objective To explore characteristic and function of peripheral blood mononuclear cells （PBMNC）-induced macrophages in patients with myelodysplastic syndrome （MDS） to couple with its progression. Methods A total of 24 MDS patients （ll low-risk patients and 13 high-risk group patients） referred to Department of Hematology of Tianjin Medical University General Hospital and normal controls were enrolled from September 2014 to December 2015. PBMNC was stimulated with GM- CSF to transform to macrophages. The morphology of macrophages was observed by microscope. The quantity of macrophages, CD206 and SIRPct on surface of macrophages were detected by flow cytometry. The phagocytic function of macrophages was analyzed by fluorescence microscopy and flow cytometry. Results The morphology of macrophages from MDS patients was abnormal. The percentage of transformed macrophages was （5.17±3.47）% in patients with MDS, which was lower than that in controlssignificantly [ （66.18± 13.43）%, t=3.529, P=0.001 ]. The expression of CD206 on macrophages from MDS patients was significantly lower than that of controls [ （9.73±2.59）% vs （51.15± 10.82）%, t=4.551, P〈 0.001 ]. The SIRPa level of rnacrophages from MDS patients was significantly lower than that of controls [ （0.51 ±0.09）% vs （0.77±0.06）%, t=2.102, P=0.043]. The phagocytic index and the percentage of phagocytic of macrophages from MDS patients were significantly lower than those of macrophages from normal controls [ 0.45±0.08 vs 0.92±0.07, t=-6.253, P=0.008; （23.69±3.22） % vs （42.75±2.13） %, t=- 6.982, P=0.006 respectively] by flow cytometry. The phagocytic index of MDS patients was significantly lower than that of controls （0.24±0.04 vs 0.48±0.96, t=3.464, P=0.001） by fluorescence microscopy. Conclusion The quantity, recognization receptors and phagocytosis of PBMNC- induced macrophages decreased in MDS patients.